Natural Polymers: Volume 1: Composites

Natural Polymers
Volume 1: Composites
RSC Green Chemistry
Series Editors:
James H Clark, Department of Chemistry, University of York, UK
George A Kraus, Department of Chemistry, Iowa State University, Ames, Iowa,
USA
Andrzej Stankiewicz, Delft University of Technology, The Netherlands
Peter Siedl, Federal University of Rio de Janeiro, Brazil
Yuan Kou, Peking University, People’s Republic of China
Titles in the Series:

1: The Future of Glycerol: New Uses of a Versatile Raw Material
2: Alternative Solvents for Green Chemistry
3: Eco-Friendly Synthesis of Fine Chemicals
4: Sustainable Solutions for Modern Economies
5: Chemical Reactions and Processes under Flow Conditions
6: Radical Reactions in Aqueous Media
7: Aqueous Microwave Chemistry
8: The Future of Glycerol: 2nd Edition
9: Transportation Biofuels: Novel Pathways for the Production of Ethanol,
Biogas and Biodiesel
10: Alternatives to Conventional Food Processing
11: Green Trends in Insect Control
12: A Handbook of Applied Biopolymer Technology: Synthesis, Degradation
and Applications
13: Challenges in Green Analytical Chemistry
14: Advanced Oil Crop Biorefineries
15: Enantioselective Homogeneous Supported Catalysis
16: Natural Polymers Volume 1: Composites
17: Natural Polymers Volume 2: Nanocomposites
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Natural Polymers
Edited by
Maya J John
CSIR Materials Science and Manufacturing, Port Elizabeth, South Africa and
Department of Textile Science, Faculty of Science, Nelson Mandela Metropolitan
University, Port Elizabeth, South Africa
Thomas Sabu
School of Chemical Sciences, Mahatma Gandhi University, Kottayam, India
RSC Green Chemistry No. 16
ISBN: 978-1-84973-402-8
ISSN: 1757-7039
A catalogue record for this book is available from the British Library
r The Royal Society of Chemistry 2012
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Preface
Natural polymers offer an alternative solution to the growing environmental

threat and looming petroleum crisis. The use of natural polymers for many
applications would therefore contribute to creating a sustainable economy.
In contrast, the feedstocks for polymers derived from petrochemicals will in
the long run eventually perish. Polymer chemists, physicists and engineers
show great interest in the development of eco-friendly micro- and nano-
structured functional materials based on natural polymers. In recent years,
natural polymers have generated much interest due to their unique morphol-
ogy and physical properties. The growing interest among academics and
industrial researchers in the field of natural polymers is the driving force
behind the present book.
The book is divided into two volumes: the first covers natural polymer
composites and the second deals with natural polymer nanocomposites.
Volume 1 comprises two sections reviewing (1) natural fibres and composites
and (2) protein fibres and composites. Under natural fibres and composites, the
characterization and new sources of natural fibres are discussed. It also looks
into whether natural fibres can indeed be a replacement for synthetic fibres
in industrial applications. Under protein fibres and composites, important
advancements in the field of silk, spider silk and mussel fibres are discussed.
Volume 2 deals with the properties and characterization of cellulose, chitosan,
furanic, starch and silk nanocomposites. A final chapter touches upon the
industrial applications of natural polymer nanocomposites.
This book is unique in the sense that it deals exclusively with some of the
important polymers found in nature, modifications of natural polymers and
tailoring them into composites and nanocomposites. In addition, it covers
novel topics related to the properties and characterization of mussel fibres and
sea shells.

Natural Polymers, Volume 1: Composites
Edited by Maya J John and Thomas Sabu
Published by the Royal Society of Chemistry, www.rsc.org
v
vi Preface
As the editors of this book, we have enjoyed working with the individual
contributors from different parts of the world and appreciate their diligence
and patience. We would also like to thank all the publishers who generously
gave their permission to reprint material.
Maya Jacob John and Sabu Thomas

Contents
Chapter 1 Natural Polymers: An Overview 1
Maya Jacob John and Sabu Thomas
1.1 Introduction 1
1.2 Natural Polymer Research 4
1.2.1 Natural Fibres 4
1.2.2 Protein Fibres 6
References 7
Chapter 2 Biomimetics: Inspiration from the Structural Organization

of Biological Systems 8
Kalpana S. Katti, Chunju Gu and Dinesh R. Katti
2.1 Introduction: Hierarchy and Structural Order 8

2.2 Biological Materials Systems 8
2.2.1 Bone 8
2.2.2 Teeth 14
2.2.3 Seashells 18
2.2.4 Spider Silk 24
2.3 Biomimetic Design and Fabrication 29
2.4 Perspectives on Future of Biomimetic Technologies 30
References 31

vii
viii Contents
Chapter 3 Natural Fibres as Composite Reinforcement Materials:
Description and New Sources 37
Karine Charlet
3.1 Introduction 37
3.2 Advantages and Drawbacks of Natural Fibres as
Composite Reinforcement Materials 38
3.3 Description of New Sources of Vegetable Fibres 48
3.4 Concluding Remarks 56
References 57
Chapter 4 Relation between Structural Anisotropy in Natural Fibres

and Mechanical Properties in Composites 63
Elessandra da Rosa Zavareze and
Alvaro Renato Guerra Dias
4.1 Introduction 63
4.2 Natural Fibres 65
4.3 Sources of Natural Fibres 66
4.3.1 Cotton Fibre 67
4.3.2 Jute Fibre 67
4.3.3 Flax Fibre 67
4.3.4 Ramie Fibre 67
4.3.5 Hemp Fibre 67
4.3.6 Sisal Fibre 68
4.3.7 Henequen Fibre 68
4.3.8 Coir Fibre 68
4.4 Composites 69
4.4.1 Biopolymer Composites 69
4.4.2 Thermoplastic Composites 70
4.4.3 Natural Fibre Composites 71
4.4.4 Anisotropy 73
4.4.5 Mechanical Properties 75
4.4.6 Effect of Anisotropy in Natural Fibres on
Mechanical Properties 77
4.5 Applications of Natural Fibre Composites 80
4.6 Final Considerations 81
References 82
Chapter 5 Flame Retardant Characteristics of Natural Fibre

Composites 86
Baljinder K. Kandola
5.1 Introduction 86
5.2 Thermal Stability and Flammability of Natural Fibres 88
Contents ix
5.3 Flammability of Composite Matrices 92
5.3.1 Thermoplastics 92
5.3.2 Biodegradable Matrices 94
5.3.3 Natural Rubber 96
5.3.4 Thermosets 97
5.4 Flammability of Composites 99
5.4.1 Performance Requirements Depending Upon
Application Areas 99
5.4.2 Key Parameters Affecting the Flammability of
Composites 100
5.4.3 Flammability Data 101
5.5 Flame Retardant Strategies for Components/
Composites 106
5.5.1 Flame Retardant Matrices: Use of Reactive or
Additive Flame Retardants 106
5.5.2 Treatment of Natural Fibres with Flame
Retardants 110
5.5.3 Surface Treatments of Composites 112
5.6 Summary 113
References 113
Chapter 6 Natural Fibre Composites: Automotive Applications 118

S. C. R. Furtado, A. J. Silva, C. Alves, Luı´s Reis,
Manuel Freitas and Paulo Ferrão
6.1 Introduction 118

6.2 Composite Materials: Definition and Classification 121
6.2.1 Matrices 122
6.2.2 Reinforcements: Fibres and Fillers 122
6.3 Natural Fibres 124
6.4 Vegetable Fibres: Advantages and Disadvantages 126
6.4.1 Production 126
6.4.2 Environment/Health 127
6.4.4 Physical and Chemical Properties 128
6.4.5 Processing 128
6.5 Applications in the Automotive Industry 129
6.6 Future Perspectives 134
6.7 Conclusions 137
References 137
Chapter 7 Water Vapour Sorption of Natural Fibres 140

C. A. S. Hill

7.2 The Sorption Isotherm 141
x Contents
7.3 Water Sorption Models 144
7.4 Hysteresis 146
7.5 Water Adsorption and Cell Wall Swelling 148
7.6 Sorption and Heat of Wetting 149
7.7 Kinetics of Water Sorption 149
7.8 Water Sorption and Mechanical Behaviour 151
7.9 Methods to Reduce Water Sorption 155
7.10 Conclusions 157
Acknowledgement 158
References 158
Chapter 8 Environmentally Friendly Coupling Agents for Natural

Fibre Composites 161
R. Chollakup, W. Smitthipong and P. Suwanruji

8.2 Interface between the Natural Fibre and the Matrix 163
8.3 Types of Coupling Agents 166
8.3.1 Conventional Coupling Agents 167
8.3.2 Bio-based Coupling Agents 171
8.4 Pros and Cons of Using Bio-based Coupling Agents 176
8.5 Conclusions 177
Acknowledgement 177
References 177
Chapter 9 Probing Interfacial Interactions in Natural Fibre

Reinforced Biocomposites Using Colloidal Force
Microscopy 183
G. Raj, E. Balnois, C. Baley and Y. Grohens

9.2 Materials and Techniques 186
9.2.1 Preparation of Cellulose Thin Films 186
9.2.2 Preparation of Xyloglucan Thin Films 186
9.2.3 Preparation of Pectin Thin Films 187
9.2.4 Preparation of PLA Colloidal Probes 188
9.2.5 Atomic Force Microscopy 189
9.2.6 Microbonding Test 189
9.3 Results and Discussion 190
9.3.1 Model Surfaces 190
9.3.2 Colloidal Force Microscopy 191
9.3.3 Direct Force Measurements 192
9.3.4 Interfacial Shear Stress and Work of
Adhesion 197
9.3.5 Verifying the Nardin–Schultz Relation for
Flax/PLA Biocomposites 199
Contents xi
9.4 Conclusion 200
Acknowledgement 201
References 201
Chapter 10 Zein: Structure, Production, Film Properties and

Applications 204
Narpinder Singh, Sandeep Singh, Amritpal Kaur and
Mandeep Singh Bakshi

10.2 Structural Characteristics 205
10.3 Isolation and Production 207
10.4 Film Forming 210
10.5 Film Properties 211
10.6 Modifications 212
10.7 Applications 213
Acknowledgements 214
References 214
Chapter 11 Silk Fibre Composites 219

Panya Wongpanit, Orathai Pornsunthorntawee and
Ratana Rujiravanit
11.1 Introduction to Silk Fibres and their

Protein Components 219
11.1.1 Silk Fibres and their Sources 220
11.1.2 Physical Structure and Chemical
Composition of Silk Fibres 220
11.1.3 Characteristics of Silk Fibres and their
Protein Components 221
11.2 Fabrication and Potential Applications of Silk-based
Materials 222
11.2.1 Raw Silk Threads and Spun Silks 222
11.2.2 Regenerated Silk-based Materials 223
11.3 Silk-based Composites: Fabrication and
their Potential Applications 226
11.3.1 Silk Fibres as Composite Reinforcement 229
11.3.2 Silk Fibres as a Composite Matrix 238
11.3.3 Regenerated Silk as a Composite
Reinforcement 239
11.3.4 Regenerated Silk as a Composite Matrix 241
11.3.5 Miscellaneous 246
References 250
xii Contents
Chapter 12 Hybrid Composite Structures from Collagenous Wastes and
Environmental Friendly Polymers: Preparation, Properties
and Applications 257
M. Ashokkumar, P. Thanikaivelan and B. Chandrasekaran

12.1.1 Eco-benign Polymers 260
12.1.2 Bio-based Materials versus Environment 260
12.1.3 What are Biocomposites? 261
12.1.4 Value Added Products from Skin and
Leather Wastes: Current Trends 262
12.2 Biomaterials from Chromium-free
Proteinaceous Wastes 263
12.2.1 Formation of Hybrid Films Using Collagen
and Natural Polymers 264
12.2.2 Characteristics of Collagen/Biopolymer
Hybrid Films 265
12.3 Flexible Composite Sheets from
Chromium-containing Proteinaceous Wastes 266
12.3.1 Flexible Composite Sheet Formation 267
12.3.2 Properties of the Flexible Composite Sheets 268
12.4 Conclusion 270
Acknowledgement 270
References 271
Chapter 13 Spider Silk: The Toughest Natural Polymer 275

Gangqin Xu, Guoyang William Toh, Ning Du and
Xiang Yang Liu
13.1 Introduction to Natural Spider Silk Fibres 275

13.2 General Structure of Silks 276
13.3 The in vivo Formation of Silks 278
13.4 Mechanical Properties 279
13.4.1 Tensile Properties of Natural Silks 280
13.4.2 Factors Influencing the Mechanical
Properties of Natural Silks 281
13.5 Structure–Property Relationship 283
13.5.1 Structure Related to Mechanical Properties 283
13.5.2 Models 284
13.5.3 Techniques to Study the Structure of Silk
Fibres 288
13.6 Supercontraction 289
13.7 Silk Protein-based Novel Biomaterials 294
13.8 Summary 296
References 297
Contents xiii
Chapter 14 Mussel Byssus Fibres: A Tough Biopolymer 305
F. G. Torres, O. P. Troncoso and C. E. Torres

14.2 Natural History 306
14.2.1 Biological Functions 306
14.2.2 Mussel Anatomy 306
14.2.3 Mussel Species 307
14.3 Morphology and Structure 308
14.3.1 Chemical Composition 308
14.3.2 Macro Structure of Byssal Threads 309
14.3.3 Collagens Present in Mussel Byssus 310
14.4 Characterization 311
14.4.1 Structural Characterization 311
14.4.3 Thermal Properties 315
14.4.4 IR Analysis 317
14.4.5 Adhesive Properties 319
14.5 Potential Applications and Biomimetics 321
14.5.1 Biomimetics 321
14.5.2 Surgical Adhesives 322
14.5.3 Dental Adhesives 322
14.5.4 Biosensors and Immunosensors 323
14.5.5 Cell Culture and Tissue Engineering 323
14.5.6 Biotechnological Applications 325
References 326
Subject Index 330
Volume 2: Nanocomposites
Chapter 1 Nanocellulose: Potential Reinforcement in Composites 1

Alain Dufresne
1.1 Introduction 1
1.2 Hierarchical Structure of Lignocellulosic Fibers and
Mechanical Potential 2
1.3 Microfibrillated Cellulose 8
1.3.1 Preparation 8
1.3.2 Morphology 11
1.3.3 Pretreatments 12
xiv Contents
1.3.4 Aqueous Suspensions of MFC 14
1.3.5 Mechanical Properties of MFC Films 16
1.4 Cellulose Nanocrystals 18
1.4.1 Preparation 18
1.4.2 Morphology 19
1.4.3 Aqueous Suspensions of Cellulose Nanocrystals 22
1.4.4 Mechanical Properties of Cellulose
Nanocrystal Films 23
1.5 Conclusions 25
References 25
Chapter 2 Chitosan-based Nanocomposites 33

Sreejarani Kesavan Pillai and Suprakas Sinha Ray
2.1 Introduction 33
2.2 Structure and Properties of Chitosan 34
2.2.1 Importance of Chitosan 35
2.2.2 Chitosan-based Nanocomposites 36
2.2.3 Importance of Chitosan-based Nanocomposites 36
2.2.4 Types of Chitosan-based Nanocomposites 37
2.3 Structure and Properties of Montmorillonite 38
2.4 Nanocomposites: Preparation, Structure, and
Characterization 41
2.5 Chitosan/MMT Nanocomposites 43
2.5.1 Preparation Methods and Characterization 43
2.5.2 Properties 49
2.6 Applications of Chitosan/MMT Nanocomposites 59
2.7 Current Research Status and Future Scope 61
Acknowledgments 62
References 62
Chapter 3 Characterization of Molecular Interactions in Amylose

Starch Bionanocomposites 69
Deeptangshu S. Chaudhary
3.1 Introduction 69
3.2 Physical Factors Dominate Interactions 70
3.2.1 Common Plasticizers 70
3.2.2 Sodium Montmorillonite Nanoclay
Additives 71
3.2.3 Effect of Plasticizer Molecular Size 72
3.2.4 Effect of Plasticizer Molecular Symmetry 74
3.3 Intrinsic Chemistry Factors Dominate Interactions 75
3.3.1 Crystallinity 75
3.3.2 Crystallization Behaviour (Mechanism and
Growth) via the Avrami Equation 78
Contents xv
3.4 Physical–Chemical Interplay Dominates Interactions 80
3.5 Role of Moisture Molecules 81
3.5.1 DSC Analysis 82
3.6 Synergistic Interaction Maps 84
3.6.1 Stage One (Unsaturated, Relatively Loose,
Soft Polymeric Network) 84
3.6.2 Stage Two (Competitively Formed, Tight
Polymeric Network) 86
3.6.3 Stage Three (Saturated, Phase Separation
Polymeric Network) 87
References 89
Chapter 4 Soy Protein Nanocomposites: Emerging Trends and

Applications 91
Dagang Liu and Huafeng Tian
4.1 Introduction 91
4.2 Soy Protein/Inorganic Hybrid
Nanocomposites 93
4.2.1 Soy Protein/Clay Nanocomposites 93
4.2.2 Soy Protein/Carbon Nanotube Composites 96
4.2.3 Biomimetic Hybrid Soy Protein
Nanocomposites 97
4.2.4 Other Soy Protein/Inorganic
Nanocomposites 100
4.3 Soy Protein/Organic Nanocomposites 100
4.3.1 Soy Protein/Polysaccharide Nanocomposites 100
4.3.2 Soy Protein/Polymeric Emulsion Particle
Nanocomposites 103
4.4 Applications of Soy Protein Nanocomposites 103
4.4.1 Wood Adhesives 103
4.4.2 Fibers 104
4.4.3 Packaging Films 106
4.4.4 Reinforced Biodegradable Bioplastics 107
4.5 Conclusion 108
References 110
Chapter 5 Nacre from Mollusk Shells: Inspiration for

High-performance Nanocomposites 113
Reza Rabiei, Sacheen Bekah and Francois Barthelat
5.1 Overview and Microstructure of Nacre 113

5.2 The Deformation of Nacre 115
5.2.1 Experimental Studies 115
5.2.2 Analytical Elastic Models 117
xvi Contents
5.3 The Fracture of Nacre: Experimental Approach 120
5.3.1 Experimental Procedures 120
5.3.2 Crack Resistance Curves 122
5.3.3 In situ Microscopy Observations 123
5.3.4 In situ AFM Fracture Experiments 125
5.4 Theoretical Crack Resistance Measurement 127
5.4.1 Crack Bridging 128
5.4.2 Process Zone 129
5.5 Modeling the Deformation and Failure Patterns of the
Microstructure 131
5.5.1 Brittle Fracture of Tablets 134
5.5.2 Analytical Modeling of Tablet Junction
Failure 134
5.6 Optimum Structure and Length Scale 136
5.7 Large Microstructures for Columnar and Sheet Nacres 138
5.7.1 Perfectly Periodic Microstructures 138
5.7.2 Microstructures with Statistics and Large Scale
Waviness 139
5.8 Artificial Nacres 141
5.9 Conclusion 145
References 146
Chapter 6 Nanocomposites from Furanic Derivatives 150

Guan Gong
6.1 Furan and Furan Derivatives 150

6.1.1 Synthesis, Structure and Properties of
Furan 150
6.1.2 Furan Derivatives 151
6.2 Furfuryl Alcohol, Poly(furfuryl alcohol) and their
Applications 153
6.2.1 Furfuryl Alcohol Monomer and
Resinification 153
6.2.2 Application of FA and/or PFA in Porous
Carbon Materials 154
6.2.3 Application of FA and/or PFA in Carbon
Membranes 159
6.2.4 Application of FA and/or PFA in Carbon
Nanotube (CNT) Modification and PFA/CNT
Nanocomposites 163
6.2.5 Modification of Natural Fibers with FA
and/or PFA and Natural Fiber Reinforced
PFA Composites 169
6.2.6 Nanoclay or Cellulose Nanowhisker
Reinforced PFA Nanocomposites 173
Contents xvii
6.2.7Application of FA and/or PFA in Wood
Modification 177
6.3 Conclusions 180
References 180
Chapter 7 Starch Nanocomposites 185

Dipa Ray and Sonakshi Maiti

7.2 Starch 186
7.2.1 Source 186
7.2.2 Structure 186
7.2.3 Advantages and Disadvantages of Starch 191
7.3 Starch Nanocomposites 191
7.3.1 Starch Matrix with Different Nanofillers 191
7.3.2 Starch Nanoparticles with Different Polymer
Matrices 220
7.4 Biodegradability of Starch Nanocomposites 229
7.5 Application Potential 230
7.6 Limitations 230
7.7 Future Prospects 230
References 231
Chapter 8 Processing and Industrial Applications of Natural Polymer

Nanocomposites 234
X. Z. Tang, S. Alavi, K. P. Sandeep and P. Kumar

8.2 Preparation of Natural Polymer
Nanocomposites 236
8.2.1 Polymer/Layered Silicate
Nanocomposites 237
8.2.2 Carbon Nanotube-based
Nanocomposites 246
8.3 Industrial Applications of Natural Polymer
Nanocomposites 247
8.3.1 Food Packaging Applications 248
8.3.2 Biomedical Applications 250
8.4 Conclusions 251
Acknowledgments 252
References 252
xviii Contents
Chapter 9 Protein-based Polymer Nanocomposites for Regenerative
Medicine 255
Bibin Mathew Cherian, Gabriel Molina de Olyveira,
Ligia Maria Manzina Costa, Alcides Lopes Leão and
Sivoney Ferreira de Souza

9.2 Isolation of Protein-based Polymeric Fibers 257
9.2.1 Collagen 257
9.2.2 Gelatin 257
9.2.3 Elastin 258
9.2.4 Fibrin 258
9.3 Structure and Properties 259
9.3.1 Collagen 259
9.3.2 Gelatin 261
9.3.3 Elastin 262
9.3.4 Fibrinogen and Fibrin 263
9.4 Surface Modification of Protein-based Polymeric
Fibers 263
9.4.1 Material Approach 265
9.4.2 Biology Approach 266
9.5 Nanocomposite Preparation and
Characterization 267
9.5.1 Collagen Nanocomposites 267
9.5.2 Gelatin Nanocomposites 271
9.5.3 Elastin Nanocomposites 273
9.5.4 Fibrin Nanocomposites 278
9.6 Principles of Bioregenerative Engineering 280
9.6.1 Cell and Tissue Regenerative Fundamentals 280
9.6.2 Sources of Cells for Tissue Engineering 281
9.6.3 Cell Behavior with Nanostructures 283
9.7 Conclusion 285
References 286
Subject Index 294

CHAPTER 1
Natural Polymers: An Overview

MAYA JACOB JOHN*a,b AND SABU THOMAS c
a
CSIR Materials Science and Manufacturing, Polymers and Composites
Competence Area, P.O. Box 1124, Port Elizabeth 6000, South Africa;
b
Department of Textile Science, Faculty of Science, Nelson Mandela
Metropolitan University, P.O. Box 1600, Port Elizabeth 6000, South Africa;
c
School of Chemical Sciences, Mahatma Gandhi University, Priyadarshini
Hills P.O., Kerala, India
*Email: [email protected]
1.1 Introduction
The scarcity of natural polymers during the world war years led to the devel-
opment of synthetic polymers like nylon, acrylic, neoprene, styrene–butadiene
rubber (SBR) and polyethylene. The increasing popularity of synthetic poly-
mers is partly due to the fact that there are unlimited and economic avenues for
modification of chemical structures to obtain a product with specific properties.
However, this rampant use of petroleum products has created a twin dilemma:
depletion of petroleum resources (Figure 1.1) and entrapment of plastics in the
food chain and environment.1 The exhaustive use of petroleum-based resources
has initiated efforts to develop biodegradable plastics. This is based on
renewable bio-based plant and agricultural products that can compete in the
markets currently dominated by petroleum-based products. Table 1.1 presents
a selected list of the common synthetic polymers.
Another issue is that the disposal of plastics in landfills creates a serious
aesthetic problem in large urbanized areas of the world. The chemical stability
of plastic prevents plastic waste from decomposing into the environment at a

1
2 Chapter 1
Figure 1.1 Chart of world oil reserves.
Table 1.1 List of selected synthetic polymers.

Synthetic polymer
Poly(ethylene terephthalate)
Polyethylene
Poly(vinyl chloride)
Polypropylene
Polystyrene
Poly(tetrafluoroethylene)
Polyurethane
Polyamide
Polyacrylamide
rate comparable to the rate of waste generation. In the long run, the incentive
to preserve the local environment is reduced and the costs of cleaning and
recovery of contaminated sites rise. Large streams can also transport excess
plastic waste to other areas, creating a mobile contamination problem. Plastic
waste comprises 60–80% of the marine debris litter accumulated in ocean
shores. The problem of marine waste is aggravated by the low reliability of
removal mechanisms aimed at reducing marine plastic residual concentration in
the oceans. The effects of plastic waste on marine life include the entanglement
and ingestion of harmful plastics by marine vertebrates and the bioaccumula-
tion of toxicants along the food chain.
Natural polymers are those which are present in, or created by, living
organisms. These include polymers from renewable resources that can be
polymerized to create bio-plastics. There are two main types of natural poly-
mers: those that come from living organisms (these include carbohydrates and
proteins) and those which need to be polymerized but come from renewable
Natural Polymers: An Overview 3
resources (e.g. lactic acid and triglycerides). Both types are used in the pro-
duction of bio-plastics.
Among the different types of natural polymers, the best known resources
capable of making biodegradable plastics are starch and cellulose. Cellulose
is the most abundant carbohydrate in the world (40% of all organic matter is
cellulose). It is the main constituent of plants, serving to maintain their struc-
ture, and is also present in bacteria, fungi, algae and even in animals. Cellulose
from trees and cotton plants is a substitute for petroleum feedstocks to make
cellulose plastics.
Starch is a condensation polymer made up of hundreds of glucose mono-
mers, which release water molecules as they chemically combine. Starch is a
member of the basic food group of carbohydrates and is found in cereal grains
and potatoes. It is also referred to as a polysaccharide, because it is a polymer
of the monosaccharide glucose. Starch molecules include two types of glucose
polymers, i.e. amylose and amylopectin, the latter being the major starch
component in most plants, making up about three-quarters of the total starch
in wheat flour. Amylose is a straight-chain polymer with an average of about
200 glucose units per molecule. Starch is one of the least expensive biode-
gradable materials available in the world market today. It is a versatile polymer
with immense potential for use in non-food industries. The annual world
production of starch is well over 70 billion pounds weight, with much of it being
used for non-food purposes, like making paper, cardboard, textile sizing and
adhesives.
Chitin, a polysaccharide similar to cellulose, is Earth’s second most abundant
polysaccharide. It is present in the cell walls of fungi and is the fundamental
substance in the exoskeletons of crustaceans, insects and spiders. The structure
of chitin is identical to that of cellulose, except for the replacement of the OH
group on the C-2 carbon of each of the glucose units with an –NHCOCH3
group. The principal source of chitin is shellfish waste. Commercial uses of
chitin waste include the making of edible plastic food wraps and the cleaning up
of industrial wastewater.
Chitin is the main source of production of chitosan, which is used in a
number of applications, such as a flocculating agent, a wound healing agent, a
sizing and strengthening agent for paper, and a delivery vehicle for pharma-
ceuticals and genes. Chitin deacetylation leads to the formation of chitosan.
The process involves the use of strong alkali solutions for the removal of
N-acetyl groups, both at room and elevated temperatures. The amount of
chitin obtained annually from harvested shellfish is estimated to be over 39 000
tonnes. At least 10 billion tonnes of chitin are produced in the biosphere each
year, chiefly in marine environments.2
Collagen is one of the most plentiful proteins present in the bodies of
mammals, including humans. In fact, it makes up about 25% of the total
amount of proteins in the body. It has found increasing applications in tissue
engineering and repair.3 The ability of collagen to polymerize into a three-
dimensional fibrous matrix makes it an appealing material for extensive
therapeutic applications, including medical implants.4
4 Chapter 1
Table 1.2 List of common natural polymers.
Natural polymer
Polysaccharides
Starch
Cellulose
Chitin
Proteins
Collagen/gelatin
Casein, albumin, fibrogen, silks
Polyesters
Poly(hydroxyalkanoates)
Other polymers
Lignin
Lipids
Shellac
Natural rubber
Some of the other important natural polymers that are under scrutiny by the
research community, but beyond the scope of this book, include lignin, shellac
and natural rubber. In the category of natural polymers which need to be
polymerized is the interesting development of biodegradable plastics from
edible and non-edible vegetable oils like soybean oil, peanut oil, walnut oil,
sesame oil, sunflower oil, tung oil and castor oil.
Table 1.2 presents a selected list of the common natural polymers.5
The production of 100% bio-based materials as substitutes for petroleum-
based products is not an economical solution. Some of the possible solutions
are blending biopolymers with synthetic polymers and reinforcing natural
fibres with synthetic polymers (termed bio-composites), which are a viable
alternative to glass fibre composites.
1.2 Natural Polymer Research

The aim of this book is to examine the research conducted worldwide on the
use of different types of natural polymers. The book looks at the different
processing techniques of natural polymers as well as applications in advanced
industrial sectors. The structure, mechanical and thermal characteristics of
selected natural polymers are highlighted.
1.2.1 Natural Fibres

The history of fibre-reinforced plastics began in 1908 with cellulose fibre in
phenolics, later extending to urea and melamine and reaching commodity
status with glass fibre-reinforced plastics. Natural fibres are subdivided based
on their origins, coming from plants, animals or minerals. All plant fibres are
composed of cellulose, while animal fibres consist of proteins (hair, silk and
wool). Plant fibres include bast (or stem or soft sclerenchyma) fibres, leaf or
hard fibres, seed, fruit, wood, cereal straw and other grass fibres. Knowledge of
the structure of natural fibres is crucial in understanding the structural para-
meters (number, size and shape of cells, chemical constituents) and fracture
mechanisms in fibres.6
Some of the important natural fibres used as reinforcement in composites are
listed in Table 1.3.
Over the last few years, a number of researchers have been involved in
investigating the exploitation of natural fibres as load-bearing constituents in
Table 1.3 List of important natural fibres.

Fibre source Species Origin
Abaca Musa textilis Leaf
Agave Agave americana Leaf
Alfa Stippa tenacissima Grass
Bagasse – Grass
Bamboo (41250 species) Grass
Banana Musa indica Leaf
Broom root Muhlenbergia macroura Root
Cantala Agave cantala Leaf
Caroa Neoglaziovia variegata Leaf
China jute Abutilon theophrasti Stem
Coir Cocos nucifera Fruit
Cotton Gossypium spp. Seed
Curaua Ananas erectifolius Leaf
Date palm Phoenix dactylifera Leaf
Flax Linum usitatissimum Stem
Hemp Cannabis sativa Stem
Henequen Agave fourcroydes Leaf
Isora Helicteres isora Stem
Istle Samuela carnerosana Leaf
Jute Corchorus capsularis Stem
Kapok Ceiba pentranda Fruit
Kenaf Hibiscus cannabinus Stem
Kudzu Pueraria thunbergiana Stem
Mauritius hemp Furcraea gigantea Leaf
Nettle Urtica dioica Stem
Oil palm Elaeis guineensis Fruit
Piassava Attalea funifera Leaf
Pineapple Ananas comosus Leaf
Phormium Phormium tenas Leaf
Roselle Hibiscus sabdariffa Stem
Ramie Boehmeria nivea Stem
Sansevieria Sansevieria Leaf
(bowstring hemp)
Sisal Agave sisalana Leaf
Sponge gourd Luffa cylindrica Fruit
Straw (cereal) – Stalk
Sun hemp Crorolaria juncea Stem
Cadillo/urena Urena lobata Stem
Wood (410 000 species) Stem
6 Chapter 1
composite materials. The use of such materials in composites has increased due
to their relative cheapness, their ability to recycle and the fact that they can
compete well in terms of strength per weight of material.
Volume 1 focuses on different sources and applications of natural fibres. One
chapter deals with novel renewable sources from which natural fibres can be
extracted. Another chapter looks at relating the structural anisotropy of nat-
ural fibres to mechanical properties. One of the challenges of using natural
fibres in aerospace applications is the airworthiness requirements. Currently,
natural fibres are being explored for use in secondary structures in aircraft for
which flame, smoke and toxicity (FST) requirements are very stringent. This
has led to a lot of developmental research being undertaken in this field. A
further chapter therefore explores the flammability properties of natural fibre
reinforced composites. A crucial problem associated with the use of natural
fibres in composites is their hydrophilic properties. This aspect is dealt with in a
chapter on probing the water sorption characteristics of natural fibres. The
chemical modification of natural fibres has been well documented in the lit-
erature, but ideally it would also be desirable that the chemicals used for
modification should also be from renewable resources as it would preserve the
biodegradable nature of natural fibres. A chapter therefore focuses on envir-
onmentally friendly coupling agents for natural fibre-reinforced composites.
Other chapters include examining the characterization techniques of the
interfacial properties of natural fibre-reinforced composites and the increasing
applications of natural fibre composites in the automotive sector.
1.2.2 Protein Fibres

The book also deals with the properties of selected protein fibres. Protein fibres
are formed by natural animal sources through condensation of a-amino acids
to form repeating polyamide units with various substituents on the a-carbon
atom. The sequence and type of amino acids making up the individual protein
chains contribute to the overall properties of the resultant fibre.7 In general,
protein fibres possess moderate strength, resiliency and elasticity. They have
excellent moisture absorbency and transport characteristics and do not build
up static charge. Some of the common protein fibres include wool, spider silk,
cashmere, etc. Among natural fibres, silk exhibits exceptional properties,
especially in toughness and biocompatibility properties. A chapter therefore
focuses on the studies and properties of silk fibre-reinforced composites. Other
chapters include studies on collagenous waste-based composites and exploring
the properties and applications of mussel byssus fibres. Important advance-
ments in the field of zein fibres are also discussed in another chapter.
Volume 2 deals with the properties and characterization of selected natural
polymer nanocomposites. Cellulose nanowhiskers (CNWs) have emerged as
one of the most interesting bio-based nano-reinforcements in the last decade.8,9
Cellulose nanowhiskers can be generated from various plant sources with
transverse dimensions as small as 3–30 nm, giving a high surface-to-volume ratio.
It has also been shown that since the nanowhiskers are rod-like, they can be
self-assembled into chiral nematic liquid crystalline structures, not only in
solution but also in the dry state. The volume begins by exploring nanocellulose
as a potential reinforcement in composites. Chitosan (a natural polymer) is a
good candidate for the development of conventional and novel drug delivery
systems. Chitosan has been found to be used as a support material for gene
delivery, cell culture, and tissue engineering. However, practical use of chitosan
has been mainly confined to the unmodified forms. For a breakthrough in
utilization, especially in the field of controlled drug delivery, graft copoly-
merization onto chitosan will be a key point, which will introduce desired
properties and enlarge the field of the potential applications of chitosan
by choosing various types of side chains. The properties and applications of
chitosan and soy protein-based nanocomposites are discussed in subsequent
chapters. Other chapters include studies on furanic-based nanocomposites, the
characterization of molecular interactions in amylose/starch nanocomposites,
and unique properties of nacre from mollusc shells.10,11 The last two chapters
touch upon the industrial and biomedical applications of natural polymer
nanocomposites.
References
1. M. J. John and S. Thomas, Carbohydr. Polym., 2008, 71, 343–364.
2. K. D. Sturm and K. J. Hesse, Ocean Challenge, 2000, 10, 20.
3. D. A. Wahl and J. T. Czernuszka, Eur. Cells Mater., 2006, 11, 43–56.
4. K. Madhavan, D. Belchenko, A. Motta and W. Tan, Acta Biomater., 2010,
6, 1413–1422.
5. E. S. Stevens, Green Plastics, Princeton University Press, Princeton, 2002.
6. K. G. Satyanarayana and F. Wypych, in Handbook of Engineering Bio-
polymers: Homopolymers, Blends and Composites, ed. S. Fakirov and
D. Bhattacharyya, Hanser, Munich, 2007, pp. 3–47.
7. http://www.textileschool.com/School/Fiber/NaturalProteinFibers.aspx
8. K. Oksman, A. P. Mathew, D. Bondeson and I. Kvien, Compos. Sci.
Technol., 2006, 66, 2776–2784.
9. A. P. Mathew, A. Chakraborty, K. Oksman and M. Sain, in Cellulose
Nanocomposites: Processing, Characterization and Properties, ed. K. Oksman
and M. Sain, ACS Symposium Series 938, Oxford University Press,
Oxford, 2006, pp. 114–131.
10. R. K. Pai, L. Zhang, D. Nykpanchuk, M. Cotlet and C. S. Korach, Adv.
Eng. Mater., 2011, 13, 415–422.
11. A. Gandini, Macromolecules, 2008, 41, 24.
CHAPTER 2
Biomimetics: Inspiration from

the Structural Organization of
Biological Systems
KALPANA S. KATTI,* CHUNJU GU AND
DINESH R. KATTI
Department of Civil Engineering, North Dakota State University, Fargo,

ND 58105, USA
2.1 Introduction: Hierarchy and Structural Order

Structural materials found in nature exhibit hierarchical structures that span a
structural order over length scales from molecular to macroscopic.1–5 Optimi-
zation of properties and structural redundancy as well as growth are all rami-
fications of the hierarchical structures that result from eons of evolution. The
following sections elucidate the hierarchical details of structure in bone, teeth,
seashells and spider silk and fabrication attempts at duplication of these structures.
2.2 Biological Materials Systems

2.2.1 Bone
Bone is a remarkable biological material, providing skeletal stability, support
and protection of vital organs. The structure of bone has been extensively

8
Biomimetics: Inspiration from the Structural Organization of Biological Systems 9
studied historically as well as recently, owing to its perfect adaption of
mechanical properties to metabolic functions. In addition, as a protein/mineral
nanocomposite, bone combines the optimal properties of both components:
stiffness and toughness. This rather unusual combination of material properties
is a combination of rigidity and resistance against fracture.6 The unique
mechanical properties of bone and the structure–property relationship have
attracted significant attention.
Research on the structure of bone dates back to the early 17th century, when
the compound microscope was invented. Clopton Havers is generally credited
with providing the first description of the porous nature of bone in 1691, but
due to the poor quality of the magnifying lenses, the initial descriptions dealt
primarily with the canal system and the ‘‘laminar’’ structure of bone without
the presence of osteonal bone. In the 18th and 19th century, some observations
were described and defined in detail, such as the Haversian system of lamellae,
and the orientation and disposition of lacunae and canaliculi.7 By utilizing
polarized light microscopy, Schmidt found that the crystallographic ‘‘c’’ axis of
mineral in bone is well aligned with the collagen fibrils.8 Further detailed stu-
dies came after the invention of scanning electron microscopy (SEM) and
transmission electron microscopy (TEM) in 1930, which enabled examination
of structures on the nanometer scale. With the help of these high-resolution
instruments and other techniques such as X-ray diffraction, polarized optical
microscopy, sonic velocity, as well as mechanical tests, the hierarchical struc-
ture of bone was discovered and depicted. Atomic force microscopy (AFM),
which appeared in 1980, made it possible to investigate the structure of bone in
the ambient environment on the nanometer scale.9–11 Although the overall
structure from nano to macro scales of bone has been extensively studied, more
detailed research is also needed.
Bone refers to a family of materials having in common a basic building block,
the mineralized collagen fibril; however, the structural organization of the
fibrils is different in different bone types. The family of bone also contains
dentin, cementum and mineralized tendon, which have various proportions of
bone components.12 Disregarding the different shapes, bone is generally
mechanically divided into two types, compact bone and cancellous bone,13
which will be introduced later.
Primarily, bone tissue is composed of collagen fibres, crystals of a calcium
phosphate complex also known as carbonated apatite [Ca10(PO4,CO3)6(OH)2]
(namely, mineral) and a cement containing mucopolysaccharides, among other
biopolymers. The chemical analysis of bone shows that there are three primary
components in bone: collagen, mineral and water. The collagen part accounts
for nearly 1/3 and the mineral part accounts for nearly 2/3 of the dry weight of
bone matrix.14 The water component is about 10–12 wt% of cortical bone and
20% of the bone matrix.15 The mineral crystals and grow in the triple helical
collagen fibres and replace some of the water while mineralization takes place.
The bone structure has been described in terms of up to six or seven hierarchical
levels of organization from nanoscale collagen and mineral to macroscale
femur bone (Figure 2.1).12,16,17 Recently, new studies indicate hierarchical
10 Chapter 2
(a) (b) (c)
(f) (e) (d)
Figure 2.1 Hierarchical organization of a human femur bone from macro- to

nanoscale. (a) Macroscale organ level: human femur bone. (b) Macroscale
tissue level: osteon. (c) Microscopic level: bone lamellae (adapted from
Giraudguille17). (d) Mesoscopic level: fibre bundle. (e) Nanoscale level:
mineralized fibril. (f) Molecular level: collagen molecule and mineral
particle. (Adapted from Fratzl.16)
structures in the collagen molecule in bone with three levels of hierarchy within
the molecule.18
2.2.1.1 Level 1: Collagen Fibrils and Minerals

2.2.1.1.1 Collagen. Type I collagen molecules, also called triple helices, are
supercoiled assemblies of three polypeptide chains, two identical a1-chains
and one a2-chain, each with over 1000 amino acid residues. Collagen type I
accounts for nearly 90% of its total organic content. The main part of a col-
lagen chain consists of Gly-X-Y repeats, in which X and Y can be any amino
acid but are frequently proline and hydroxyproline. A triple-helical molecule is
cylindrically shaped, with an average diameter of about 1.5 nm and length of
300 nm.19 Besides the main helical part, collagen triple helices also comprise
short nonhelical end sequences called telopeptides with both N- and C-terminal
ends. Telopeptides account for 2% of the molecule and are critical for fibril
formation in a self-assembly process.20 Figure 2.2 shows a schematic of the
general structure and triple helical motifs of the collagen molecules.
(a)
(b)
Figure 2.2 Collagen molecules and intermolecular cross-linking. (a) Triple-helical

structural motif of collagen molecules (adapted from Giraud-Guille19).
(b) Lysyl oxidase cross-linking. (Adapted from Kadler et al.20)
During the formation of a fibril the collagen spontaneously self-assembles

into cross-striated fibrils that occur in the extracellular matrix of connective
tissues. The fibrils are stabilized by covalent cross-linking which is initiated by
oxidative de-amination of specific lysine and hydroxylysine residues in collagen
by lysyl oxidase.20 The intermolecular cross-linking provides the fibrillar
matrices with various mechanical properties, such as tensile strength and
viscoelasticity. Both high-performance liquid chromatography (HPLC) and
Fourier transform infrared spectroscopy (FTIR) have been frequently adopted
to analyze the cross-links.21
2.2.1.1.2 Mineral. The mineral in bone primarily consists of poorly crys-

talline non-stoichiometric carbonated hydroxyapatite (dahllite), which has a
plate-shaped hexagonal crystal structure. Bone crystallites are probably the
smallest biogenic crystals. They are only 1.5–4 nm thick, 25 nm wide and
50 nm long on average,12 but very little is understood about the atomic struc-
ture. The c-axis of the unit cells of these crystallites in bone is usually aligned
parallel with the long axis of the adjacent collagen fibres.22 Studies using
AFM indicate that many of these mineral plates appear to be aligned, form-
ing larger aggregates (475–600 nm long 75–90 nm thick) that also retain
collagen periodicity along their exposed edges.11
2.2.1.2 Level 2: Mineralized Fibril

The triple-helical collagen molecules assemble into the fibril in a staggered
arrangement. Within the fibril there is a linear shift of B67 nm (D-period)
between neighboring molecules. The D-period is divided by overlaps and gaps,
where an overlap is around 27 nm and a gap is 40 nm (shown in Figure 2.2b).
Robinson and Watson22,23 pioneered a TEM study and reported the 68-nm
12 Chapter 2
banding pattern in collagen fibrils. The assembly of collagen molecules into
fibrils is an entropy-driven process, driven by the loss of solvent molecules from
the surface of protein molecules, resulting in assemblies with a circular cross-
section, which minimizes the surface area/volume ratio of the final assembly.20
Mineralized fibrils are the basic building blocks of bone. Minerals are inti-
mately associated with the collagen framework in which they form, resulting in
a highly complex but ordered mineral/organic composite material. Studies of
crystal growth show that crystals are first formed in the gap, and then they
continue to grow and penetrate into the overlap zone, thus pushing aside the
triple-helical collagen molecules and even breaking cross-linking and other
bonds.24 Because the density of crystals is higher in the gap region, a periodic
mineral density profile with around 67 nm spacing can be observed by electron
microscopy. The width of a mineralized fibril is about 100 nm.
It is also important to mention that the spaces between mineralized collagen
fibrils (extrafibrillar) are filled with non-collageneous macromolecules and
minerals. The extrafibrillar volume is at least 60% of the total, while the fibrils
are no more than 40%. More of the mineral appears to be extrafibrillar than
within the fibrils, and is cemented together by non-collagenous organic matter.25
2.2.1.3 Level 3: Fibrillar Arrays

The mineralized fibrils are self-organized into fibre bundles along their length
and the bundles may fuse with neighboring bundles. How the mineralized fibrils
are aligned is of great interest, which leads to a great difference in mechanical
properties. Two arrangements have been suggested.12 One is an arrangement of
mineralized collagen fibrils aligned both with respect to the crystal layers and
the fibril axes. The other arrangement of mineralized collagen fibrils is with
only the fibril axes aligned. Sonic velocity measurements in three orthogonal
directions of macroscopic specimens show significant differences,26 implying
that orthotropic order at the fibril level may well extend to millimeter distances.
2.2.1.4 Level 4: Fibrillar Array Patterns

The fibril bundles continue to be organized into layers or lamellae with a few
microns thickness, and they in turn are arranged in a variety of ways into
higher-order structures, depending on the bone type. Four of the most common
patterns have been presented. In a parallel array pattern, all the fibrils are
parallel to each other. In a woven array pattern, individual fibrils or fibril
bundles are randomly organized. A radical fibril array is characteristic of the
bulk of dentine, which also belongs to the bone family.12 Further, plywood-like
structures are common in nature, which is believed to have a structure–function
relationship.19 SEM and TEM studies showed that lamellar bone is made up of
alternating collagen-rich and collagen-poor layers, all having an interwoven
arrangement of fibres.27 The successive layers in a thin/thick lamellar unit
proceed by an angle of roughly 301 from one layer to the next.28
2.2.1.5 Level 5: Osteon
The osteon, also called the Harversian system, is the fundamental structural
unit of compact bone. Each osteon consists of concentric layers or lamellae that
surround a central canal, the Haversian canal. The Haversian canal, parallel
to the long axis of the bone, contains the bone’s nerve and blood supplies.
Between adjoining osteons there are angular intervals that are occupied by
interstitial lamellae. These lamellae are remnants of osteons, the greater parts of
which have been destroyed.14 The osteon takes part in the remodeling process,
whereby tunnels are eroded and then filled in again with cylinders of bone.
2.2.1.6 Level 6: Compact and Cancellous Bone

Compact bone is solid, with the only porosities existing for canaliculi, osteocyte
lacunae, blood channels and erosion cavities, while cancellous bone has por-
osity that is easily visible to the naked eye. Overall, bone serves a protective
function by the construction of two compact plates separated by intervening
cancellous (spongy or trabecular) bone. This construction is beneficial for the
maximum absorption of energy with the minimum trauma to the bone itself.
2.2.1.7 Mechanical Advantages of the Hierarchical Structure

As pointed out by Gao et al.,29 bone is much less sensitive to flaws because of its
hierarchical structure. The hierarchical design distributes stresses throughout
the levels of a structure, thereby minimizing dangerous stress concentrations
that could precipitate failure and fracture. The hierarchical feature of bone
controls the fracture properties, particularly the toughness.13 Because the col-
lagen fibres in neighboring lamellae are oriented at some angle to each other,
fracture surfaces show considerable roughness. The work of driving a crack
across the interfaces consisting of the plates, sheets and Haversian systems of
bone is much greater than it would be if the material were homogeneous.30
Recently, many efforts have been made to model the structure of bone and
its constituents through continuum and molecular modeling methods. The
mechanical properties of bone and the relationship of the properties to the
hierarchal structural organization have been extensively investigated.29,31–41
Continuum methods, using finite element modeling (FEM), are used to cal-
culate the fundamental mechanical properties of bone.42–45 Attempts at mul-
tiscale modeling techniques for collagen have been made.18,36,46–50 Recent
experimental studies using fluorescence resonance energy transfer has indi-
cated molecular interactions between apatite and collagen. Simulations using
steered molecular dynamics have indicated the nature of the interactions
(primarily non-bonded), as well as their role on mechanics of the collagen
molecule.50,51 In addition, it has been shown that the load-deformation
of tropocollagen interacting with HAP50 is significantly influenced by water.
Molecular and nanomechanics of bone have also been described in a number of
studies5,18,34,52–55 and different nanoscale deformation mechanisms have been
14 Chapter 2
proposed in the literature. It has also been shown recently that the collagen
molecule, when observed in its full length through simulations, exhibits a
new level of hierarchy of structure which has significant contributions to the
mechanics. The three levels are: level 1, the helicity of the individual polypep-
tide chain; level 2, the helical organization of the three peptide chains; and a
third new hierarchy, the helicity of the overall triple helix that can only be
observed in a full length collagen molecule.18
2.2.2 Teeth
Teeth are small, calcified, structures found in the jaws (or mouths) of many
vertebrates that have the primary function of breaking down food. Teeth are
anchored within alveolus bone sockets and are held in place by a thin cemen-
tum interlayer adjoining the periodontal ligament. The part of the tooth that
projects into the mouth is called the tooth crown, while the part that is set into
the jaw is called its root.56 The tooth interior generally consists of three layers.
The outer layer of enamel (96 wt% mineral), which is the hardest tissue in the
body, covers part or all of the crown of the tooth. The middle layer of the tooth
is made up of dentine (or dentin), with an enamel/dentine junction (EDJ)
several micrometers thick separating it from enamel. Dentine is less hard than
enamel and similar in composition to bone (70 wt% mineral, 20% organic,
10% water). The third and the innermost layer is pulp, containing blood vessels
and nerves. The hard, brittle enamel coat protects the soft, tough dentine and
pulp interior.57
Enamel is not only hard (resistant to permanent surface deformation), but
also very tough (resistant to crack propagation and brittle fracture).58 Unlike
bone, which is a collagen-based ceramic composite, enamel is a tissue miner-
alized with calcium phosphate, containing no collagen or cells, but rather
having long, thin strands of hydroxyapatite (HAP) that are woven into a fabric-
like ceramic. Enamel retains less than 0.5% protein and a very small amount of
water and holds the mineral fibres together.59 Additionally, in contrast to bone,
enamel is not remodeled during its lifetime since the ectodermally derived cells
that create enamel are lost once the tooth erupts into the oral cavity.60
2.2.2.1 Hierarchical Structure of Enamel

The enamel mineral is a form of non-stoichiometric carbonated calcium
hydroxyapatite [Ca10(PO4)6(OH)2]. The Ca/P ratio of enamel apatite is slightly
lower than that of hydroxyapatite.61 Enamel exhibits a hierarchical organiza-
tion that spans nanoscale to macroscale levels. Some literature regards the
nanospheres as the first level, virtually as the first step of biomineralization. At
the nanoscale, amelogenin molecules undergo self-assembly to form 15–20 nm
spheres.60 In mature teeth, apatite crystallites are the least complex and smallest
structural unit.58 The crystals resemble long bars with a hexagonal cross-section
at a final size of about 30–40 nm across.56 Here we list the five specific levels of
structure based on structural complexity of mature teeth.62 These orders of
scale are interdependent and coalesce with one another, thereby creating a
structural continuum in the tooth organization.
2.2.2.1.1 Level 1: HAP Crystallites, a Few Ångstroms Wide. Unlike the

minute, short crystallites of bone, the crystallites of dahllite in enamel are
needle-like, narrow and extremely long. Reports of crystallite length vary from
a fraction of a micron to 100 mm. Each crystallite is surrounded by nanospheres
favoring growth on their c-axis and proper spacing among the crystallites.60
In the early mineralized enamel crystals, ribbon-like crystals appear near the
ameloblast at the DEJ. As maturation occurs, the enamel crystals become more
densely packed and more highly oriented.63 Cross sections of fully mineralized
crystallites are usually hexagonal64 or rhomboidal.65 Many other growth
habits, including rectangular or irregular crystals (30–40 nm in width) which
contain ‘‘notches’’, have also been observed.64 Simmer proposed a three-stage
enamel crystal growth mechanism which involved a crystal precursor, octa-
calcium phosphate (OCP). The three-stage mechanism includes: (1) formation
of the incipient seed of the crystals; (2) two-dimensional growth of the seed
(a- and b-axes); (3) three-dimensional growth, which involves growth along the
c-axis and then one-unit-cell thickness of OCP hydrolyzes to a two-unit-cell
thickness of HAP, thereby leading to a contraction of the lattice in one
direction and growth in the c-axis produces a regular hexagonal prism. Further,
enamel incorporates some ions in its apatite lattice, such as HPO42–, CO32–,
Na1, F–, etc., different from ideal hydroxyapatite. Therefore, some ions fit in
the interstices and cause distortions in the close-packed lattice.65
2.2.2.1.2 Level 2: Prisms. Concerning the arrangement and orientation of

crystallites, two major classes of mammalian enamel are defined: prismless
(also named as aprismatic, nonprismatic or preprism in various publications)
and prismatic enamel.58 Prismless enamel refers to a discontinuous structure
separated by distinct boundary planes, and prismatic enamel has the cross-
sections of prisms that are demarked by sheaths that usually have curved
outlines.62 In primates, as in most mammals, prismatic enamel is the domi-
nant component of enamel patterns and the ameloblast cells weave the
hydroxyapatite crystallites into bundles called ‘‘rods’’ that decussate one
another.60 The diameter of a prism ranges from 2 to 10 mm.58 The sheath,
where protein and water accumulate, is approximately 100 nm thick.57 The
cross-sectional morphology of the prism sheath was determined by the shape
of the Tomes process on the tip of the ameloblast,62 and various morpholo-
gies were shown, such as circular prisms with complete sheaths, highly
derived prisms with key-hole cross-sections, etc., but it is still not known if
the differences in morphology of the prism cross-sections affect functions.
2.2.2.1.3 Level 3: Enamel Types. Units of enamel in which the prisms have
similar orientations are defined as enamel types. The orientations of prisms
are independent of their cross-section morphology. In order to study the
16 Chapter 2
orientation of prisms, the enamel/dentine junction (EDJ) is regarded as a
reference plane.62 The two prismatic enamel types that occur in primate
enamel are radial enamel and decussating enamel.
In radial enamel, all prisms are roughly parallel to one another as they rise
radially from the enamel/dentine junction and occlusally toward the enamel
surface.58 Radial enamel can be distinguished into different subtypes by dif-
ferences in orientation of the interprismatic matrix (IPM) crystallites relative
to the prisms, which can vary from almost parallel to intersections at angles of
approximately 901.62
In decussating enamel, prisms are arranged in regularly organized, alter-
nating layers or groups that rise from the enamel/dentine junction to the sur-
face at different orientations.58 Decussation is manifested optically in tooth
sections as so-called Hunter–Schreger bands (HSBs), due to variations in light
reflection from differently oriented prism bundles. Hunter–Schreger bands,
like radial enamel, are commonly observed in mammalian enamel but vary
in degree from species to species.57 Complete crossing of prisms at 901 to those
in adjacent groups (‘‘true decussation’’) does not occur in all species. More
often, the angle between prisms in adjacent bands is less than 901, and changes
as prisms pursue a slightly sinuous course from the enamel/dentine junction to
the outer tooth surface. In primates and most other mammals the layers are
usually several prisms thick, but in some rodents each layer is only one prism
thick. The most common type of decussating enamel is horizontal decussation,
in which HSBs are stacked on top of one another from crown to root, with long
axes of prisms in adjacent horizontal layers of prisms extending toward the
outer enamel surface at different angles. This is the type of decussating enamel
found in primate teeth.58 Tangential enamel and irregular decussation of prism
bundles were also recognized in some molars of rodents.62
2.2.2.1.4 Level 4: Enamel Patterns. Enamel patterns refer to the arrange-

ment of the enamel types within a tooth. Normally, mammalian teeth are
capped with enamel that is composed of two or more enamel types and have
a characteristic distribution through the enamel of the crown. The enamel
pattern was designated ‘‘schmeltzmuster’’ by Koenigswald in 1977.62 In
rodents, the enamel pattern regularly includes an inner layer of HSBs and
the outer layer of radial enamel. In molars of small-bodied primates the
enamel pattern usually consists of an inner layer of radial enamel and a
much thinner outer layer of prismless parallel crystal. In larger primates the
enamel pattern is usually composed of an inner layer of horizontal HSBs, a
middle radial layer and an outer prismless layer.58
2.2.2.1.5 Level 5: Dentition. Dentition is the most complex and largest-

scale hierarchical level that describes the variation of enamel patterns from
tooth to tooth. Animals whose teeth are all of the same type, such as most
non-mammalian vertebrates, are said to have homodont dentition, whereas
those whose teeth differ morphologically are said to have heterodont
dentition. At the dentition level, primates appear to have relatively little var-
iation in enamel microstructure in comparison with rodents.58 In human
teeth, enamel is the most dense (96% mineral) close to the outer surface and
less dense (84% mineral) near the enamel/dentine junction.66
Further, each level of complexity only provides limited information on one
aspect of the total structure. The thorough analysis should also take the study
of systematic interrelationships and biomechanical functions into account.
Although protein and water occur only a very small amount in enamel, they
are crucial to the development and toughness of bone. Amelogenin and
enamelin are the main proteins present in enamel. Initially, amelogenins make
up about 90% of the protein and are most important. They are hydrophobic
and probably control crystal size and orientation.56 As the mineral crystals
grow in size during maturation, amelogenins disappear and then suck out vast
amounts of protein (degraded with proteases) and water. Enamelin is the lar-
gest known enamel protein and its expression is highly restricted to developing
teeth. There are also some other proteins that have been observed, such as
tuftelin, which may play a role in the nucleation of enamel crystallites.67
Enamelins and tuftelins in the spaces between mineral crystals also serve as a
‘‘glue’’,68 and some other proteins such as ameloblastin and enamelysin are
believed to be essential for proper enamel mineral formation.69 The water in
enamel was found to be bound in two different ways. A small part is very
loosely bound, whereas the greater part is firmly bound to the mineral phase.70
2.2.2.1.6 Dentine. Dentine forms most of the volume of the tooth (as
shown in Figure 2.3). Dentine contains tubules that intersect the EDJ
Figure 2.3 Schematic representation of the basic structure of the human tooth.
(Adapted from Lawn et al.57)
18 Chapter 2
approximately perpendicularly, enabling transport of nutrients from the pulp
through the tooth interior.57 Dentine is rather like bone, with about 48% of
its volume composed of mineral and has an organic matrix based on type I
collagen fibres.
The microstructure of dentine was also suggested to be hierarchical. The
mineral crystallites are needle-like near the pulp and the shape continuously
progresses to be plate-like near the DEJ.71 The edge view of plate-like mineral
crystallites in dentine is as long as in enamel (up to 100 nm),72 and with
thickness in the range 2–3.5 nm. The collagen fibrils, approximately 30% by
volume, are roughly 50–100 nm in diameter; they are randomly oriented in a
plane perpendicular to the direction of dentine formation. The whole orien-
tation of collagen seems to determine that of the crystals which initiate within
and around them. Like bone structure, distinct bands from collagen fibrils in
longitudinal sections of dentine were also seen by TEM72,73 and AFM.74
At a higher level of organization, dentine is regarded as a composite, with the
intertubular dentine as the matrix and the tubule lumens with their associated
cuffs of peritubular dentine forming the cylindrical fibre reinforcement. Peri-
tubular dentine exhibits hardness close to that of enamel.56 At the greatest
length scale are the effective, or continuum, properties of dentine.75
2.2.2.1.7 Mechanical Advantages of the Hierarchical Structure. From a

micro-structural point of view, enamel and dentine are both very hetero-
geneous tissues. There are obvious mechanical advantages of their intricate
make-up. At the cost of a low stiffness and the loss of some compressive
‘‘strength’’, dental tissues have much greater toughness than plain ceramics
and survive much repeated loading.56 Cracks spreading between adjacent
prisms may be stopped if the rods change direction periodically (decussa-
tion).76 The complex fabric that dissipates forces traveling through teeth also
protects them from fracture.58
Many studies have been done on the hierarchical complexity of enamel and
dentine. However, there is still a lot that remains to be done if we are to
understand the adaptive significance of mammalian dental form and the rela-
tions between the hierarchical structure and function; a better understanding
of their structure could definitely contribute to improving the processing of
artificial composite materials.
2.2.3 Seashells
Seashells are natural mineralized nanocomposites with hierarchical structures.
There are a large variety of seashells found in nature and they possess many
different morphological types of shell structures,77,78 but most of them are built
of two CaCO3 polymorphs, viz. an outer prismatic layer of rhombohedral
calcite and an inner nacreous layer of orthorhombic aragonite, which are
sandwiched by an organic matrix containing glycine- and alanine-rich proteins
and polysaccharides, thus forming complicated multilayered microstructures.79
The outer calcite layer is hard and deployed by seashells to prevent penetration
from the outside, but it is prone to brittle fracture. The thick inner layer of some
mollusk shells is composed of nacre, also known as mother of pearl. Nacre
is well known for its superior mechanical properties, particularly fracture
toughness: a 3000-fold enhanced fracture resistance compared to a single
crystal of aragonite, which is its major constituent.80 Nacre consists of more
than 95 wt% of aragonitic calcium carbonate (CaCO3), which is a ceramic, and
less than 5 wt% of organic material, primarily composed of proteins and
polysaccharides. The literature shows that the main strengthening and tough-
ening mechanisms of nacre are because of its unique microarchitecture.80–83
2.2.3.1 Hierarchical Structure of Seashell

Like bone and enamel, as well as many other biological materials, seashells
exhibit a hierarchical structure with at least six levels.84 The nacre structure is
described by interlocked ‘‘brick/mortar’’ architecture, in which mineral is the
brick and organic materials are the mortar.78 Hierarchical levels starting from
the molecular level are discussed here.
2.2.3.1.1 Level 1: Nanograins and Intracrystalline Matrix in an Aragonite

Tablet. AFM imaging has been used to indicate the presence of a nano-
grain structure in the nacre platelets.84,85 The organic material network is
called an intracrystalline matrix. As seen in Figure 2.4, an aragonitic tablet
in nacre is composed of many individual nanograins. The mean size of the
nanograins as reported in literature varies from about 32 nm84,86 to 38 nm87
1 µm
Figure 2.4 AFM image of a single nacre platelet showing the nanograin structure.
20 Chapter 2
88
to about 45 nm with extensions. Synchrotron spectromicroscopy and X-ray
and TEM experiments indicate that at least part of the intracrystalline
matrix is crystallized and responds like a ‘‘single crystal’’88 and that the
thickness of the ‘‘intracrystalline’’ matrix is about 4 nm.87
2.2.3.1.2 Level 2: Polygonal Platelet and Intercrystalline Organic Matrix.

At the micrometer level, the polygonal tablets are made of aragonite crystals
(orthorhombic; CaCO3), with a thickness of approximately 0.2–0.5 mm and a
width of about 5 mm (Figure 2.5). The tablets are closely packed and orga-
nized into a layer. The intercrystalline organic matrix separates the platelets
and has a thickness of about 20 nm. All the platelets on the same layer are
twin-related whether they share a boundary or not.82 Studies on the biomi-
neralization of nacre reveal that individual aragonite tablets are nucleated on
the underlying matrix sheet and grow rapidly in a direction perpendicular to
the shell surface (the crystallographic c-axis). Growth parallel to the lamina
follows after the tablet has reached its maximum thickness. The growing
crystal tablets are hexagons.89
Additionally, based on this nanostructure, it has long been established that a
platelet diffracts as a single crystal.88 The platelet was described as a ‘‘pseudo’’
single crystal, i.e. an organomineral composite made up of aragonite nano-
grains all coherently oriented according to diffraction and embedded in a
crystallized organic phase.88 The identified elastic properties of the ‘‘inter-
crystalline’’ organic matrix were found to be twice as high as those of the
‘‘intracrystalline’’ one.87
1 µm
Figure 2.5 SEM image of hexagonal aragonite platelets with an intercrystalline

organic matrix in-between.
2.2.3.1.3 Level 3: Platelet Layers and Interlamellar Organic Matrix. The
platelet layers in nacre are parallel organized and sandwiched between inter-
lamellar organic materials. The thickness of a platelet layer and an organic
matrix layer are 500 40 nm and 26 5 nm, respectively.90 This structure
is the traditional ‘‘brick/mortar’’ structure, but the platelet layers are not
smoothly arranged; there are some fine structures between the layers, as
described below.
Additionally, interlocking of the platelets, which results from penetration of
the platelet layers and rotation of the layers as they are stacked, is observed in
nacre.83 The ‘‘platelet interlocks’’, observed on the fracture surface by SEM,
contribute an important toughening mechanism for nacre (see Figure 2.6) and
the formation of platelet interlocks results from small rotation between layers
of platelets. Between platelet layers, nanoscale mineral columns through the
organic matrix layers were also observed by TEM. They are randomly dis-
tributed on the surface of the aragonite platelets and are called mineral bridges.
Their average diameter is 46 8 nm with a height of 26 nm.90 Moreover,
nanoscale asperities are also present on the surface of the platelets.91 Observing
the cross-section of a platelet layer more carefully, it was seen that the lamellae
are parallel and roughly equally spaced (about 50–80 nm) from one another
and that the spaces between the layers are filled with an electron-lucent organic
material that shows no discernible regular structure.92
The interlamellar sheets between the platelet layers are composed of several
layers of organic material. Thin layers of b-chitin are sandwiched between two
thicker layers of silk fibroin-like proteins. The fibre axis of the b-chitin and silk
proteins are perpendicular to each other and aligned with the a- and b-axes
of the aragonite platelets, respectively. This well-defined spatial relationship
Figure 2.6 SEM image showing interlocks on the fracture surface of the platelets.83
22 Chapter 2
between the substrate and overgrowth phase suggests an epitaxial mechanism
of nucleation.4,92 Furthermore, solid-state NMR investigations of nacre reveal
the presence of an amorphous surface layer around the aragonite platelets,
which has a thickness of 3–5 nm and which contains hydrogen carbonate
groups.93 Molecular modeling of the organic–mineral interactions has indi-
cated the significant role of nonbonded interactions on the mechanical response
of the organic material.94,95
2.2.3.1.4 Level 4: Lamellar Array Pattern. In the abalone shell, the inter-
tablet boundaries form tessellated bands perpendicular to the lamellae
boundaries; in other situations, such as in the pearl oyster, the inter-tablet
boundaries are distributed randomly.84 In fact, the crossed lamellar structure
is the most widespread structure in mollusks, in which the lamellae are orga-
nized in more hierarchical levels.96,97 For example, the crossed-lamellar layer
of the Strombus decorus persicus seashell is composed of three sub-layers
with differently oriented basic building blocks.96 Each of the three crossed-
lamellar sub-layers consists of the four orders of lamellar hierarchy, and each
order of hierarchy is built of the lower-order blocks. The smallest building
block of the crossed-lamellar layer, the 4th-order one, has a polygonal shape
of approximate dimensions 100 200 100 nm.96
2.2.3.1.5 Level 5: Mineral Mesolayers and Growth Bands. At the next hier-
archical level, the structure is still ‘‘brick/mortar’’ like, in which the ‘‘brick’’
is the mineral mesolayer with a thickness of approximately 300 mm and the
‘‘mortar’’ is the organic material (so-called ‘‘growth bands’’) with a thickness
of about 20 mm.98 The periodic growth arrests create mesolayers that can
also play an important role in the mechanical performance of nacre, acting
as powerful crack deflectors.99 The thin organic layers separating the meso-
layers are thought to be formed upon shell growth in periods of less
calcification.84
2.2.3.1.6 Level 6: Calcite Layer and Aragonite Layer. The longitudinal

cross-section of nacre-containing shells, such as the abalone shell, displays
two layers with distinct microstructures: a prismatic calcite layer (P) and an
inner nacreous aragonite layer (N). Such an arrangement provides an opti-
mal protective function of the shell:82 the outer layer prevents penetration of
the shell, while the nacreous layer is capable of dissipating mechanical energy
through inelastic dissipation.84
Aragonite and calcite have very similar crystal structures; they also have the
same composition. Both of them consist of alternating layers of calcium ions
and carbonate ions perpendicular to the c-axis (in the ab plane). In aragonite,
growth is preferred along the c-axis; thus, under conditions of normal tem-
perature and pressure, aragonite forms as thin needles (acicular crystals) that
do not generally grow into large crystals. Calcite, on the other hand, tends to
form larger crystals, but they are very brittle and cleave easily along its {10.4}
planes.4
2.2.3.1.7 Mechanical Advantages of the Hierarchical Structure. The basic

order in the seashell build-up hierarchy may be of great importance to the
superior mechanical properties of seashells owing to an enhanced ability for
crack arrest at inter-lamellar boundaries.96 Based on the hierarchical structure
(Figure 2.7), toughening mechanisms at multiple length scales have been pro-
posed to explain the significant improvement in performance. The negligible
role of nano-asperities100 and mineral bridges,90,100–102 the significant role of
interlocking,103 the unfolding of protein molecules104,105 and mineral–protein
interaction,94,95,106 as well as the brick morphology (waviness),107 have been
shown to significantly increase the toughness and fracture strength of nacre.
Additionally, each composition of nacre has a great influence on its prop-
erties. It was believed that the organic matrix was the key to nacre’s fracture
resistance.108,109 Studies also show that loss of water in nacre did not influence
the microstructure but caused a substantial decrease in stiffness. The nature of
water in nacre itself has been investigated using FTIR.110 An enhancement in
toughness by three orders of magnitude and a two-fold increase in hardness
have been reported for CaCO3-built seashells compared with non-biogenic
CaCO3 crystals.96
(b) (d) (g)
(a) (c) (e)
(f)
Figure 2.7 Hierarchical organization in seashells, showing at least six structural

levels: (a) a bivalve shell; (b) the prismatic calcite layer (P) and nacre (N);
(c)–(g) mesoscale, microscale and nanoscale structures.84
24 Chapter 2
2.2.4 Spider Silk
Over the past 150 years, numerous studies have been conducted on spider silk
and also many popular articles have been written about this amazing material.
Spider silk can simultaneously exhibit a combination of tensile properties,
including high breaking strength, high initial stiffness and high toughness;111
especially the unusual combination of high strength and extensibility is a
characteristic unavailable to date in synthetic materials and yet is attained in
nature with a relatively simple protein processed from water.112
All spiders produce and use a variety of silks, but the orb-weaving araneid
spiders appear to make the most diverse use of them and produce the orb-
web.113 Spiders produce at least seven different silks, each synthesized and spun
by separate silk glands and spinnerets, and each with specific properties that
appear to be optimized to perform different functional roles.114 All of the silks
are composed completely of protein and each undergoes an irreversible tran-
sition from a soluble to an insoluble form during processing,115 but they have
different mechanical properties because of different amino acid compositions
and molecular organizations.113
Owing to its size and accessibility, major ampullate silk (produced by the
major ampullate gland) has been the focus of most studies. The major
ampullate silk, also called spider frame silk or dragline silk, constitutes the
frame and radii of the orb-web as well as the dragline upon which the spider
lowers itself. It is stronger per unit weight than high tensile steel, approaching
the stiffness or strength of the super-strong polymeric material Kevlar, and also
having an elasticity of up to 35%.113 The exceptional mechanical properties of
major ampullate silk have attracted much attention from scientists in various
disciplines to investigate the molecular and structural origins of its mechanical
properties.
2.2.4.1 Hierarchical Structure of Spider Silk

Nature is always instructively economical in its achievement of macroscopic
material diversity through rearrangement of molecular, nanoscopic and
microscopic building blocks. Like the previously described natural materials,
spider silk also possesses a hierarchical structure (Figure 2.8) and here we
present the five structural levels in detail.
2.2.4.1.1 Level 1: Primary Structure of the Proteins. The primary structure

of the proteins is the linear sequence of amino acids that form the molecule.
The proteins are composed almost entirely of repetitive elements. Four types
of shared amino acid motifs are observed in all of the proteins and they are
recognized as amino acid sequence motifs: (1) GPGXX: GPGGX/GPGQQ;
(2) Ala rich: poly-Ala/poly-Gly-Ala; (3) GGX; and (4) a ‘‘spacer’’ sequence
with amino acids that do not conform to the typical amino acid composition
of spider silks.116 All members of the spider silk gene family that have been
described at the protein sequence level share a limited set of amino acid motifs.117
Figure 2.8 Hierarchical organization in a spider silk assembly. (i) Spider silk proteins
consist of repeats of amino acid sequences as structural motifs that self-
assemble into higher-level structures such as b-sheet crystals and elastic b-
spirals. The b-sheet structures further assemble into soft micelles with the
hydrophilic ends at the perimeter. With increasing protein concentration,
micelles transform into metastable liquid crystalline structures. (ii) More
ordered fibrils emerge from spinning ducts and form constructed webs or
cocoons. (Adapted from Kluge et al.128)
For example, spider major ampullate silk is composed of two proteins,

major ampullate spidroin 1 (MaSp1) and major ampullate spidroin 2 (MaSp2).
MaSp1 is further composed of poly-Ala/poly-Gly-Ala/GGX, where X typically
stands for alanine, tyrosine, leucine or glutamine, and MaSp2 contains GPGGY/
GPGQQ/poly-Ala.115,118
2.2.4.1.2 Level 2: Amino Acid Sequence Motifs and Secondary Structure.

Although it has been known for a long time that each of the seven glands
produced proteins with unique amino acid compositions, it was only in the
1970s that the amino acid sequence motifs in spider fibroins were eluci-
dated.119 Further, it is only in the past few years that an understanding has
emerged of the reasons for the unique mechanical properties that spider silk
possesses. In particular, the proteins that comprise the silks and their sequen-
ces have provided key information that relates directly to these properties.114
Each amino acid motif will consistently construct specific secondary and
tertiary structures in the fibre. The amino acids motifs are suggested to be
structural modules, with large internal repeats flanked by shorter (B100
amino acid) terminal domains (N- and C-termini).112
Secondary structure describes the local spatial arrangement of main-chain
atoms of any chosen segment of a polypeptide chain. The common types in
proteins include a-helix, b-sheet and b-turn. Hayashi et al.117 suggested that the
26 Chapter 2
different structures of the b-sheet regions could be the crystalline areas in the
major ampullate silk fibre. With interlocks between adjacent chains, poly-Ala
b-sheets have a higher binding energy than the poly-Gly-Ala regions, so that
the tensile strength of major ampullate silk (poly-Ala) is greater than that of
minor ampullate silk. Also, b-turn spirals made from periodically spaced
proline residues impart extensibility to major ampullate and flagelliform silks.
With at least 43 continuously linked b-turns in its spring-like spirals, flagelli-
form silk possess 200% extensibility compared to 35% extensibility of
major ampullate silk, which has at most nine continuous b-turns.117 All in
all, sequence motifs such as polyalanine (polyA) and poly(alanine-glycine)
(polyAG) (b-sheet-forming), GGX (31-helix), GXG (stiffness) and GPGXX
(b-turn spiral) are key components in different silks whose relative positioning
and arrangement are intimately tied with the end material properties.112
Additionally, the C-terminal domain also has a role in the alignment of
secondary structural features and acts as a molecular switch that controls fibre
assembly.120 Therefore, new materials with tailored properties are expected to
be achieved by controlling the arrangement of these building block sequence
motifs as well as terminal domains.
2.2.4.1.3 Level 3: Tertiary Structure: Crystal and Amorphous. Tertiary

structure refers to the overall three-dimensional arrangement of all atoms in
a protein. Guan proposed three different structural models for spider major
ampullate silk (Figure 2.9).121 In the first molecular model, nanocrystallites
are dispersed in an amorphous matrix.122 In the second model, highly orien-
ted and weakly oriented b-sheets coexist.123 In the third model, crystal
b-sheet regions containing alanine (red lines) and glycine (blue lines) are
(a) (b) (c)
Figure 2.9 Different structural models proposed for spider major ampullate silk.
(a) Nanocrystallites are dispersed in an amorphous matrix (proposed by
Termonia). (b) Highly oriented and weakly oriented b-sheets coexist
(proposed by Jelinski). (c) b-Sheet regions, containing alanine (solid lines)
and glycine (dashed lines), interweaved with predominantly glycine-rich
31-helical parts (curls) (adapted from Guan121).
interweaved with the glycine-rich amorphous portion, which has pre-
dominantly 31-helical parts (blue curls).124
It was also suggested that the spider silk fibres have a three-phase model
which is composed of the well-oriented crystals and the isotropic amorphous
material as well as the weakly oriented and partially ordered third phase.125,126
As for the crystals, it is well established that the alternation of glycine with
either alanine or serine causes this sequence spontaneously to form b-pleated
sheet crystals through natural physical cross-linking. As the most heavily stu-
died secondary structure of silks, crystalline b-sheets contribute to the high
tensile strength of silk fibres. Wide-angle X-ray diffraction (WAXD) of major
ampullate silk in Nephila clavipes indicates that the mean (minimum) crystal
dimensions are approximately 2 5 7 nm, the crystallinity is in the range
10–15%, and the crystals are strongly aligned with the fibre axis. However,
Gosline et al. suggested that the total crystal content is higher than the 10–15%
noted above, and predicted a crystal content of 20–25% based on mechanical
tests.127 In the semi-amorphous state, one-third is oriented while the rest is
isotropic.125 The semi-amorphous regions of silk are commonly made up of
b-spirals and helical structures and thus provide silk with elasticity.128 The
amorphous chains, which interconnect the crystals, are estimated to be 16–20
amino acid residues long.127 It was proposed that the poorly oriented crystal-
lites may be important in effectively coupling the highly oriented crystalline
domains and the amorphous regions to produce the exceptional material.123
Intermolecular forces are also used by major ampullate silk to self-assemble
into nanocomposites composed of b-sheet nanocrystals imbedded in an
amorphous matrix. Keten et al. used mechanical shearing simulations and
illustrated that the nanoscale behavior of silk protein assemblies is controlled
by the distinctly different secondary structure content and hydrogen bonding
in the crystalline and semi-amorphous regions.118 The fundamental stability
criteria for the hydrogen-bonded interactions between water and the amide
groups in protein chains was shown recently by using quantum mechanics
simulations.129 Although the primary, secondary and tertiary structures of
spider silk have been known for decades, understanding the complete primary
sequence, conformational structure of the repetitive amino acid motifs and
their assembling mechanism has only just begun.130,131
2.2.4.1.4 Level 4: Liquid Crystalline Phase. The liquid crystalline phase is

an intermediate state during natural silk processing. Liquid crystalline spin-
ning of spider silk offers desirable properties and makes it possible to effi-
ciently spin a thread from molecules as large as silk proteins. Specifically, in
the spider’s gland and duct the molecules seem to form a nematic phase132 or
a cholesteric liquid crystalline phase133 as the concentration is increased by
evaporation of water. They form a substance that flows as a liquid but main-
tains some of the orientational order characteristics of a crystal, with the
long axes of neighboring molecules aligned.134 Observations reveal that the
‘‘rods’’ constituting the liquid crystalline phase of silk are not individual
28 Chapter 2
molecules or molecular segments, but instead arise at the supermolecular
level. The rods are aggregates of essentially globular molecules, held together
by non-covalent associations.135
Liquid crystalline spinning of spider silk stores protein dope molecules in a
highly concentrated liquid crystalline state and prealigns the large molecules in
the unspun dope, which may reduce the formation of defects, and then extends
these in the spinning duct to form a supremely tough thread with minimal
forces.134
2.2.4.1.5 Level 5: Fibril. After external drawdown of the liquid crystalline

phase ‘‘spinning dope’’, the silk fibril forms. At the fibril level, in a cross-
section the fibres have a circular profile, with a delicate triple-layered
‘‘coat–skin–core’’ structure in Nephila spider silk136 that was observed as a
skin–core duality structure.137 This structure was also observed by AFM
experiments.138 The thin outer layer consists of higher electron density while
the inner material has slightly lower electron density and numerous cavities
of very low electron density.136 Microfibrils enclosing fine channels might
impart energy-dispersive properties by deflecting the tips of cracks forcing
their way across the thread.134
2.2.4.1.6 Mechanical Advantages of the Hierarchical Structure. Spider

major ampullate silk is one of the best biological materials known with
respect to stiffness and strength (see Table 2.1). The web material is opti-
mized to allow it to absorb a large amount of energy without breaking. The
other major component of the orb-web, the viscid silk, is extremely exten-
sible, breaking at extensions of 200% or more, and has a very low stiff-
ness.113 The hierarchical structure of the silk, i.e. the primary structure, the
amino acid motif, the secondary structure, the tertiary structure and a skin–
core duality structure in order, as well as the special liquid crystalline
Table 2.1 Mechanical properties of spider silks and some other fibre
materials.
Material Strength (MPa) Elongation (%)
Major ampullate silk (dragline silk)110 4000 35
Minor ampullate silk110 1000 5
Flagelligorm silk (viscid silk) 1000 4200
Kevlar 4000 5
Rubber 1 600
E-glass 3445 –a
S2-glass 4890 –a
C fibre 6370 –a
SWNT 13 000–53 000 B23
Human hair 180 –a
Nylon 66 75 5
a
Low values.
spinning process, is crucial in order for the silk to retain these exceptional
mechanical properties.
Another unique feature of major ampullate silks is the supercontraction
when exposed to water, and the process is reversible and repeatable. Super-
contraction refers to the ability of the material contracting to a fraction of its
original length in order to make the necessary space for the incoming solvent,
and the result of the process is a swollen gel-like phase with a volume that can
be 10 times larger than the dry network.139 Unstrained silks can retract to about
one-half of their original length in water. In this way the wetting-induced
contraction of spider silk tightens the web whenever the humidity is very high
and is able to restore the shape and tension of a slack web after deformation by
precipitation, wind or prey.140 Supercontraction was suggested to be the result
of a rearrangement of the GPGXX motif within the silk proteins, where X
represents one of a small subset of amino acids.141 The mechanical properties
change dramatically when the fibre is wet, owing to the formation of a
hydrogen-bonded network in the amorphous phase and hydrophobic effects, as
well as the fact that the pre-stress of the chains interconnecting the nanocrystals
is released.142,143
2.3 Biomimetic Design and Fabrication

Mimicking biomimetic model structures has been extensively studied in the
literature and many fabrication methods have been developed to duplicate the
intricate structures of these biological systems. The characterization routes to
the study of hierarchical structures as well as the duplication of these structures
to achieve unique properties has been of much interest.
Attempts to mimic the nacre structure, primarily the brick and mortar
architecture, are numerous. A self-assembly process that mimics the nano-
laminated structure is reported in the literature that results in a 100-fold
enhancement in properties using aragonite and biopolymers.144 The flat pearl-
like structure is also duplicated in silica and mica,145 as is the layer-by-layer
assembly.146,147 The nacre structure has also inspired the design of hybrid
materials based on natural polymers organized at the molecular level with
inorganic solids such as clay and silicates.148 Rapid assembly of inorganic
tablets of materials such as talc has also been attempted, again inspired by
nacre.149 Nacre-like films have been designed using nanoclay sheets with soft
polymer coatings with rapid self-assembly via simple processing routes.150
Electrophoretic deposition techniques have been shown to be able to success-
fully produce laminated high-quality structures and this technique has been
used to make nacre-like structures using polymers and nanoclays.151–153
Another strategy used is the template inhibition method that involves making
films from solution, using soluble macromolecules anchored on insoluble
matrices. This is called the template-inhibition strategy, and it consists of
mineral deposition from solution onto a well-ordered two-dimensional struc-
ture of a self-assembled film on solid or liquid substrates.154 This techniques
30 Chapter 2
155
was also used to develop thin film crystals of CaCO3 on chitosan. Most of
the current efforts on nacre mimicking have thus been targeted towards the
laminated structures and the alternating hard and soft components.
Silk is considered as a biomimetic model due to a potential large range of
mechanical properties, which are the result of both the semi-crystalline
nanostructure and also the degree of hydration.156 Silks are also being explored
as materials for bio-microfluidics.157 Spider silks have been investigated for a
variety of biomaterial applications, such as substitutes for high-performance
muscle,158 and also several biomedical applications, including drug delivery,
sutures and tissue engineering.159–162 Further, silk-mimetic proteins have been
fabricated using a variety of processing routes. These have included self-
assembly of a number of silk-protein inspired copolymers, b-sheet-rich fibres
or films from triblock copolymers, multiblock copolymers composed of
poly(isoprene) and Ala5-spacer-Ala5, and brush copolymers from acrylonitrile
and silk peptides.163,164 An extensive review on composite materials based on
silk proteins is reported by Hardy and Scheibel.160
Other important processing routes being investigated include the potential of
genetically engineered polypeptides with tailored selectivity as means of self-
assembly.165–167 Often these biomimetics efforts, based on genetic engineering,
result in potential for applications in distinctly different avenues, such as a
recent innovation in textile materials based on engineered silk proteins.168
These methodologies have a high potential to impact a large range of appli-
cations, from electronics to biomedicine.
Bone and teeth structures, although an inspiration, are not subjects of recent
studies on replacement materials for these tissues; most studies for replacement
of these hard tissues169 are based on replacement of function rather than
structure. Advances in tissue engineering170 are likely to facilitate further
progress, owing to advances in the knowledge of biomineralization routes as a
result of biomimetic efforts.
2.4 Perspectives on Future of Biomimetic Technologies

Innovative techniques ranging from electrospinning, genetic pathways, spin
coating, layer-by-layer deposition, dip coating, solvent evaporation, self-
assembly and others are utilized to mimic many aspects of the organization of
biological systems. Often, these attempts achieve a modest improvement in
properties and are used for the design of composites for a variety of applica-
tions, ranging from photonic to biomedical. Other studies have evaluated the
key roles of structural nuances of biological materials, such as the interlocks in
nacre, on the properties. Hence an interest in the design of the next generation
of composites that truly mimic biosystems would be of use for combination
with modeling routes to identify key elements of the structure in biosystems
that are particularly relevant to enhancement in properties. Current studies on
modeling the properties of these hierarchical structures have also benefitted
from advancements of computational resources. In addition, unique and
advanced characterization tools such as high-resolution electron microscopy,
photoacoustic spectroscopy and atomic force microscopy reveal unprecedented
details of structure. A combination of modeling and characterization abilities
are expected to strengthen efforts in biomimicking in the future.
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CHAPTER 3
Natural Fibres as Composite

Reinforcement Materials:
Description and New Sources
KARINE CHARLET
Clermont Université, IFMA, Institut Pascal, BP10448, 63000 Clermont

Ferrand, France;
CNRS, UMR 6602, Institut Pascal, 63177 Aubière Cedex, France
3.1 Introduction
Traditionally used in textile industries, natural vegetable fibres are increasingly
used or pinpointed as having potential in effective reinforcement materials of
polymer matrices. They exhibit several advantages, which explains the growing
interest of composite manufacturers (mainly in automobile and building
industries), wishing to replace the more commonly used glass fibres.1–3 A recent
survey revealed that about 90% of the fibres used as composite reinforcement
in Europe were glass fibres and, at most, only 3% were natural fibres.4 Another
research study claims that almost 40 million tons of natural cellulose fibres
could be obtained from already available agricultural by-products, while the
current total fibre consumption in the world is about 70 million tonnes.5
Moreover, it has been estimated that substituting synthetic fibres by natural
fibres in automotive composite parts would reduce the material weight by 30%
and their cost by 20%.6 According to the Canadian manager of the national
program on bio-products, the replacement of only 3.5 kg of glass fibres by
natural fibres in each car produced in North America would reduce greenhouse

37
38 Chapter 3
7
gas emissions by at least 500 million tonnes per year. This strategy is
encouraged by many governments worldwide through new legislation that
deals with the end-of-life handling of materials. For example, in Europe, the
deposition fraction of a vehicle is to be limited to 5% of its weight by 2015.8 In
2006, already 19 European countries had managed to reuse or recycle 80% of a
car.9 This promotes the use of natural and degradable fibres in composite
materials instead of synthetic and inert ones. This substitution is likely to be
feasible at a reasonable cost provided the processes and equipment used to
produce fibre reinforced composites would be the same. However, these
environmentally friendly materials also present some drawbacks that still limit
their use.
In this chapter, we will only focus on vegetable fibres (except wood), as
protein fibres will be discussed in another part. After a brief description of the
benefits and the difficulties brought about by the use of such fibres in composite
materials, a survey carried out on new sources of cellulosic fibres is presented.
This gathers information on natural fibres for which investigations as com-
posite reinforcements began only a few years ago and already reveal promising
characteristics (isora, vakka, artichoke, celery, switchgrass, etc.). This part is
presented as a comparison of these fibres with those of more ‘‘classical’’ fibres,
i.e. fibres that have been under study for a couple of years (flax, hemp, sisal,
jute, bamboo, ramie, cotton, etc.). The objective is to show that the field of
research of suitable, available and effective fibres expected to improve the
properties of polymers is far from being limited.
3.2 Advantages and Drawbacks of Natural Fibres as

Composite Reinforcement Materials
The first advantage of natural fibres is that they are available worldwide,
depending on the species concerned.10 In North America, the main produced
fibres come from the growing of cotton, hemp, flax, sorghum and switchgrass,
whereas in South America, sisal, abaca, curaua, pineapple, bagasse and ramie are
the most harvested vegetable fibres. Some African countries are well-known for
the production of kenaf, okra, sisal or rhectophyllum fibres. In Europe, hemp
and flax are the most developed forms of non-edible plants, but also sorghum
and china reed. Asia, thanks to the large variety of climates it offers, gathers
several types of vegetable fibre harvests: bamboo, abaca, sisal, kenaf, ramie, flax,
hemp, coir, jute, cotton, isora, vakka, okra and china reed are available more or
less locally in this part of the world. The worldwide extent of vegetable fibre
growing enables their cost to be relatively low (Table 3.1).
In parallel to these fibres, by-products resulting from traditional harvests or
even weeds have been pinpointed as possible local sources of cellulose fibres,
such as hop stems, nettle stems, cornstalks, artichoke stems, cotton stalks,
soybean straws, velvetleaf stems, etc. The inclusion in composites of fibres that
are currently considered as waste products brings the advantage of limiting the
required disposal areas and of giving value to products that were previously
Natural Fibres as Composite Reinforcement Materials 39
worth almost nothing. Table 3.1 summarizes the production data from various
survey reports on vegetable fibres.10–19
The added value that can result from the use and exploitation of such natural
fibres in polymer reinforcement is not only economic; it may also encourage the
development of local enterprises (and thus, of rural areas) that possess the
know-how in terms of extraction and treatment of these fibres. It may also
generate a (sometimes new) agricultural-based economy, especially when
dealing with by-product promotion.20
According to many parameters, such as the species or the position in the
plant, natural fibres generally exhibit large aspect ratios (defined as length/
diameter) that can vary from a few score, notably for by-products, to several
thousands for nettle, ramie or flax fibres (Table 3.2).15,21–45 This particularity
has an important influence on the mechanical properties of the derived com-
posites,46,47 as they provide a quasi-continuous reinforcement within the
composites: this enables a more homogeneous charge transfer between the
matrix and the fibres and hinders possible crack propagation. It is generally
considered that the minimum fibre aspect ratio needed to achieve optimum
composite properties must range between 100 and 200.48 These relatively long
fibres can also be cut to a defined length to ease the processing of composites,
such as injection.
Owing to the lightness of their constituents (mainly sugar polymers) and to their
microstructure, the density of natural fibres is relatively low (circa 1.4–1.5) com-
pared, for example, to that of E-glass fibres (Table 3.3).11,15,16,21–23,33,36,39–41,49–62
Table 3.1 Production and cost of natural fibres and of glass fibres (approx-
imate values for raw material, largely dependent on period, country
and stock size).
Fibre Production (Mt) Year Cost ($/kg)a Ref.
Abaca 0.07 2007 1.5–2.5 11,12
Coir 1.0 2005 0.2–0.4 13
Cotton 21.2 2003 1.5–2.2 10
Curaua 0.00015 2003 n.a. 14
E-glass 2.2 2003 1.2–1.8 10,11,15
Flax 0.75 2004 0.5–1.5 10,11
Flax 1.0 2007 n.a. 12
Hemp 0.08 2004 0.6–1.8 10,11
Hop 0.1 2007 n.a. 12
Jute 2.77 2004 0.35 10,11,16
Pineapple (Brazil) 0.02 2000s n.a. 15
Ramie 0.27 2004 1.5–2.4 10
Rice straw 10 2006 n.a. 17
Sisal 0.31 2003 0.6 10,11
Sisal 0.23 2007 n.a. 13
Switchgrass n.a. 1999 0.06–0.08 18
Vegetable fibresb 5.0 2003 0.2–1 10,15
Wheat straw 0.6 2004 n.a. 19
a
n.a. ¼ not available.
b
Except cotton, wood and straws.
40 Chapter 3
Table 3.2 Dimensions (mean value and/or [range]) and aspect ratio of natural
fibres.
Diameter, d Length, L Aspect ratio
Fibre (mm) (mm) (L/d) Ref.
Abaca 20 [6–46] [2–12] 350 21,22
Artichoke – [100–160] – 23
Bamboo 14 [7–40] 2.7 [1.5–4.4] 190 22,24
Banana [11–81] [0.9–5.5] 75 15,21,22
Bark of cotton stalk [10–20] [1–2] 50–200 25–27
Celery 260 – – 28
Coir [10–24] [0.3–1.2] 45 15,21,22,29,30
Cornstalk 27 0.8 30 31
Cotton 20 [10–45] 20 [2–64] 1000 15,21,22,29
Curaua 60 [20–130] – – 32,33
Flax 19 [5–76] 33 [4–140] 1700 21,22,24,34
Hemp 25 [3–51] 25 [8.3–55] 1000 10,24
Hop stems 16.5 2 120 35
Isora 10.1 1 100 36
Jute 20 [5–30] 2 [1–6] 100 15,22,24
Kapok 19 [10–35] 19 [7–35] 1000 22,24
Kenaf 21 [12–50] 5 [1.5–11] 240 21,22,24
Miscanthus 13.7 0.552 40 37
Nettle 20 [10–126] 50 [2–87] 2500 22,38,39
Pineapple leaf 20 [7–80] 8 [3–10] 400 22,24,40
Ramie 50 [5–126] 120 [40–260] 2400 15,21,22,24
Rhectophyllum 174 [100–250] 6 [5–8] 35 41
Rice straw 350 1.53 6 17
Sisal 20 [4–50] 3 [0.5–8] 150 15,21,22,24
Sorghum [8–15] 1.4 [0.4–3.4] 130 24
Soybean straw 15.6 1.5 100 42
Switchgrass 20 [10–30] 0.24 120 43,44
Velvet leaf 11.4 0.94 82 45
Actually, cellulose, their main constituent, has a specific weight around 1.56 g cm–3,
which varies according to the relative humidity.63–65 This slight drop in density
between the main constituent and the fibre can be explained by the presence of
lighter impurities (such as waxes and oils64), but especially by their porosity mainly
due to the lumen, an inner cavity that contains the cytoplasm during the fibre life
but which becomes empty when dead. This is why we can find in literature either
‘‘bulk’’ or ‘‘apparent’’ density, which is a global measurement of the fibre specific
weight, and ‘‘real’’ or ‘‘absolute’’ density, which only takes into account the weight
of the cell walls. The presence of the lumen is also responsible for the excellent
properties of natural fibres as thermal and acoustic insulators.15,66–69 This lightness
brings about advantages not only during the fabrication stage of the derived
composites, especially during transportation, but also during their life cycle; their
use, for example as an automotive part, may decrease the weight of the structure
and thus reduce fuel consumption.
The low density and particular microstructure of natural fibres are
known to be mainly responsible for their relatively good specific
Table 3.3 Specific weights of natural fibres and of glass fibres (apparent
densities in brackets).
Fibre Specific weight, r (g cm–3) Ref.
Abaca 1.4–1.5 (1.1–1.2) 11,21,22,49
Artichoke 1.58 (1.21) 23
Bamboo 0.6–1.5 22,50
Banana 1.3–1.35 (0.7) 21,22,50
Celery 1.5 (0.45–0.80) 51
Coir 1.15–1.5 11,22,52,53
Cotton 1.5–1.6 11,22,52–54
Curaua 1.4 (0.92–1.1) 22,32,33,55,56
Date 0.96–0.99 50
E-glass 2.5–2.6 11,15,16,52,53,57
Elephant grass 0.82–1.08 58
Flax 1.4–1.54 (1.38) 11,16,21,22,52–54,57
Hemp 1.4–1.6 11,16,22,52,54,57
Isora 1.35 36
Jute 1.3–1.52 (1.23) 11,16,21,22,52–54,57
Kenaf 1.2–1.4 22,57
Luffa (0.92) 59
Miscanthus 1.4 60
Nettle (0.72) 39
Pineapple leaf 1.44 40
Ramie 1.50–1.56 (1.44) 11,21,22,52,54
Rhectophyllum 0.95 41
Sisal 1.33–1.5 (1.2) 11,21,22,50,52,53,61
Switchgrass 0.74 62
Vakka 0.81 50
mechanical properties, sometimes higher than those of glass fibres (Table

3.4).11,15,16,21–23,25,28,31,32,35–42,45,50–58,70–72 Globally, the specific tensile strength
of plant fibres is between 1600 and 2950 MPa cm3 g–1, while their specific tensile
modulus varies from 10 to 130 GPa cm3 g–1.15 Actually, the elastic modulus of
cellulose is about 140 GPa,73 while that of hemicelluloses has been estimated at
8 GPa;74 both varying according to the relative humidity. It is also well known
that lignin increases the fibre structure stiffness by acting as a compatibilizer
between cellulose and hemicelluloses.66 Note that, in Table 3.4, some mechanical
data have been obtained by testing elementary fibres and others by testing
bundles of fibres. This is why the specific strength and Young’s modulus are
expressed either in MPa cm3 g–1 (for ultimate fibres) or in g denier–1 (for bundles).
The varying contents of the constituents in the fibres, gathered in Table
3.5,15,22,23,25,29,31,32,35,36,39,41–43,45,59–61,72,75–83 are one of the explanations for
the different and scattered mechanical properties among and within the dif-
ferent species. Actually, it is understandable, amongst other points, that the
higher the cellulosic content, the stiffer the fibre. Moreover, the particular
arrangement of the cellulose microfibrils, embedded under a helicoidal shape in
the amorphous matrix made of hemicelluloses, lignin and pectins, is partly
responsible for the deformation mode of natural fibres. Usually, the larger the
angle of the microfibrils with the fibre axis, called the microfibril angle
42 Chapter 3
Table 3.4 Specific mechanical properties of natural fibres or bundles and of
E-glass fibres.
Strength Failure Young’s modulus
Fibre (MPa cm3 g–1) strain (%) (GPa cm3 g–1) Ref.
Abaca 414 4 34 11,22
Artichoke 50–220 – 3–11 23
Bamboo 383–552 1.4–1.7 22–39 22,50
Banana 450 3.4–4 13–15 22,50
Celery 23–167 2–5.7 1–5 28,51
Coir 145–174 15–46 3–5 11,22,28,52,53
Cotton 265–290 3–10 5–8 11,22,52,53
Curaua 360–1000 3–4.3 8.4–36 22,32,55,56
Date (base) 478 24 2 50
Date (leaf) 312 2.7 11 50
E-glass 940–1350 2.5–3.4 28–30 11,15,16,52,53,57
Elephant grass 226–272 2.5–2.8 9–10 58
Flax 240–1070 1.2–3.3 26–76 11,16,22,52,53,57,70
Hemp 210–1264 1.6–3 24–50 11,16,22,52,53,57
Isora 370–440 5–6 13–15 36
Jute 270–650 1.2–2.0 7–39 11,16,22,52,53,57
Kapok 41 2 2 22
Kénaf 538 3 12–42 22,57
Miscanthus 652 – 42.5 37
Nettle 900–2200 1.7–2.1 53–121 22,38,57
Pineapple leaf 287–1130 1.6–2 24–57 22,40
Ramie 330–610 2–3.8 29–43 11,22,52,53
Rhectophyllum 588 27.5 6.1 41
Sisal 340–530 2–5.5 6–30 11,22,50,52,53
Switchgrass 60–88 – 6 71,72
Vakka 678 3.5 20 50
Strength Failure Young’s modulus
Fibre (g denier–1) strain (%) (g denier–1) Ref.
Bark of cotton stalk 2.9 3.0 144 25
Coir – 16 50 21
Cornstalk 2.2 2.2 127 31
Cotton 3.5 8.0 44 54
Flax 6.0 2–3 215–300 21,54
Hemp 6.3 1–6 225–285 22,54
Hop stems 4.1 3.3 160 35
Jute 4.2 1.2–1.5 200–350 22,54
Nettle 0.22–0.57 2.3–2.6 – 39
Ramie 7.0 4–4.4 160–170 22,54
Sisal – 2–3 290 22
Soybean straw 2.7 3.9 94 42
Velvet leaf 2.9 2.5 194 45
(Table 3.619,22,25,31,35,36,38,41,42,45,57,59,79,84–95), the higher the failure strain, since

the fibrils will be able to twist when stretched.96,97 Nevertheless, this is not a
universal rule since, in some fibres, microfibrils may lie in different directions in
the cell walls. For example, in cotton, the microfibril angle is largely dependent
Table 3.5 Biochemical composition of natural fibres, in % (w/w) on dry fibres.
Fibre Cellulose Hemicelluloses Lignin Ash Ref.
Abaca 56–70.2 15–21.7 5.6–10 3 22,29,75
Artichoke 75.3 – 4.3 2.2 23
Bamboo 26–43 15–26 21–31 1.7–5 22,29
Banana 60–65 15 5–10 1.2 15,22
Bark of cotton stalka 79 – 14 0.5 25
Coir 35–44 0.2–15.4 33–45 – 15,75
Cornstalka 81 – 8.4 – 31
Cotton 90–95 1–6.3 0.7–1.6 1.4 15,22,29,75,76
Curaua 73–73.6 10–20 1.5–7.5 1.0 22,32,77
Date (leaf) 33.5 28.5 26.5 6.5 78
Flax 62–72 17–18.5 2–5 1.5 15,22,75
Hemp 63–78 5–16 3–6 1.5 22,75,79
Hop stema 84 – 6 2.0 35
Isora 74.8 – 23.0 1.0 36
Jute 59–71 13.3–15 10–13 0.4 15,22,75
Kenaf 37–57 18–26 15–21 2–5 22,29
Luffa 63 19.4 11.2 0.4 59
Miscanthus 43.1 26.7 22.1 3.9 60
Nettle 79–80 10–12.5 0.5–3.8 – 22,39
Okra 60–70 15–20 5–10 – 80
Pineapple 80–83 12.3–17 3.5–12 – 15,22,81
Ramie 72–85 14 0.5–0.7 0.3 15,22
Rice straw 44.3 33.5 20.4 – 82
Rhectophyllum 68.2 16.0 15.6 – 41
Sisal 60–78 12–13 8–14 0.3 15,22,61,75
Soybean strawa 85 – 11.8 1.0 42
Straw (rice, wheat) 30–50 25–30 15–20 5–10 29
Switchgrass 29–43 26–36 12–24 2–5 43,72,83
Velvet leafa 69 – 17 3.2 45
a
Samples treated with sodium hydroxide.
on the maturity of the plant (from 11 to 451),22,31,76,98 whereas bamboo fibres

possess several concentric layers (more than other bast fibres), i.e. several
microfibril angles.99,100
Another advantage linked to the lightness and the good mechanical prop-
erties of natural fibres is that their use as composite reinforcement requires less
polymer matrices. Actually, to obtain the same properties with natural fibre
reinforced plastics as with glass fibre reinforced plastics, a simple law-of-mix-
ture says that the volume fraction of natural fibres has to be higher than that of
synthetic fibres. This reduces the required volume fraction of the polymer
matrix, which is a positive thing when dealing with a petrochemical-based
product.
The plants from which the fibres described in the following are taken have
the advantage of being renewable, generally quite rapidly. Except for some
species such as bamboo or china reed, which reach maturity only after several
years, vegetable fibres are often extracted from annual plants. This particu-
larity, compared for example to wood fibres, acts in favour of the very current
concern of sustainable development.
44 Chapter 3
Table 3.6 Microfibril angle and crystallinity of cellulose in natural fibres.
Fibre Microfibril angle (1) Crystallinity (%) Ref.
Abaca 22.5 52 84,85
Banana 10–12 45–55 19,22,85,86
Bark of cotton stalk 15 47 25
Coir 30–49 25–33 19,22,84,87
Cornstalk 10.9 52 31
Curaua – 76 22
Flax 5–11 70 22,84,88–90
Hemp 2–7.5 60–88 22,57,79,84,91
Hop stems 8 44 35
Isora 20–26 – 36
Jute 7–10 78 22,57,92
Luffa – 59 59
Nettle 3 – 38
Pineapple 6–18 44–75 19,22
Ramie 3–12 61–70 22,84,85,88,93
Rhectophyllum 40 – 41
Rice straw – 40 19
Sisal 10–25 55–71 22,57,94,95
Soybean straw 12 47 42
Switchgrass – 51 31
Velvet leaf 18 38 45
Wheat straw – 55–65 19
Furthermore, vegetable fibres are known to be CO2 neutral, since their

composting or combustion does not release into the atmosphere their excess in
carbon dioxide (captured through photosynthesis to produce the sugars of their
skeleton).66 Their production and their extraction generally require low
amounts of energy, according to the species considered, compared to the fab-
rication of synthetic fibres which consumes high levels of energy (mainly
through heating).
Last but not least, natural fibres are less abrasive to mixing compared with
traditional glass fibres.14,33,66 This leads to advantages when dealing with
processing or recycling of natural fibre-based composites.91
However, vegetable fibres exhibit several drawbacks that are more or less
handicaps for their further development as polymer reinforcements. First of all,
as can be seen in the tables which gather the fibre properties (Tables 3.2–3.7),
there is generally a broad dispersion of the values. This is true not only for
mechanical properties and dimensions, but also for the composition contents.
Some works have already underlined this aspect of the fibres and explained its
origin by the numerous variations undergone by the fibres during their growth
(humidity, sunlight, wind, etc.). Other sources of property scattering are the
level of maturity, the position in the plant and the extraction pro-
cesses.29,66,101,102 This results in a large variation in fibre quality according to
global parameters, even if the methods used to measure them are often the
same. For example, the determination of the cellulose, hemicelluloses and lignin
contents is almost always obtained using the Van Soest and Wine method
103
developed in 1967. Nevertheless, the cellulose contents often vary by several
percent for a given species (see Table 3.5). Concerning the mechanical prop-
erties, attempts have been made to calculate a property that could help com-
pare the fibres properly (such as a calculated E-modulus),104 even if this does
not really reduce the scattering. Both the fibre stiffness and strength are directly
linked with their growth environment, the former being more dependent on the
cellulose content and quality, and the latter on structural defects (generally
called kink bands). Since enzymatic ‘‘accidents’’ can occur during the forma-
tion of the long polymeric chains, the dispersion in the biochemical contents
and thus in the morphological parameters cannot be controlled (and so, lim-
ited) during cell life. As far as flaws generated by the extraction processes are
concerned, they can be reduced both in amount and in dangerousness, as
proved by several studies comparing the properties of fibres extracted by hand
with those of fibres extracted mechanically: a large improvement in strength
was highlighted, but no reduction in scattering.105
With the aim of improving the reliability of the properties of fibres, several
authors have used statistical analysis, mainly the Weibull equation, to refine
their description.80,106–110 Whereas this kind of numerical method can help in
understanding the effect of fibre length on its strength and modulus, it does not
limit the range of properties.
Unlike synthetic fibres that are standardized and whose properties are gen-
erally very accurate, natural fibres suffer from their lack of reliability. This is
particularly incapacitating when trying to model the fibre or the derived
composite behaviour. As composite manufacturers expect reproducible
mechanical characteristics, the only way to ensure an effective quality level is to
take large security coefficients on the fibre properties, which in turn reduces
their attractiveness and prevents full exploitation of their advantages.
Another important characteristic of vegetable fibres is that they
absorb water very easily compared with most synthetic fibres
(Table 3.7).19,21–23,25,29,31,32,35,42,45,50,57,60,72,80,83,111,112 This is likely to be due
not only to the strongly polarized hydroxyl groups present at the surface of the
natural fibres, but also to the non-crystalline (and less-oriented) regions present
in their cell walls. Actually, as seen in Table 3.6, these non crystalline parts can
reach 60% of the fibre volume. They mainly comprise lignin and hemicelluloses
but also some cellulose. However, only the latter two are decisive in relation to
moisture behaviour since they are highly hydrophilic whereas lignin is
hydrophobic.21
The first problem generated by this property is a noticeable dimensional
instability, especially in the transverse direction of the fibre. Values as high as
45% of volume swelling have been reported for jute and abaca, but they gen-
erally range between 30% and 40%.21,22,54 This can raise problems, not only
during the processing stage, but also during the life of the derived composite.
Actually, during fabrication, low humidity levels may cause a non-uniform
dispersion of the fibres within the matrix.66 Moreover, once the composite is
processed, a small change in relative humidity of the atmosphere can lead to
fibre/matrix decohesion, i.e. initiating the collapse of the structure. To reduce
46 Chapter 3
Table 3.7 Moisture regain for some natural fibres (21 1C, 65% RH).
Fibre Moisture regain (%) Ref.
Abaca 5–14 21,22,29
Artichoke 5 23
Bamboo 9–10 50
Banana 9–15 19,21,22,29,50
Bark of cotton stalk 8.8 25
Coir 10–13 19,21,29
Cornstalk 7.9 31
Cotton 7–25 21,22,29
Curaua 9–12 32,111
Date 9–11 50
Flax 7–12 21,22,29,57
Hemp 6–12 21,22,29,57
Hop stems 8.3 35
Isora 5–8 112
Jute 8.5–17 21,22,29,57
Kenaf 9.5–17 22,31,45
Miscanthus 11 60
Nettle 11–17 22
Okra 8 80
Pineapple 10–13 19,22,29
Ramie 7.5–17 22,29
Rice straw 6.5 19
Sisal 10–22 21,22,29,50
Soybean straw 11.2 42
Switchgrass 6–12 72,83
Vakka 12 50
Velvetleaf 9.8 45
Wheat straw 10 19
the sensitivity of the natural fibres to moisture, some surface treatments have
been successfully applied and enabled a relative stabilization of the reinforce-
ment (see Chapter 8). Nevertheless, moisture does not play in favour of
developing such eco-composites, especially in applications where the variation
in humidity is high.
In addition, natural fibres are highly temperature-dependent: their proper-
ties, especially the mechanical ones, drop dramatically when the temperature
increases. Previous studies reported a temperature threshold for natural fibre
use as low as 200 1C, since the fibres show damage above that.48,66,80,113–116
Generally, degradation is almost complete around 350 1C, as shown by ther-
mogravimetric analysis for okra, artichoke, curaua, kenaf, jute, isora, luffa,
sisal, bamboo and hemp fibres.17,32,59,80,112,117,118 This low thermal stability
becomes a problem when processing composites. Actually, natural fibre com-
posites are mainly processed by thermoforming, which requires relatively high
temperatures (according to the thermal and rheological properties of the
polymer) to ensure low viscosity and good impregnation and/or dispersion of
the reinforcement. This acts as an important ageing factor of both the polymer
and the fibres, and causes premature damage in the derived composite.
When dealing with long fibre composites, fibre entanglement within the
composite may occur, especially during processing. This is particularly true of
natural fibres, due to the porosity present within the bundles, i.e. between
fibres. Madsen and Lilholt119,120 showed that, according to the processing
conditions and the species considered, there was an optimal fibre weight con-
tent in the derived composite, since increasing this content also increases the
porosity and thus reduces the mechanical properties. Their studies proved that
porosity has to be taken into account, particularly when designing plant fibre-
based rather than synthetic fibre-based composites. Moreover, this particu-
larity may prevent plant fibre-based composites from reaching the same
mechanical properties as highly concentrated glass fibre reinforced composites,
which goes against eco-composites development.
At the scale of the composites, the interfaces between the natural fibres and
the polymer matrix are generally weak, and many treatments have been applied
to the fibres to enhance compatibility between these materials and the
mechanical properties of the derived composites. More details about that
aspect will be found in Chapter 8.
Focusing on vegetable fibres, they are often naturally occurring in the shape
of bundles of fibres. This implies that another natural material enters the fibre
composition in the form of sandwiching cement (Figure 3.1). In the example of
flax, several studies underlined the fact that the middle lamella was a weak link
within a bundle,121 and it has recently been shown that the interfacial strength
between two fibres was at least twice as low as the interfacial strength between
elementary fibres and classical polymer matrices.122 As a consequence, this
matrix
bundle
fibre
fibre/fibre
interface
fibre/matrix
interface
Figure 3.1 Schematic representation of the interfaces within a natural fibre-based

composite.
48 Chapter 3
cement, which is generally made of pectins, lignin and waxes in contents that
differ according to the species considered,123,124 contributes significantly to
mechanical weakness. An understanding of the mechanical behaviour of this
phase would help to enhance the reliability of derived composites.
Added to the other drawbacks of natural fibres, their rot sensitivity and their
not-so-pleasant odour have also already been underlined.66,101 Concerning this
last point, Müssig et al. highlighted the role of pectins in this problem, based on
previous studies.22,125 They showed that the elimination of pectins from hemp
bundles increased the odour intensity caused mainly by mildew contamination
or overheating, since the fibre surfaces available for these gaseous reactions
were more numerous. They also proved that enzymatic treatment of bundles
led to a reduction of the odour intensity, especially at low temperatures, and
that, after a few months, this intensity returned to its initial level. Although
synthetic fibres do not exhibit this kind of problem, more investigations must be
carried out to convince potential users that odour should not get in the way of
natural fibre composites development.
A more economic problem that could be posed when dealing with plant-
based composites is their long term supply. The recent controversies around the
use of fields to produce plants for other applications (mainly for bioethanol),
rather than for feeding people, may raise the question of investing in new
technologies to produce eco-composites, since geopolitical aspects are likely to
intervene in the debate. This may also curb the industrial enthusiasm in shifting
to more environmentally friendly materials, even if this shift is largely
encouraged by several governments.
3.3 Description of New Sources of Vegetable Fibres

The fibres whose properties are described in this chapter come from different
sources. They are mainly extracted either from leaves, from stems or from
seeds, and some plants even possess several types of fibres, such as cotton: the
most exploited cotton fibres come from their seeds, but others can be found in
the bark of their stems. The most common cellulosic fibres (flax, hemp, ramie,
cotton, jute, kenaf) used to strengthen polymers have already been described
elsewhere;21,29,57,66 thus we will focus here on fibres that gave rise to recent
interest as composite reinforcement materials (Figure 3.2).
Among the new sources of fibres, isora fibres, which fill in the bark of
Helicteres isora plants, have gained a relatively great importance due to their
good mechanical properties. This plant is mainly harvested in East and South
India. The fibres are extracted from the plant by retting and look like jute,36 but
fibre production is still at a local level. Elementary isora fibres are, as in many
other vegetable plants, gathered in bundles that appear relatively porous, which
is an advantage when combined in a polymer matrix (Figure 3.3a).36,112 When
added to natural rubber in a 1:1 proportion, isora fibres enhance the stiffness of
the matrix by almost 30%.36 Several chemical treatments applied to these fibres
(acetylation, benzoylation, silane treatment) changed the morphology of the
Helicteres Isora Panicum virgatum Miscanthus giganteus Humulus lupulus

(Switchgrass) (China reed) (Hop)
Ananas erectifolius Abelmoschus esculentus Abutilon theophrasti Luffa cylindrica

(Curaua) (Okra) (Velvetleaf)
Roystonea regia Phoenix sylvestris Cynara cardunlucus Apium graveolens

(Vakka) (Date) (Artichoke) (Celery)
Figure 3.2 Images of plants considered as new sources of cellulose fibres.
fibres (Figure 3.3b–f).36,112 Thus the dissolution of superficial fatty acids and
lignin components makes the fibres thinner and the pores clearer, enhancing the
ability to graft within a polymer matrix and improving its mechanical prop-
erties.126 For example, the addition of randomly oriented triton-treated isora
fibres to an unsaturated polyester resin was shown to increase the tensile
strength and modulus by about 40% and 140%, respectively, and the flexural
strength and modulus by about 50% and 85%, respectively. The explanation
given for these improvements was the decrease in hydrophilicity brought about
50 Chapter 3
(a) (b) (c)
(d) (e) (f)
Figure 3.3 SEM photographs of (a) untreated, (b) alkali-treated, (c) acylated,
(d) benzoylated, (e) toluene diisocyanate-treated and (f) silane-treated
isora fibres.36,112
by the different treatments, which increases the compatibility of the fibres with
the polyester matrix as well as their dispersion within the polymer.
Switchgrass (Panicum virgatum) is a perennial North American native plant
that requires little water and few nutriments but leads to high productivity.83 It
possesses fibres in its leaves and stems whose mechanical properties and che-
mical composition are interesting enough to foresee various applications.127
For example, owing to their relatively low lignin content, these fibres are quite
easily pulped and thus attractive for paper making industries. Nevertheless,
since switchgrass receives interest mainly as a possible source of bioethanol, few
studies have been considering the possibility of using these fibres in polymer
composites. One study revealed that the incorporation of 30 wt% of switch-
grass fibres to a polypropylene matrix led to an increase by a factor of about 2.5
in the polymer flexural modulus.43
China reed plant, also called Miscanthus, originally comes from Japan, China
and Korea, and is now also harvested in Europe (Figure 3.4).37,128 It takes at
least three years for the plant to mature and to grow several meters high.129 At
that stage, a third of the volume of its stem is made of relatively strong and
straight fibres that can compete, in terms of mechanical properties, with more
traditional vegetable fibres.37 For example, it has recently been shown that
composites made of polypropylene and 30 wt% of China reed fibres or hemp
fibres had the same value of Young’ modulus;128 the addition of such fibres to
the polymer led to increased stiffness by a factor of 2.5. Further enhancement of
the composite properties was achieved by treating the China reed fibres with
polypropylene-grafted maleic anhydride (PP-g-MA): an increase of 12% and
(a) (b) (c)
Figure 3.4 Cross-sections of China reed fibres (F, fibre; P, parenchyma cells;
E, epidermis; L, lumen; M, middle lamella; S1, S2, S3, sub-layers).37,128
(a) (b)
Figure 3.5 SEM micrographs of tensile fracture of PP/30% China reed fibre com-
posite: (a) untreated fibres; (b) fibres treated with 2% PP-g-MA.128
30% of the tensile modulus and the tensile strength, respectively, could be
obtained. Actually, this treatment enables better cohesion between the fibres
and the matrix, as can be seen on composite rupture micrographs (Figure
3.5):128 whereas the untreated fibres are mainly pulled out of the matrix before
the composite failure, the treated fibres are well bonded to the polymer and
the rupture of the composite occurs by fibre breakage as well as fibre slipping.
Attempts have also been made to use Miscanthus fibres to reinforce a
poly(lactic acid) (PLA) matrix: the results of tensile tests showed that this
addition had a positive impact on Young’s modulus (value increased by almost
50%) but a negative impact on strength (decrease of 40%), with or without
addition of compatibilizer.128 However, added to a cellulose diacetate (CDA)
matrix, China reed fibres increased notably both the stiffness and the strength
of the polymer: for a fibre volume fraction of 40%, the composite’s Young’s
modulus and strength were found to be respectively 4 and 1.2 times higher than
the properties of the pure polymer.37 Very similar to China reed plant, elephant
grass (Pennisetum purpureum), originally from Africa, possesses fibres in its
leaves that can be extracted either by chemical or by mechanical means. The
chemical method has been shown to lead to stronger polyester/elephant grass
52 Chapter 3
(a) (b)
Figure 3.6 (a) Optical micrograph showing a cross-section of bundles of okra fibres.
(b) SEM micrograph of a longitudinal view of a bundle of okra fibres.80
composites than the mechanical extraction, with an increase of about 45% of

the tensile strength and modulus.58 Moreover, even the mechanically treated
elephant grass fibres led to an increase of the plain polyester matrix strength
and modulus by a factor of 1.5, for a fibre fraction of 31 vol%.58 These results
were comparable to bamboo/polyester composites.
Okra fibres (Abelmoschus esculentus), also called ‘‘lady’s finger’’ fibres, are
extracted from the stems of a 2 m high plant that mainly grows in Egypt and
Southern Asia. Although it is cropped for its fruits, the mechanical properties
of its bark fibres are similar to those of the more traditional natural fibres used
in composite materials.80 Their colour varies from whitish to yellowish,
depending on the UV radiation received by the plant. Like many other bark
fibres, they are gathered in bundles of several polygonal elementary fibres,
whose thickness varies along the fibre (Figure 3.6).80 Recently, various chemical
treatments have been applied to okra fibres (notably bleaching and acetyla-
tion), aimed at reducing their hydrophilic character.130 While the degradation
of the treated fibres due to exposure to water was greatly reduced compared to
the raw fibres, their mechanical properties dramatically dropped after any of
these treatments. For example, the Young’s modulus of acetylated or bleached
okra fibres was divided by 3 or 4, respectively. However, the effect of adding
treated or untreated okra fibres to polymers has not yet been studied in terms of
mechanical improvements, but it is expected that such fibres would constitute
effective composite reinforcement materials.
Curaua fibres (Ananas erectifolius) are mainly produced in Amazon regions.
The decortication of the leaves provides stiff, strong and smooth bundles of
fibres (Figure 3.7).32,56 Investigations carried out in the last decade on these
fibres as composite reinforcements have already aroused strong interest from
automobile companies.33,131 Actually, a recent study has revealed that the use
of curaua fibres to replace E-glass fibres in certain car parts could lead to
economic, environmental and social benefits, since, for example, they cost 50%
Figure 3.7 Fracture surface of a bundle of curaua fibres.56
less than glass fibres.55 Nevertheless, an effort still has to be made to lighten the
natural fibre composites in order to implement them at a higher level in car
industries. Moreover, alkali treatment has been applied to curaua fibres to
enhance the properties of a cornstarch-based polymer:56 this treatment led to a
drop in the fibres’ properties but also to an improvement in the toughness of the
derived composite. This was attributed to better interfacial bonding induced by
the alkali treatment as well as a reduction of the scattering of fibre strength
values. Also, the addition of 40 wt% of curaua fibres to a polyamide-6 matrix
brought about an increase in the polymer stiffness by a factor of 3, which was
further improved when the fibres were alkali-treated before composite
processing.132
Royal palm (Roystonea regia), also known as vakka, is an Indian and Central
American tree that produces light and strong external fibres.50 These fibres,
extracted from the ripened leaves of the tree, have been recently investigated as
polyester reinforcements.133 The addition of 37 vol% of fibres led to an increase
in the polymer tensile strength and stiffness by a factor of 3, which is higher
than the reinforcement efficiency of other natural fibres such as sisal and
banana (for the same fibre volume fraction). The flexural properties were also
shown to be improved by the presence of vakka fibres, especially the flexural
modulus which increased by a factor of 1.3 for a fibre fraction of 39 vol%.
More surprisingly, the vakka fibre-based composites were shown to exhibit an
increase in their dielectric strength when the fibre fraction increased;133 this acts
in favour of a possible use of such composites in electrical insulation
applications.
The ripe fruit of the Luffa cylindrica, called sponge-gourd, possesses fibres
entangled around the beehive-like structure of the fruit nucleus. This plant is
54 Chapter 3
traditionally harvested in Africa, in Asia and in Central and South America,
either for its edible young fruit or for its mature fruit that can be used as bath
sponges.59,134 Added to the low density of these fibres, their mat structure
makes them worthwhile fibres for polymer reinforcement since they can be used
directly to process composites without the need, for example, to be woven
beforehand. Among the few studies already carried out yet on Luffa fibre-based
composites, one highlighted the beneficial effect of these fibres on the stiffness
of a polypropylene matrix (increase of 40% for a fibre fraction of 15 wt%), in
spite of a reduction in the composite tensile strength compared to the pure
matrix strength.135 Another study confirmed this lack of mechanical
improvement in a polyester matrix reinforced with Luffa fibres, but underlined
the modification to the fracture mode of the polymer induced by this addition
(from a fragile one to a more controlled one), making these fibres attractive in
hybrid composites.136
Date (Phoenix sylvestris) is a palm tree from the South East of Asia that
possesses fibres both in the stems of its leaves and in the netted structure that
surrounds the base of the tree (called amplexicaul fibres). The first type of fibre is
extracted by beating the leaves followed by water retting, whereas the second
type is obtained by carding the dried netted structure. Even if they differ a lot in
terms of stiffness, these two types of fibres are relatively light and strong and
could be used to strengthen parts of polymer composites.50 Added to an epoxy
matrix, date fibres extracted from leaves increased the flexural modulus by about
50% and even by 90% when the fibres were treated beforehand with acetic and
maleic anhydrides.78 This enhancement was much lower in the case of an
unsaturated polyester matrix (13%) and was even null with treated fibres.
Moreover, date palm fibres were shown to enhance the thermal and mechanical
stabilities of a polypropylene matrix submitted to severe weathering condi-
tions.137 This is a great advantage when dealing with outdoor uses of composites.
Hop (Humulus lupulus) is a plant mainly produced in the USA, in Germany
and in Ethiopia. It belongs to the same family as hemp (Cannabaceae) and is
thus a potential source of cellulose fibres, since its stem, although generally
considered as a by-product, possesses external and internal fibres.35 The surface
of a hop stem is irregular and coated with numerous deposits of non-cellulosic
substances, most of which are removed during fibre extraction, leading to
smooth and clean bundles of fibres (Figure 3.8). A recent study revealed that
the addition of fibres extracted from the outer bark of a hop stem to a poly-
propylene matrix led to better mechanical properties than jute fibres, for the
same composite density.138
Rhectophyllum fibres (Rhectophyllum camerunense) are plant fibres that can
be extracted from the long air roots of a common creeper native to the Gulf of
Guinea.41 These bundles of fibres are very long and porous and exhibit a very
low density. Their high cellulose content and good mean specific properties are
likely to make them attractive as reinforcement materials for composites, even
if they have not yet been tested in this role.
Like nettle, velvetleaf is often considered as a weed since this plant (Abutilon
theophrasti) thrives to the detriment of other crops. This invasiveness, added to
(a) (b)
Figure 3.8 SEM images of (a) a hop stem and (b) a bundle of hop fibres.35
mechanical properties very close to those of more common natural fibres, as

well as the recent improvement of fibre processing, acts in favour of using these
fibres as polymer reinforcements.45 Nevertheless, no study has yet been carried
out to determine the properties of the derived composites.
Also considered as by-products, corn and artichokes stems, cotton stalks,
corn and rice husks, rice and soybean straws have all been shown to contain the
same kind of ligno-cellulosic fibres as those mentioned above.19,23,25,31,42,82,139
Moreover, as revealed by SEM images (Figure 3.9), the bundles extracted from
these products also have the same type of morphology as the traditional plant
bundles, i.e. rough elementary fibres glued together in scores by non-cellulosic
components. They are thus likely to provide effective composite reinforcement,
although the effect of most of them has not yet been characterized. The main
conclusion of the studies carried out on these by-products is that their wide
availability and their low cost compensate for their relatively low mechanical
properties. What can also be noted is the similar shape between the fibres
extracted from some of these by-products and cotton fibres, with several con-
volutions along the length of elementary fibres (Figure 3.10).25,42,45,140
Among the different forms of plant waste, rice straws have been largely
studied these last few years as potential polymer reinforcements. For example,
incorporating 50 wt% of rice straw fibres into a high-density polyethylene
matrix was shown to increase the storage modulus, tensile strength and impact
strength of the polymer by about 150, 50 and 25%, respectively.139 Another
example concerns rice straw/polyester composites made of 40 vol% of fibres:
their flexural strength and modulus were 20 and 54% higher than those of
the plain matrix.141 More recently, composites made of poly(lactic acid) and
20 wt% rice straw fibres treated with 8 wt% poly(butyl acrylate) were tensile
tested and exhibited a tensile strength 25% higher than that of the neat
matrix.142 Added to waste tyres, rice straw fibres led to composites that pre-
sented higher flexural properties and acoustic insulation than wood particle
boards, giving a foretaste of these cheap natural fibres as effective reinforce-
ments of recycled polymers in insulation boards.143
56 Chapter 3
(a) (b) (c)
(d) (e) (f)
Figure 3.9 SEM images of untreated bundles of fibres extracted from (a) artichoke
stem,23 (b) corn stalk,31 (c) corn husk,19 (d) rice straw,82 (e) soybean
straw42 and (f) cotton stalk.25
(a) (b) (c) (d)
Figure 3.10 SEM images of elementary fibres of (a) soybean straw,42 (b) cotton
stalk25 and (c) velvet leaf,45 having the same convolutions as seen in
(d) cotton.140
3.4 Concluding Remarks

Despite the numerous hindrances related to natural fibres, notably the strong
dependence of their mechanical behaviour on temperature and humidity and
their lack of reliability, promising developments of plant fibre composites are
expected since ecological concerns are growing relatively fast in many domains.
Current studies tend, among others, to limit the influence of external conditions
on the composites’ stability in order to enhance the reliance of industrialists on
such materials, while other investigations aim at increasing their mechanical
properties to improve their use and popularity among composite users. One
particular objective is to substitute synthetic fibres, mainly E-glass fibres, by
natural fibres, which could help in improving the carbon footprint and the
energy balance for the production of such eco-composites.
Improvements in the reliability of natural fibre based composites, i.e.
reduction of the fibres properties ranges, could be achieved by different means,
such as a better fibre selection. This could be done, for example, by improving
the fibre extraction processes, by selecting varieties or even by modifying plants
genetically in order to obtain almost identical and defect-free fibres, for a given
harvesting area. Nevertheless, this last idea, still under debate in relation to
other natural products (especially food resources), is far from being the most
‘‘environmentally friendly’’ solution.
Hopefully, as the number of scientific papers dealing with the characteriza-
tion of new plant fibres for polymer reinforcement is increasing every month,
the development of natural fibre-based composites is expected to increase,
especially at a local level, thanks to the large variety of species available
worldwide. Likewise, the use of materials previously considered as by-products
or waste, to strengthen or stiffen polymeric matrices, is a fashionable, popular
and certainly profitable way to respond to government expectations in com-
posite applications.
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2010, 41, 588.
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Bioresour. Technol., 2004, 95, 61.
CHAPTER 4
Relation between Structural

Anisotropy in Natural Fibres
and Mechanical Properties in
Composites
ELESSANDRA DA ROSA ZAVAREZE*a AND
ALVARO RENATO GUERRA DIASb
a
School of Food Engineering, Federal University of Pampa, 96413-170,
Bagé, Brazil; b Department of Agroindustrial Science and Technology,
Federal University of Pelotas, 96010-900, Pelotas, Brazil
4.1 Introduction
As a result of the increasing demand for environmentally friendly materials and
the desire to reduce the cost of traditional fibres (carbon, glass and aramid) and
reinforced petroleum-based composites, new bio-based composites have been
developed.1 Researchers have begun to focus their attention on natural fibre
composites, which are composed of natural or synthetic resins, reinforced with
natural fibres. The natural fibre component may be wood, sisal, flax, hemp,
coconut, cotton, flax, jute, banana leaf fibres, bamboo, wheat straw or other
fibrous material. Natural fibres can be divided into vegetable, animal and
mineral fibres. The main polymers involved in the composition of vegetable
fibres are cellulose, hemicelluloses, lignin and pectin, while fibres of animal
origin consist of proteins (hair, silk, wool).2

63
64 Chapter 4
Polymer matrix composites using natural plant fibres as reinforcement have
attracted much interest in recent years. This is because these natural fibres can
be harvested from renewable resources, possess long aspect ratios for efficient
stress transfer, and certain aspects of their mechanical properties are compar-
able to those of existing inorganic glass fibres.3 Thermoplastics are alternative
matrix materials that possess a number of clear advantages over thermoset
matrices. These include enhanced shelf-life, post-forming, toughness and
recyclability. The physical and chemical nature of a fibre surface defines its
compatibility and adhesion with a polymer, as well as determining such
characteristics as surface free energy, wettability and the possibility of inter-
action with the matrix or coupling agents. Natural fibres offer many advantages
such as energy efficiency, low cost, low density, high toughness, acceptable
specific strength and bio-degradability.4,5 However, they do suffer from a few
limitations because bio-fibres are hydrophilic in nature and thus are less
compatible with relatively hydrophobic polymer matrices. The hydrophilic
nature of bio-fibres is also responsible for the water absorption characteristics
of biocomposites, thus limiting their application. Because the mechanical
properties of composites are related to the compatibility and interaction
between their components, the improvement of the interface and interphase
interactions in natural fibre/polyester composites is essential.6
Fibres serve as reinforcement in composites and show high tensile strength
and stiffness, while the matrix holds the fibres together, transmits the shear
forces and also functions as a coating. The selection of suitable fibres is
determined by the required values of stiffness and tensile strength of a
composite. Further criteria for choosing suitable reinforcing fibres include
elongation at failure, thermal stability, adhesion of the fibres and matrix,
dynamic behaviour, long-term behaviour and price and processing costs.2 The
mechanical properties of fibres determine the stiffness and tensile strength of a
composite. Depending on the fibre orientation, the materials behaviour of
composites can be divided into quasi-isotropic (with all short fibres randomly
orientated, and no privileged direction of mechanical properties), anisotropic
(with all fibres orientated in one or more directions with corresponding
mechanical properties) or orthotropic (fibres orientated mainly in two
directions orthogonal to each other and showing corresponding materials
behaviour).2,7 Owing to the anisotropy in composites reinforced with natural
fibres, some properties modify the fibre orientation, i.e. the mechanical prop-
erties depend on the direction in which force is applied. The fibres adopt a
particular orientation in the process, which is responsible for the increased
stiffness in the direction parallel to the flow. Materials can be classified as either
isotropic or anisotropic. Isotropic materials have the same material properties
in all directions, and anisotropic materials have different properties along
different directions with respect to a specified point in a material.7
The mechanical properties of composite materials depend on the structure in
a complex way. The mechanical properties of natural fibres are affected by the
variability in plants and by the processing stage and damage sustained during
processing; thus, there is a large distribution in observed mechanical
Structural Anisotropy in Natural Fibres and Mechanical Properties in Composites 65
8,9
properties. The mechanical performance of composites is mainly dependent
upon the properties of the matrix and reinforcement material and the interaction
between the matrix and reinforcement. Moreover, the chemical composition,
crystallinity, surface properties, diameter, cross-sectional shape, length, strength
and stiffness vary from fibre to fibre. Natural fibres can improve the mechanical
properties of composites, particularly thermoplastic products. Fibre-reinforced
plastic composites are used in several structural applications and fields such as
construction, aerospace, automotive parts, sports equipment, machinery and the
food industry.10
Many researchers have studied the effect of natural fibres on the properties of
reinforced composites.2,3,6,8,11–24 This chapter will review the properties of
natural fibres and composites, emphasizing the co-relation between the struc-
tural anisotropy in natural fibres and the mechanical properties in composites.
The importance and applications of natural fibre composites will also be
discussed.
4.2 Natural Fibres

Natural fibres can be further subdivided into vegetable, animal and mineral
fibres. Mineral fibres are generally used in very small amounts, often due to
factors that affect human health during extraction. Vegetable fibres (e.g., cotton,
flax, hemp, jute) are composed of cellulose, while fibres of animal origin consist
of proteins (e.g., hair, silk, wool). The surfaces of natural fibres are rough and
uneven and provide good adhesion to the matrix in a composite structure. When
determining the properties of natural fibres, one has to keep in mind that one is
dealing with natural products with properties that are strongly influenced by
their natural growth environment, such as temperature, humidity, the compo-
sition of the soil, strength of its fibres, density, etc. Additionally, the way the
plants are harvested and processed results in a variation in properties.25
Natural fibres are rounded, elongated structures with hollow cross-sections
spread over the entire plant and are classified according to the anatomic origin
of the stem, leaf, wood and surface fibres. Stem fibres occur in the phloem of,
for example, jute, ramie, flax and palm fibres. Banana is also extracted from the
pseudo-stem of the plant. Leaf fibres are extracted from the leaves of plants
such as sisal, curaua, pineapple and palm. Wood fibres can be subdivided into
two types, hardwood and softwood, and these have different properties. Wood
is a hydrophilic porous composite of cellulose, lignin and hemicellulose poly-
mers that are rich in functional groups such as hydroxyls, which readily interact
with water molecules by hydrogen bonding.26 Surface fibres form a protective
layer on the stems, leaves, fruits and seeds of plants such as açaı́, coconut and
cotton. Vegetable fibres are composed primarily of cellulose, hemicellulose,
lignin and small amounts of pectin, inorganic salts, nitrogenous substances and
natural dyes, which constitute what is called the soluble fraction. Cellulose, the
largest fractional component in vegetable fibres, is a semicrystalline poly-
saccharide made up of D-glucosidic bonds. The large number of hydroxyl
66 Chapter 4
groups in cellulose (three in each repeating unit) imparts hydrophilic properties
to natural fibres. Hemicellulose is strongly bound to the cellulose fibrils, pre-
sumably by hydrogen bonds. Hemicellulose polymers are branched and fully
amorphous and have a significantly lower molar mass than cellulose. Because
of its open structure containing many hydroxyl and acetyl groups, hemi-
cellulose is partly soluble in water and is hygroscopic. Lignins are amorphous,
highly complex, mainly aromatic polymers with phenylpropane units, but have
the lowest water sorption of natural fibre components.27,28
A fibre has a length that is much greater than its diameter. The length-to-
diameter ratio is known as the aspect ratio, which can vary greatly. Continuous
fibres have high aspect ratios, whereas discontinuous fibres have low aspect
ratios. Continuous-fibre composites normally have a preferred orientation,
while discontinuous fibres generally have a random orientation.7 Each indivi-
dual fibre cell has a complex structure formed by cell walls that surround the
lumen. The walls consist of layers formed by clusters of microfibrils in plants
that grow in a spiral shape with different angular orientations. The main
component of microfibrils is cellulose; cellulose microfibrils are interconnected
through a network of hemicellulose molecules. Another important constituent
of the walls is lignin, a primarily hydrophobic substance that forms an
impregnated layer near the surface, which is responsible for support. Micro-
fibrils are composed of micelles, which are crystals of cellulose molecules
arranged parallel to different allotropic structures.
4.3 Sources of Natural Fibres

Plant fibre sources include cotton, flax, hemp, sisal, jute, kenaf, henequen, corn
and coconut. Fibres from mineral sources include asbestos fibres. Asbestos is
the generic commercial designation for a group of naturally occurring mineral
silicate fibres of the serpentine and amphibole series. These include the
serpentine mineral chrysotile (white asbestos) and the five amphibole minerals:
actinolite, amosite (brown asbestos), anthophyllite, crocidolite (blue asbestos)
and tremolite. Asbestos consist of short, discontinuous fibres of high length/
diameter ratio and high surface area. Fibres from animal sources include silk
and wool. Fibres from animal sources include wool, a fibre formed in the fleece
of sheep or similarly hairy animals (e.g., alpacas, llamas, vicunas, yaks, camels,
cashmere goats, mohair goats and angora rabbits) produced in many places
around the world. After shearing, the wool is washed, carded and dyed before
being woven or knitted into fabric. Wool is flexible and absorbs moisture,
making it cool in summer and warm in winter. Silk is another fibre from animal
sources that is produced from the cocoons of the silkworm. Silk absorbs
moisture, dries quickly, will not shrink, is easily dyed, retains its shape and has
a natural shimmer.
Natural fibres exhibit a variation in properties due to environmental con-
ditions. There are many types of vegetable fibres, which are the most important
and most used fibres in several industrial applications.
4.3.1 Cotton Fibre
Worldwide, cotton is one of the most important fibres used in the textile
industry. Hand picking of cotton fruits are highly labour-intensive, which is
often carried out by machine. In many parts of the world, however, picking is still
carried out by hand. The seeds, dead leaves and other debris are removed by
ginning. The cotton can absorb up to 20% of its dry weight in moisture without
feeling wet and is also a good heat conductor.
4.3.2 Jute Fibre

According to the International Jute Study Group,29 jute is one of the most well-
known vegetable fibres, which is mostly grown in countries such as India,
Bangladesh, China, Nepal and Thailand. Together, they produce about 95% of
the global production of jute fibres. Like those of synthetic fibre composites, the
mechanical properties of the final products made of jute fibres also depend on
the individual properties of the matrix, fibre and the nature of the interface
between them.19 The fibres of jute are extracted from the ribbon of the stem.
These fibres are obtained by successively retting in water, beating, stripping the
fibre from the core and drying. Owing to its short fibre length, jute is the
weakest stem fibre, although it withstands rotting very easily. It is used as a
packaging material and in carpet backing, ropes, yarns and wall decorations.25
4.3.3 Flax Fibre

Flax fibre is readily marketed in a variety of industries, including textiles,
composites and specialty paper. The individual fibres are formed in bundles
that encircle the core tissue. Flax must undergo the process of retting to
separate the fibre from woody cells, which are termed shives and constitute the
major waste component of flax fibres.30 When ripe, flax plants are pulled from
the ground rather than cut, to avoid a loss in fibre length from the stubble left in
the field. Flax is a strong fibre and experiences an increase in strength under wet
conditions and can absorb 20% moisture without feeling wet. Flax has good
heat conducting properties and is hard wearing and durable. Flax is used in the
production of linen, ropes and sacks.
4.3.4 Ramie Fibre

Ramie produces some of the strongest and longest plant fibres, which are
lustrous with an almost silky appearance. Fibres from ramie are used in
clothing fabrics, industrial packaging, twines, cordages, canvas, car outfits, etc.
Recently, ramie fibres were successfully tested in fibre-reinforced composites.31
4.3.5 Hemp Fibre

Hemp fibres are present in bundles that are as long as the stems, which can
easily be peeled off the xylem surface by hand or machine. The fresh stem
68 Chapter 4
consists of a hollow cylinder of 1–5 mm thick xylem covered by 10–50 mm
cambium, 100–300 mm cortex, 20–100 mm epidermis and 2–5 mm cuticle.14 A
hemp yarn is strong and has the highest resistance against water of all natural
fibres; however, it should not be creased excessively, to avoid breakage. Fibres
from hemp stems have been widely used in the production of cords and clothing
and have potential as a reinforcement material in polymer/matrix composites.
4.3.6 Sisal Fibre

Sisal fibre is one of the most widely used natural fibres and is very easily cul-
tivated. Sisal fibre is a hard fibre extracted from the leaves of the sisal plant
(Agave sisalana), which originated in Mexico and is now mainly cultivated in
East Africa, Brazil, Haiti, India and Indonesia. A sisal plant produces about
200–250 leaves, each of which contains 1000–1200 fibre bundles composed of
4% fibre, 0.75% cuticle, 8% dry matter and 87.25% water.32 Sisal produces
sturdy and strong fibres that are very well resistant against moisture and heat. It
is mainly used in ropes, mats, carpets and as a cement reinforcement.25 The
characteristics of the sisal fibres depend on the properties of the individual
constituents, the fibrillar structure and the lamellae matrix. Sisal fibres are
composed of numerous elongated fusiform fibre cells that taper towards each
end. The fibre cells are linked together by middle lamellae, which consist of
hemicellulose, lignin and pectin.
4.3.7 Henequen Fibre

Henequen fibre (Agave fourcroydes) is composed of approximately 77%
cellulose, 4–8% hemicelluloses, 13% lignin and 2–6% pectin and waxes by
weight. Henequen has been used extensively to make ropes, carpets and cor-
dages for a long period of time.33 The structural arrangement of lignin and
cellulose inside the fibre is based fundamentally on lignin, which acts as a
cementing matrix for the cellulose component and has its own separate struc-
ture. The cellulose fibre is the rigid part of the fibre and the mechanical
properties of henequen fibre such as stiffness and strength are mainly influenced
by its structural arrangement.34
4.3.8 Coir Fibre

Coir is a natural fibre extracted from the husk of coconut and used in products
such as floor mats, doormats, brushes and mattresses. Coconut fibre is obtained
from the husk of the fruit of the coconut palm. The fruits are dehusked on a
spike, and after retting, the fibres are extracted from the husk by beating and
washing. The fibres are strong, light and can easily withstand heat and salt
water. After nine months of growth, the nuts are still green and contain white
fibres, which can be used for the production of yarn, rope and fishing nets.
After 12 months of growth, the fibres are brown and can be used for brushes
and mattresses.25
4.4 Composites
4.4.1 Biopolymer Composites
A composite material can be defined as a combination of two or more materials
that results in better properties than those of the individual components used
alone. Unlike metallic alloys, each material retains its individual chemical,
physical and mechanical properties. The two constituents behave as a rein-
forcement and a matrix, respectively. The main advantages of composite
materials are their high strength and stiffness combined with low density,
compared with bulk materials, allowing for a weight reduction in the finished
product.7
Biodegradable polymers are designed to decompose upon disposal by the
action of living organisms. Extraordinary progress has been made in the
development of practical processes and products using polymers such as starch,
cellulose and lactic acid.35 Biopolymer composites are becoming increasingly
employed to replace oil-based components.36 Understanding their mechanical
properties is essential to guarantee high performance during their lifetime.13
The main impediment to the development of biopolymers is their low
mechanical performance and high sensitivity to environmental conditions
(temperature, humidity).37
According to Guessasma and Bassir,38 the use of biopolymer-based com-
posites such as starchy materials to replace oil-based materials is a challenging
task that has led to several recent research developments. The improvement in
the mechanical performance of biocomposites is a key prerequisite to increase
the potential of their use as materials in packaging and single-use components,
among other applications. The improvement in the mechanical performance of
thermomoulded glassy biocomposites is correlated to the basic understanding
of the relationship between the composites’ elastic properties and micro-
structural attributes. Recently, the tailoring of the elastic properties of such
composites has been proved to be dependent on the composites’ intrinsic
properties, the spatial arrangement of phases and interface behaviour.36 Starch
is an inexpensive, annually renewable material derived from corn and other
crops. The biodegradation of starch products recycles atmospheric CO2 trap-
ped by starch-producing plants. The starches contain amylose and amylopectin
at ratios that vary with starch source. This variation provides a natural
mechanism for regulating the material properties of starch.35
Biopolymers such as cellulose, starch and proteins can also be modified to
produce composites. Chemically modified plant cellulose is used in a remark-
ably diverse set of applications. For example, cellulose acetate is used in many
common applications, including toothbrush handles and adhesive tape back-
ing.35 Kadokawa et al.16 studied the properties of cellulose/starch composite
fibrous material by reconstitution from a mixture. These authors indicate that
the crystalline structures of the cellulose and starch were largely disrupted in the
gel, and the scanning electron microscopy and X-ray diffraction data of the
obtained material show a compatibilized fibrous structure. The cellulose and
70 Chapter 4
starch are composed of the same unit structure, i.e. a glucose unit, but are
linked through different glycosidic bonds, such as b-(1-4) for cellulose and
a-(1-4) and a-(1-6) for starch. The roles of cellulose and starch in nature are
completely different; the former is a structural material and the latter is an
energy storage material. Cellulose is the most abundant organic substance on
Earth and, thus, a very important renewable resource that has a number of
traditional applications, including its use in furniture, clothing and medical
products.39
Kumar and Zhang17 studied the properties of aligned ramie-fibre-reinforced
and soy protein composites and described that the alignments of fibres along
different directions had a significant effect on the mechanical properties of
the soy protein/ramie fibre composites; moreover, the soy protein/ramie fibre
composites with vertically aligned fibres exhibited the highest mechanical
properties. The arylation of the soy protein/ramie fibre composites led to a
significant improvement in water resistance and elastic modulus. The mechanical
properties of the protein composites are proportional to the volume fraction of
fibres as well as the orientation of the fibres. To improve the properties of soy
protein isolate materials, a series of modified protein composites such as soy
protein isolate thermoplastic reinforced with chitin whiskers,40 montmorillonite
exfoliated soy protein isolate nanocomposites41 and water-induced hydrophobic
soy protein isolate materials42 have been reported.
4.4.2 Thermoplastic Composites

Thermoplastics are polymers that require heat to make them processable. After
cooling, such materials retain their shape. In addition, these polymers may be
reheated and reformed, often without significant changes in their properties.43
Thermoplastic polymers constitute an important class of materials with a wide
variety of applications. Thermoplastic composites offer some substantial
advantages over thermoset composites. Owing to the higher toughness of the
matrix, they offer a higher impact resistance. Manufacturing cycle times
consisting of melting the matrix, shaping and consolidation by cooling are
significantly shorter than those for thermosets, which require a time-consuming
curing step. The main disadvantage of thermoplastic composites is the need for
high processing temperatures and pressures, caused by the high melt viscosity
of the matrix. In addition, proper impregnation of the fibres at the micron level
has proven difficult and often results in products with a locally high void
content. One way to improve fibre impregnation is to bring the matrix and the
fibres in more intimate contact before the final moulding step or, in other
words, to reduce the required flow length of the polymer matrix.44,45 According
to Riedel and Nickel,2 the impregnation of fibres in a composite structure,
particularly thermoplastic resins, depends on the viscosity of the matrix, which
can be influenced by temperature.
Polymers of natural origin (e.g., starch and cellulose) must be modified either
physically or chemically to make them suitable for processing as thermoplastic
resins. For example, the structure of starch can be made thermoplastic with
adjuvants such as glycerol and water. The physical, chemical, mechanical and
thermal properties of biopolymers are also influenced by physical and chemical
modifications. The esterification of hydroxyl groups on side chains is preferred to
convert cellulose into a thermoplastic material that retains its cellulose chain
structure.2 Starch polymers have been applied in several areas, including in star-
ches used in composites, building materials and polymer blends. The main
drawbacks of thermoplastic starches are their brittleness, moisture absorption and
low mechanical properties. The incorporation of natural fibres is an alternative to
improve their thermal and mechanical properties and preserve their biodegrad-
ability. Lignocellulosic fibres can be used for the development of new composites
because of their good mechanical and physical properties.23
Ma et al.46 reported on the properties of thermoplastic starch composites
with micro winceyette fibre as reinforcement. The introduction of micro
winceyette fibre improved the tensile strength, water resistance and thermal
stability of thermoplastic starch because of the good adhesion between the
starch and fibre. With the increase in fibre content from 0 to 20%, the initial
tensile strength was trebled up to 15.16 MPa, while the elongation was reduced
from 105 to 19%. The Young’s modulus of thermoplastic starch behaved
analogously to the tensile strength as a consequence of introducing fibres. The
reinforcement effect was gradually weakened with the increase in water content,
but at high water content (430%) both the fibre and water content had
no effect on the tensile strength. A considerable increase in tensile strength
indicated that thermoplastic starch was well suited as the matrix for natural
cellulose fibres. These authors reported that this was due to the remarkable
intrinsic adhesion of the fibre/matrix interface caused by the chemical similarity
of starch and the cellulose fibre.
4.4.3 Natural Fibre Composites

Today, natural fibres constitute an interesting alternative in composite
technology. The use of fibres such as flax, hemp, jute or sisal in this industry to
date is small, in part because the availability of a durable semi-finished product
with consistently good quality is often a problem. The natural fibres are
embedded in a biopolymer matrix system, the function of which is to hold the
fibres together. This provides and stabilizes the shape of the composite struc-
ture, transmits the shear forces between the mechanically high-quality fibres,
and protects them against radiation and other aggressive media.2
The properties of composites are strongly dependent on the properties of
their constituent materials, their distribution and the interaction between them.
The composite properties may be the volume fraction sum of the properties of
the constituents, or the constituents may interact in a synergistic way, resulting
in improved or better properties. Apart from the nature of the constituent
materials, the geometrical properties of the reinforcement, such as shape, size
and size distribution, influence the properties of the composite to a great extent.
The concentration distribution and orientation of the reinforcement also affect
72 Chapter 4
the properties. The shape of the discontinuous phase, which may by spherical,
cylindrical or rectangular cross-sanctioned prisms or platelets, the size
distribution, which controls the texture of the material, and volume fraction
determine the interfacial area; this area plays an important role in determining
the extent of the interaction between the reinforcement and the matrix.18
Although natural fibres can offer the resulting composites many advantages,
typically, polar fibres have inherently low compatibility with non-polar poly-
mer matrices, especially hydrocarbon matrices such as polypropylene and
polyethylene.47 Xie et al.48 reported that this incompatibility may cause pro-
blems during composite processing and for the material properties. Hydrogen
bonds may form between the hydrophilic fibres, and thus the fibres tend to
agglomerate into bundles and unevenly distribute throughout the non-polar
polymer matrix during compounding processing.
Research and development have shown that these aspects can be improved
considerably. Because natural fibres are cheap and have a better stiffness per
weight than does glass, which results in lighter components, interest in natural
fibres has grown. Moreover, the environmental impact is smaller because the
natural fibre can be thermally recycled and fibres come from a renewable
resource. Their moderate mechanical properties keep fibres from being used in
high-tech applications, but for many reasons they can compete with glass fibres.
The advantages are as follows: (1) low specific weight, which results in a higher
specific strength and stiffness than glass; (2) it is a renewable resource and their
production requires little energy; (3) they can be produced with low investment
at low cost; (4) the feature friendly processing, with no wear of tooling or skin
irritation; (5) thermal recycling is possible, whereas glass causes problems in
combustion furnaces; and (6) good thermal and acoustic insulating properties.
The disadvantages are as follows: (1) lower strength properties, particularly
impact strength; (2) variable quality, depending on unpredictable influences
such as weather; (3) moisture absorption, which causes swelling of the fibres;
(4) restricted maximum processing temperature; (5) lower durability, though
fibre treatments can improve this considerably; (6) poor fire resistance; and
(7) the price can fluctuate by harvest results or agricultural politics.25
The properties of composite materials depend very much upon their
structure. Composites differ from homogeneous materials in that considerable
control can be exerted over larger-scale structures and, hence, over the desired
properties. In particular, the properties of a composite material depend upon
the shape of the heterogeneities, upon the volume fraction occupied by them
and upon the interface among the constituents. The shapes of the hetero-
geneities in a composite material are classified as follows. The principal
inclusion shape categories are: (1) the particle, with no long dimension; (2) the
fibre, with one long dimension; and (3) the platelet or lamina, with two long
dimensions. The inclusions may vary in size and shape within a category. For
example, particulate inclusions may be spherical, ellipsoidal, polyhedral or
irregular. If one phase consists of voids, filled with air or liquid, the material is
referred to as a cellular solid. If the cells are polygonal, the material is a
honeycomb; if the cells are polyhedral, it is a foam. In each composite structure
we may moreover make the distinction between random orientation and pre-
ferred orientation.49
Natural fibres are being widely used in fibre-reinforced polymer composites.
A common feature of natural fibres is a much higher variability of mechanical
properties. This necessitates study of the strength distribution of the fibres and
efficient experimental methods for its determination. Common fibre reinforced
composites are composed of fibres and a matrix. The fibres are the reinforce-
ment and the main source of strength, while the matrix glues all the fibres
together in shape and transfers stresses between the reinforcing fibres. The
fibres carry the loads along their longitudinal directions. Sometimes, filler
might be added to smooth the manufacturing process, impact special properties
to the composites and/or to reduce the product cost.18 Fibre-reinforced com-
posites require a moderate adhesion between the matrix and fibre. Good
adhesion between the two phases provides good mechanical strength by effi-
cient transfer of load from the matrix to the fibre, but the material becomes
brittle. Poor adherence results in low mechanical strength, but the energy
absorbed increases the fracture energy dissipation during the detachment of the
fibre. Researchers have investigated applications from several natural fibres,
such as hemp,14 flax,9 sisal,12,21 oil palm,10 jute15 and cotton fibres20,50 to
improve the properties of composites.
4.4.4 Anisotropy
The material behaviour of composites depends on the fibre orientation; when
the material is anisotropic, all fibres are orientated in one or more directions,
with corresponding mechanical properties.2,7 The anisotropy of materials is
defined as the variation in the material’s physical response to the applied stress
along different specimen axes. The local average fibre direction and the degree
of anisotropy can be evaluated based on parameters related to the distribution
of the different phases in a representative volume, without the need to analyse
and/or reconstruct the geometry of each single fibre.51 The fibre length dis-
tribution in a material sample can be easily determined by separating the fibres
form the matrix by burning or hydrolysis and observing them using an optical
microscope, although further fibre breakage during extraction cannot be
excluded. The measurement of fibre orientation is more difficult because fibres
are dispersed in the matrix and cannot be separated from the matrix without
altering their orientation.48 Consequently, simplifying assumptions with respect
to fibre orientation are usually made to apply these mechanical models, and
fibre orientation factors are derived implicitly from tests rather than evaluated
on the basis of fibre angles measurement.52 Lillie et al.53 also reported that it is
important to determine the inherent anisotropy due to certain structural
features of an unstressed material, such as an unequal partitioning of ortho-
gonal fibres and induced anisotropy due to superimposed stresses on a non-
linear material in which elastic moduli depend on loading conditions.
Anisotropic properties can be obtained by the addition of aligned organic or
inorganic fibrous fillers. The incorporation of short fibres has caused many
74 Chapter 4
technical difficulties, such as an increase in the viscosity of the blend system,
machine wear problems and increased energy consumption.27 The fibre length
distribution and fibre orientation distribution in composites play an important
role in determining the composite mechanical properties. Owing to the partial
fibre orientation, short-fibre composites can exhibit more or less anisotropy
with respect to their mechanical properties. This inevitability of anisotropy in
the mechanical properties of short-fibre reinforced composites can lead to
severe deterioration in the total performance of the composites when their
unfavourable direction is highly loaded or under attack by the environment.54
Michalowski and Cermák55 tested three distributions of fibres: random orien-
tation, all fibres in the vertical direction and all fibres in the horizontal direc-
tion. These authors reported that the contribution of the fibres to the composite
strength was the largest when they were placed in the direction of largest
extension of the composite (here, horizontal). Vertical fibres in triaxial testing
were subjected to compression; they had an adverse effect on the initial stiffness
of the composite and they did not contribute to any strength increase. Speci-
mens with a random distribution of fibres exhibited a smaller increase
in strength than those with horizontal fibres because a portion of randomly
distributed fibres was subjected to compression.
Fibre-reinforced composite materials typically exhibit anisotropy. That is,
some properties vary depending upon which geometric axis or plane they are
measured along. For a composite to be isotropic with respect to a specific
property, such as Young’s modulus, all reinforcing elements, whether fibres or
particles, must be randomly oriented. This is not easily achieved for dis-
continuous fibres because most processing methods tend to impart a certain
orientation to the fibres. Continuous fibres in the form of sheets are usually
used to deliberately align the composite anisotropy along a particular direction
that is known to be the principally loaded axis or plane. Honda and Narita56
focused their study on locally anisotropic structures of fibrous laminated
composites and reported that they are often found as parts of natural com-
pounds and that the bony pelvis of humans is composed of inorganic calcium
phosphates and organic collagen fibres. The inorganic component contributes
to stiffness and strength, while organic fibres provide toughness. The organic
fibres form curvilinear shapes, which may be the optimum shape to sustain
external forces. In other words, the natural compounds have locally anisotropic
properties distributed optimally throughout the compound to perform more
effectively than simple anisotropic materials.
To develop structure–property relationships in composite systems, it is
necessary to know the properties of the fibre and matrix constituents. Much
research indicates that natural fibres exhibit complicated anisotropic structure.
Natural fibres are often composed of millimetre-long microtubules generally
oriented along the fibre direction. These microtubules are in turn composed of a
variety of substances, including cellulose, lignin and hemicelluloses. The cel-
lulose forms crystalline fibrils bound to one another by non-crystalline cellulose
that spiral within the microtubules along the general direction of the natural
fibre axis. Models indicate that fibre stiffness is influenced by the spiral angle of
the crystalline fibrils as well as the concentration of the non-cellulosic
substances. These structural parameters vary between the different types of
natural fibres, likely accounting for some of the variations in reported fibre
properties.32,57
Demirci et al.58 reported that the random orientation of fibres is the main
source of mechanical anisotropy, and the orientation distribution function of
the fibrous matrix can be used to analyse the structure’s direction-dependent
behaviour. Besides, a practical tool is developed to determine an orientation
distribution function and anisotropic parameters based on its images obtained
with scanning electron microscopy or X-ray micro-computed tomography
regardless of its planar density.
4.4.5 Mechanical Properties

Tensile strength, tensile modulus and elongation at break of the composites
vary as the properties and composition of the fibre and characteristics of
the matrix of the composite. The specific mechanical properties of natural
fibres are important when used in composites. When comparing the tenacity
and elongation at failure of both natural and synthetic fibres, hemp, flax and
ramie fibres can compete with glass fibres, which serve as a reference because of
their major importance in composite technology.2 According to John and
Thomas,1 the primary effects of biofibre reinforcement on the mechanical
properties of natural rubber composites include increased modulus, increased
strength with good bonding at high fibre concentrations, decreased elongation
at failure, greatly improved creep resistance over particulate-filled rubber,
increased hardness and a substantial improvement in cut, tear and puncture
resistance.
Both the long-term and short-term mechanical performance of a natural fibre
composite depend on the properties of the constituents, their relative volume
fractions and on their microscopic architecture, for example, the fibre orien-
tation distribution, the fibre length distribution, the fibre shape, the size of
lumen and whether it is filled with resin or not. Other aspects of equally high
importance are variable fibre parameters such as surface morphology and
surface composition.59 The mechanical properties of a fibre-reinforced polymer
composite depend not only on the properties of the constituents but also on the
properties of the region surrounding the fibre, known as the interphase. Stress
transfer from the matrix to the fibre takes place at such an interphase and,
therefore, it is important to characterize its properties to better understand the
performance of the composite. Sreekala et al.10 studied the tensile strength,
tensile modulus and elongation at break of the hybrid composites of oil palm
fibre hybridized with glass fibre to achieve superior mechanical performance.
They reported that hybrid composites are materials created by combining two
or more different types of fibres in a common matrix. These authors also
reported that the mechanical performance of composites is mainly dependent
upon the properties of the matrix and the reinforcement and the interaction
between the matrix and the reinforcement.
76 Chapter 4
The mechanical properties of natural fibre composites have been studied
previously by many authors. Oksman et al.11 investigated how poly(lactic acid)
acts as a matrix material for natural fibre composites using flax fibre, and
reported that the mechanical properties of poly(lactic acid) and flax fibre
composites are promising. The composite strength was about 50% better
compared to similar polypropylene/flax fibre composites, which are used today
in many industrial applications. The stiffness of poly(lactic acid) was increased
from 3.4 to 8.4 GPa with an addition of 30% flax fibres. Venkateshwaran
et al.24 studied the addition of sisal fibre to improve the mechanical properties
of the banana fibre/epoxy composite and reported that the addition of sisal up
to 50% increases the tensile strength, flexural strength and impact strength
around 16, 4 and 35%, respectively.
The plant fibre structure can be thought of as a composite material with the
stiff and strong cellulose microfibrils embedded in a hemicellulose/lignin
matrix. However, the composite structure in plant fibres is rather complex (e.g.,
two-phase matrix and cell wall layers). Moreover, plant fibres are part of a
larger biological system, i.e. the plants, with a long evolutionary history, and
their properties have therefore been highly optimized with respect to the
functional requirements of plants. Thus, the study of plant fibre mechanical
properties is not just an assessment of the reinforcement potential of plant
fibres in man-made composites, but might as well provide insight into the form
and function of a sophisticated composite material.
A number of structural aspects serve to restrain the practical attainable
tensile properties of plant fibres, e.g. the degree of cellulose crystallinity, the
microfibril angle and the cellulose content. Table 4.1 presents typical reported
tensile properties and microfibril angles of different types of plant fibres.
Stiffness and ultimate stress of hemp fibres have been reported in the ranges
30–60 GPa and 300–800 MPa, respectively. It can be seen in Table 4.1 that in
particular the measured ultimate stress of plant fibres is much below the
theoretical estimates. This might be explained by the presence of fibre defects,
which has been shown to affect the failure mechanisms in plant fibres.60 The
measured fibre stiffness more closely reflects the theoretical estimates. The
observed large variation in the measured tensile properties is a typical trait for
materials with a natural origin, but some variation is also added by the
experimental testing procedure. The measurement of tensile properties of single
Table 4.1 Microfibril angle and tensile properties of different plant fibres.a
Plant fibre Microfibril angle (1) Stiffness (GPa) Ultimate stress (MPa)
Hemp 6 30–60 300–800
Flax 6–10 50–70 500–900
Jute 8 20–55 200–500
Sisal 10–25 9–22 100–800
Softwood 3–50 10–50 100–170
Data for microfibril angles are from Gassan et al.,61 Anagnost et al.,62 Lilholt and Lawther63 and
a
Madsen.64
plant fibres is not a simple task, and problems are especially related to fibre
gripping and the determination of the fibre cross-sectional area.
4.4.6 Effect of Anisotropy in Natural Fibres on Mechanical

Properties
The mechanical properties of an anisotropic material are a function of
orientation. An anisotropic material has properties that vary with direction
within the material. This directional dependence is observed for other material
properties such as ultimate strength, Poisson’s ratio and thermal expansion
coefficient. Material property data, including fibre concentration, matrix
properties and properties of the composite in the principal material directions,
are incorporated into micro-mechanical models to estimate the fibre properties.
The determination of composite properties along the principal material
directions is, however, somewhat more involved. The mechanics concerning
coordinate system transformations can be applied to arrive at the well-known
relationship between off-axis Young’s modulus and the principal properties of
a 2-D composite ply.57 Young’s modulus describes tensile elasticity or the
tendency of an object to deform along an axis when opposing forces are applied
along that axis; it is defined as the ratio of tensile stress to tensile strain. It is
often referred to simply as the elastic modulus.
A broad spectrum of mechanical properties is provided by the selection of
reinforcement types, style, form and proportion in combination with a different
matrix. The directionality of strength in a composite can be greatly influenced
by substituting longitudinal reinforcement by random mats or directional
fabrics. The absolute value of the specific property desired depends on the fibre
type chosen: glass, carbon, aramid, organic or natural fibres. The mechanical
properties of composite plates reinforced by curvilinear fibres strongly depend
on the fibre shapes, and these composites exhibit non-uniform stiffness and
anisotropy. A very important feature of natural fibre composites is that
the mechanical properties of both the fibre and polymer matrix are time-
dependent.56
Neagu et al.65 evaluated a quantitative analytical–experimental method to
determine the stiffness properties of wood fibre from macroscopic tensile tests
of composites. This method can be divided into three parts: (1) measurement of
the Young’s moduli, fibre volume fraction and fibre orientation distribution in
the composites; (2) modelling of the Young’s moduli in terms of the anisotropic
elastic constants of the fibres using a laminate analogy and a micromechanical
model; and (3) inverse modelling to estimate the elastic properties of the fibres
from composite stiffness. The analytical–experimental method was employed to
estimate the longitudinal Young’s modulus of the fibres to evaluate their
potential reinforcement in composites. A correlation between lignin content
and fibre longitudinal Young’s modulus was observed, and an optimal lignin
content range at which fibre stiffness attains a maximum was identified for
softwood kraft fibres.
78 Chapter 4
55
Michalowski and Cermák reported that the distribution of fibre orientation
in practical applications is clearly anisotropic and that the use of existing iso-
tropic models leads to inaccurate predictions of the strength gain attributed to
fibres. For cases in which the predominant load is perpendicular to the preferred
plane of fibre orientation, isotropic models will, in general, underestimate
the utility of the fibres. These authors reported an increase in the strength of
fibre-reinforced soils due to anisotropic hardening. They reported that when
fibre-reinforced soil is subjected to large strains, the variation of fibre orientation
distribution produces an anisotropic increase in strength. According to Lakes,49
anisotropic composites offer superior strength and stiffness in comparison with
isotropic ones. Material properties along a certain direction are enhanced at the
expense of properties along other directions. It is sensible, therefore, to use
anisotropic composite materials only if the direction of the applied stress is
known in advance.
Ku et al.66 reported that the effect of hemp fibre content and anisotropy can
be examined on the basis of the tensile properties of the resultant composite
materials. The tensile strength, with fibres along the perpendicular direction,
tended to decrease with increasing hemp fibre content (a maximum decrease of
34% at 70% of hemp). The tensile strength of composites with fibres oriented
along the parallel direction showed a different trend, and a maximum value was
found with increasing fibre loading. These authors also reported that the tensile
strength of composites with fibres oriented along the perpendicular direction
was 20–40% lower than that of composites with fibres oriented along the
parallel direction. Because the fibres lay perpendicular to the direction of load,
they could not act as load-bearing elements in the composite matrix structure
and became potential defects that could cause failure. They described that the
best tensile properties were found in specimens cut from the composite sheets
parallel to the direction of carding.
John et al.12 studied the influence of fibre loading and fibre ratio on
mechanical properties. They reported that the longitudinal orientation of
fibres results in maximum tensile strength, and as the angle of orientation
of fibres increases, the tensile strength decreases. When fibres are longitudinally
oriented, the fibres are aligned along the direction of force and the fibres
transfer stress uniformly. When transversely oriented, the fibres are aligned
perpendicular to the direction of load and cannot take part in stress transfer.
The inclination angle relative to the fibre axis for cellulose microfibrils is a
key parameter to assess fibre tensile properties. The microfibril angle can be
determined by different techniques such as X-ray diffraction67 and by fungal
decomposition of the non-cellulose constituents in the cell wall.68 Madsen64
reported that both the strain and stress at which a composite fractures decrease
when the fibre axis is inclined to increasing angles with respect to the test direction.
This author evaluated the composite tensile strength and reported that there was a
four-fold decrease when the inclination angle to the fibre orientation was increased
to 261, whereas the composite stiffness only decreased by a factor of two.
Tensile properties of plant fibre composites compared to glass fibre com-
posites are presented in Table 4.2. Madsen64 reported that the composites with
Table 4.2 Tensile properties of plant fibre composites. For means of com-
parison, reported properties of glass fibre composites are included.
Fibre type Fibre orientation Stiffness (GPa) Ultimate stress (MPa) Ref.
Hemp Random 2.4–2.7 33–37 70
Aligned 27.6 277 64
Flax Random 3.4 36–39 71
Aligned 28.7 288 64
Jute Random 5.2 40–61 72
Aligned 27.2 225 73
Wood Random 4.2 28 74
Aligned 4.2 52 74
Glass Random 5.4 77 75
Aligned 45.0 1020 76
a random fibre orientation possess moderate tensile properties with stiffness

below 6 GPa and ultimate stress below 60 MPa. Table 4.2 shows that if the
fibres are aligned, the tensile properties are considerably improved. Thus,
(partial) alignment of plant fibres would be an alternative and cleaner approach
to improve composite properties, instead of treatment of fibres with chemical
additives. According to Table 4.2, the glass fibre composites are superior to
plant fibre composites irrespective of fibre orientation, and moreover, that
ultimate stress in particular is larger for glass fibre composites. Thus, Table 4.2
reveals the current status of plant fibre composites where stiffness is acceptable,
but ultimate stress needs to be somewhat improved. The lower weight of plant
fibre composites is usually used as an argument to compensate for their lower
properties. The lower density of plant fibres in relation to glass fibres (about 1.5
vs. 2.6 g cm3) is advantageous with respect to a lower composite component
weight. If tensile stiffness is selected as the key mechanical parameter, and
component stiffness is predetermined, it is anticipated that a composite com-
ponent based on plant fibres would possess the lowest weight. However, this
implies that the stiffness of plant fibres is similar to the stiffness of glass fibres,
which in general is not the case. The stiffness of glass fibres is reported to be
about 75 GPa.69 In the study reported by Madsen64 the estimated largest value
of plant fibre stiffness was about 65 GPa. Because of the difference in fibre
stiffness, the anticipations of composite component weight are not that
obvious. The difference in stiffness between plant and glass fibres is partly
equalized in composites with a random fibre orientation.
The structural anisotropy of natural fibres can be estimated through a cor-
relation with the mechanical properties of composites. The relationship between
the molecular and supramolecular structures of regenerated cellulose fibres and
their mechanical properties in the direction of the fibre axis has been studied
by Eichhorn et al.60 In the case of well-oriented cellulose fibres, the fibre com-
pliance is related to the orientation parameter hsin2 fiE by eqn (4.1):
1 1 hsin2 fiE
¼ þ ð4:1Þ
E ec 2g
80 Chapter 4
Where E is the modulus of elasticity in fibre direction, ec is the chain modulus
and g is the average shear modulus between adjacent chains.77 The strain
orientation parameter hsin2 fiE represents the average orientation of all
polymer chains at zero strain. This orientation parameter is equivalent to the
orientation parameter hsin2 fi (eqn 4.2), which is the second moment of the
molecular orientation distribution, only in well-oriented fibres with a Gaussian
distribution of chain orientation:78

2 2 Dn
hsin fi ¼ 1 ð4:2Þ
3 Dnmax
In eqn (4.2), Dn is the measured birefringence and Dnmax is the maximum

birefringence of cellulose. The validity of the model shown in eqn (4.1) for well-
oriented regenerated cellulose fibres was confirmed by Kong and Eichhorn.79
However, in order to fully understand the behaviour of cellulose fibre rein-
forcement in polymer composites, it is also necessary to study the mechanical
fibre properties transverse to the fibre direction. Gindl et al.77 studied the
changing ratio of the modulus of elasticity of regenerated cellulose fibres par-
allel and transverse to the fibre direction, independent of their degree of pre-
ferred molecular orientation. By fitting an empirical mathematical relationship
to the experimental data, the modulus of elasticity transverse to the direction of
the cellulose chain was inferred, and an estimate of the anisotropy (i.e., the ratio
of the modulus of elasticity parallel and transverse to the fibre direction) of a
regenerated cellulose fibre was made from the degree of preferred orientation of
its constituent cellulose chains. In this study an empirical model was developed,
which allowed making an estimate of the anisotropy of regenerated cellulose
fibres based on their degree of preferred orientation expressed by birefringence.
A mathematical function of the form was used and the parameters a, b and c
were determined by least-squares fitting (eqn 4.3). The calculated anisotropy
of regenerated cellulose fibres characterized in this study ranged from 3.1
for textile viscose fibre up to 15.3 for the experimental Bocell fibre.
E ¼ a þ b þ eðcDnÞ ð4:3Þ
4.5 Applications of Natural Fibre Composites

Natural fibre-containing composites are more environmentally friendly and
are used in transportation (automobiles, railway coaches, aerospace), military
applications, building and construction industries (ceiling panelling, partition
boards), packaging, consumer products, handicraft articles, fancy goods, etc.
Natural fibre composites are also used in the building and construction
industries as panels for partitions and false ceilings, partition boards, wall,
floor, window and door frames, roof tiles, and mobile or pre-fabricated
buildings, which can be used in times of natural calamities such as floods,
cyclones and earthquakes. Regarding storage devices, composites can be
used in post-boxes, grain storage silos and bio-gas containers. Fibre composites
can also be applied in furniture, electric devices, lampshades, suitcases, helmets
and toys.
According to Tudu,18 natural fibres are used in the automotive industry
because of their low density, which may lead to a weight reduction of 10–30%,
acceptable mechanical properties, good acoustic properties, favourable pro-
cessing properties, such as low wear on tools, possibilities for new production
technologies and materials, favourable accident performance, high stability and
less splintering, favourable ecobalance during vehicle operation due to weight
savings, occupational health benefits compared to glass fibres during produc-
tion, lack of off-gassing of toxic compounds (in contrast to phenol resin bonded
wood and recycled cotton fibre parts), reduced fogging behaviour and cost
effectiveness, both for the fibres and the applied technologies.
Natural fibres have replaced synthetic fibres as reinforcement in various
matrices. Natural fibre composites can effectively be used as substitutes for
wood and in various other technical applications, e.g. automotive parts. Tex-
tiles, ropes, canvas and also paper can be made of local natural fibres, such as
flax and hemp. The number of applications of natural fibre composites has
multiplied in recent years owing to their biodegradability and high specific
properties. The use of natural fibres in composite applications is being pursued
extensively throughout the world. Consequently, natural fibre composite
materials are being used to produce many components in the automotive sec-
tor. These materials are based largely on polypropylene or polyester matrices,
which incorporate fibres such as flax, hemp or jute.43
Owing to their light weight, high strength-to-weight ratio, corrosion resis-
tance and other attractive properties, natural fibre-based composites are
becoming important in building and civil engineering fields as panels for par-
titions and false ceilings, partition boards, walls, floors, window and door
frames, roof tiles and in mobile or pre-fabricated buildings. Despite their
usefulness in practice, synthetic fibre-based composites are difficult to recycle
after their designated service life. Moreover, natural fibre-based composites are
environmentally friendly to a large extent. Bamboo is a very well-known and
popular construction material, particularly in bamboo-rich regions. Sisal fibres
obtained from the leaves of sisal plants have also proven to be very suitable
reinforcement material in various polymeric matrices.80 Composites made from
soy oil-based resin and cellulose fibres have been used in roof structures.
Recycled paper was previously tested in composite sheets and structural unit
beams and was found to produce the required stiffness and strength required
for roof construction.81
4.6 Final Considerations

Based on various economic and environmental considerations, the commer-
cialization of composites reinforced with natural fibres should continue to
increase. New environmental regulations and societal concern have triggered
the search for new products and processes that are environmentally friendly.
The incorporation of natural fibres into composite materials can reduce the
82 Chapter 4
dependence on non-renewable materials. On the basis of economic and envir-
onmental considerations, the use of natural fibres will continue to increase in
markets owing to advantages of its characteristics of biodegrability. Studies on
specific properties of fibres, such as composition, molecular structure, source,
orientation, length, anisotropy and their effect on mechanical properties, are
important for applications in composites. Mechanical properties such as tensile
strength, Young’s modulus and elongation at break of natural fibre composites
are a function of orientation of the fibres in the matrix. Therefore, the struc-
tural anisotropy of fibres is related to the mechanical properties.
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CHAPTER 5
Flame Retardant Characteristics

of Natural Fibre Composites
BALJINDER K. KANDOLA
Institute for Materials Research and Innovation, University of Bolton,

Bolton, BL3 5AB, UK
5.1 Introduction
Composites are advanced engineering materials created by the synthetic
assembly of two (a matrix and a reinforcing element) or more (other additives)
components in order to obtain specific properties. The reinforcing component
provides mechanical strength and stiffness to the composite, whereas the matrix
helps in holding the reinforcement in the required shape and form. Since the
mechanical properties are governed by the reinforcing component, its choice is
crucial and depends upon the end-product requirements. The composite con-
struction technology is not a recent invention; on the contrary, it has evolved
over centuries following inspiration from materials existing in nature, such as
wood, plants, mammalian bones, etc. Wood, for example, is a composite
consisting of high-strength cellulosic fibres and a matrix of lignin. In terms of
composites known today, the use of fibres/fabrics as a reinforcing phase started
about 100 years ago,1 mostly as wood products with simple and relatively cheap
components. Prior to and during World War II, wood products and composites
were commonly used in aeroplanes and automotives. In 1941 a car was built
with wood, hemp, sisal and wheat straw, driven by hemp oil.2 All this changed,
however, with the development of high-performance fibres such as glass,

86
Flame Retardant Characteristics of Natural Fibre Composites 87
carbon and aramid. Owing to their excellent mechanical and low-density
properties, these high-performance fibres and derived composites became very
popular in aerospace, marine, automotive and construction industries. Over the
past decade or so there has been a renewed interest in natural fibre composites,
mainly due to a big push from policy makers of many European countries to
force the automotive industry to reuse and recycle materials. This has led to the
use of bio-based materials in automotive manufacturing.
At present their largest usage is in the automotive industry, mainly for semi-
structural applications. Semi-structural are those which require the component
to support its own weight in addition to bearing light external loads such as soft
knocks or impacts.3 In automotives, natural fibre composites are used for
seatbacks, headrests, front door-liners, rear door-liners, parcel shelves, boot-
liners, sunroof interior shields and noise, vibration and harshness materials.4
Flax, hemp, sisal, wool and other natural fibres are used in the Mercedes
Benz E-class vehicle. Daimler-Benz uses flax and abaca in dashboards, centre
armrests, seatbacks; BMW uses flax and sisal in interior door linings, kenaf and
flax in package trays and door panels. Similarly, Toyota and Ford use kenaf
fibres. In most cases the matrix is polypropylene (PP) or polyethylene (PE);
however, poly(lactic acid) (PLA) is also becoming popular.4 Although these
fibres with high-performance resins can form composites capable of passing
performance criteria for automotive exteriors, the high moisture absorption
behaviour of these composites has restricted their use as exterior components.
Mercedes, however, has started using abaca and flax fibre for exterior com-
ponents such as front bumpers. In the construction industry, natural fibre
composites (NFCs) are being used for door panels, window frames, office
cabinets, marine flooring, etc. For structural, load-bearing applications, nat-
ural fibre composites have had limited success,3 mainly due to their inferior
mechanical properties compared to high-performance fibres such as glass,
carbon and aramid.
Natural fibres from plant origins have certain advantages over glass fibres, in
particular low density, low abrasive wear, availability worldwide and renewable
and biodegradable nature. Their production is economical and they can be
easily recycled. Physical and mechanical properties of selected natural fibres are
given in Table 5.1 and compared with those of high-performance fibres com-
monly used for structural composites, such as glass, carbon and aramid.
Although the theoretical strength of natural fibres is very high (the theoretical
elastic modulus of the cellulose molecule5 is 138 GPa), in practice the strength
is very low (see Table 5.1), mainly due to many naturally occurring structural
defects within the fibre and those produced during fibre processing. Their
impact strength, in particular, is low compared with high-performance and
other synthetic fibres.6 Natural fibres, however, show higher elongation-at-
break than glass or carbon fibres, which may be good for the performance
of composites.7 Their thermal conductivity is low (0.29–0.32 W m1 K1) and
therefore they might make a good thermal barrier.7 There are, however,
some disadvantages of using natural fibres, such as low compatibility with
hydrophobic polymer matrices, thermal sensitivity at the temperature of
88 Chapter 5
6–8
Table 5.1 Physical and mechanical properties of fibres.
Tensile Initial Elongation Moisture
Diameter strength modulus at break Density absorption
Fibre (mm) (GPa) (GPa) (%) (kg m3) (%)
Cotton 16–21 0.3–0.6 5.5–12.6 2.0–10.0 1500–1600 8–25
Flax 40–620 0.3–1.5 27–80 1.2–3.2 1400–1500 7
Hemp 16–50 0.5–1.1 3–90 1.3–4.7 1400–1500 8
Jute 30–140 0.2–0.8 3–55 1.4–3.1 1300–1500 12
Ramie 40–60 0.4–0.9 44–128 2–3.8 1500 12–17
Coir 100–450 0.1–0.3 3–6 15–47 1250–1500 10
Sisal 100–300 0.5–0.9 9–28 2–3 1300–1500 11
Abaca 17–21 0.98 72 10–12 1500 –
E-Glass 3–20 2–6 50–100 2.5–3 2400–2600 –
S-Glass 3–20 3.5 87 2.5 2500 –
Carbon 5.0–6.9 1.5–7.0 150–240 1.4–1.8 1400–2000 –
Para- 10–12 2–4 63–150 3.3–3.7 1400–1450 5–7
aramid
compounding processes, moisture absorption and flammability. To improve

the adhesion between hydrophilic fibres and hydrophobic matrices, the fibres
are usually surface treated or compatibilizers are added in the polymer matri-
ces. Moisture absorption can also be reduced by surface treatments. Thermal
stability, flammability and smoke generation properties of composites pro-
duced from these fibres, however, are also major issues these days, because,
depending upon applications, they must pass some type of regulatory fire test
in order to ensure public safety. This is more important if their usage is to be
expanded in the marine and aerospace sectors, where fire regulations are more
stringent than in the automotive sector. In this chapter the key fire issues
pertaining to the thermal stability and flammability of composites and
their components, the fire retardant requirements for their usage in different
sections and flame retardant solutions are discussed. This chapter also com-
plements previous book chapters on flammability and fire retardancy of
high performance fibre-reinforced polymer (thermoset) composites.8,9
5.2 Thermal Stability and Flammability of

Natural Fibres
Natural fibres from plant origins can be grouped into three categories: seed
hairs (e.g. cotton); bast or skin fibres, collected from the phloem (the inner bark
or the skin) or bast surrounding the stem of plants (ramie, jute, flax etc.); and
leaf fibres (sisal and abaca). Out of these, jute, ramie, flax and sisal are most
commonly used for composites.10 All of these fibres consist of highly crystalline
cellulose fibrils spirally wound in a matrix of amorphous hemicelluloses, lignin
and pectin. Lignin and pectin act as bonding agents.
Chemically, cellulose contains glucose units linked together to form long
unbranched chain with 1,4-b-glycosidic linkages. The thermal degradation of
cellulose has been extensively studied in the literature11 and is summarized here
in Scheme 5.1. It proceeds through two types of reactions. At lower tempera-
tures (200–280 1C), there is gradual degradation which includes depoly-
merization, dehydration and decarboxylation leading to formation of
‘‘dehydrocellulose’’. This ‘‘dehydrocellulose’’ decomposes further to various
volatile products and char. At higher temperatures (280–340 1C), a rapid
volatilization occurs and a non-volatile liquid intermediate product, laevoglu-
cosan, is formed. Laevoglucosan subsequently pyrolyses to lower molecular
weight and highly flammable volatiles, and leaves a charred residue. Hence slow
heating favours the first type of reactions, leaving more char, whereas fast
heating produces more tar and less char. This is also the basis of imparting
flame retardancy to cellulosics, where condensed-phase active flame retardants
promote the first type of reactions, producing more char at the expense of
volatile combustible products.
Hemicellulose contains short, highly branched chains of sugars. It contains
five-carbon sugars (D-xylose and L-arabinose), six-carbon sugars (D-galactose,
6
D-glucose and D-mannose) and uronic acid. Whilst cellulose is crystalline,
strong and resistant to hydrolysis, hemicellulose has a random, amorphous
structure with little strength. It decomposes between 200 and 260 1C but forms
more non-combustible gases and less tar than cellulose,12 mainly going through
the first type of reactions shown in Scheme 5.1.
Lignin is formed by the removal of water from sugars to form aromatic
structures. There are many monomers of lignin; the type depends on the
source in nature. Large lignin molecules are three-dimensional and highly
cross-linked.6 Lignin starts decomposing from about 160 1C and continues to
decompose until about 400 1C. At the lower temperatures, relatively weak
200–280 °C
Dehydrocellulose Char + Gaseous products
Slightly Exothermic
endothermic
Cellulose
Exothermic Decomposition Flammable volatiles
280–340 °C Tar
(Mainly
Exothermic Laevoglocosan)
Char
Exothermic Dehydration
Scheme 5.1 Various stages of the pyrolysis of cellulose.

90 Chapter 5
bonds break, whereas at higher temperatures, cleavage of bonds in the aromatic
rings of the lignin takes place.12 Lignin contributes more to char formation
than either cellulose or hemicellulose.12
From the above discussion it is clear that fibres having higher cellulose
content should have higher flammability than the ones with higher lignin
content. The compositions of different fibres are presented in Table 5.2. Hence,
coir, kenaf, sisal and jute should be less flammable than other fibres. However,
their onset of decomposition temperature determines their suitability for usage
with a particular polymer matrix of the thermoplastic composite.
The thermal stability of natural fibres can be studied by thermogravimetric
analysis (TGA). Kozlowski et al.13 have reported the TGA of two bast fibres,
hemp and flax (lignin content 2–5%), and compared them with two leaf fibres,
cabuya and abacca (lignin content 7–13%). All fibres started degrading at
about 230 1C with mass loss curves very similar, except that the decomposition
process of bast fibres was a one-step process, whereas for the two leaf fibres
it was a two-step process, where the lower temperature (230–280 1C) step was
attributed to hemicellulose decomposition and the higher one (280–300 1C) to
lignin decomposition. They also studied their flammability by cone calorimetry
at 35 kW m2 heat flux. Hemp and flax fibres showed a lower peak heat release
rate and mass loss rate compared with abacca and cabuya, which is contrary to
the above discussion for the stability of the lignin content. They have argued
that during the decomposition of lignin, relatively weak bonds break at lower
temperatures, whereas cleavage of bonds in the aromatic rings takes place at
higher temperatures. Hence, with a lower lignin content the degradation begins
at a higher temperature, but there is a lower oxidation resistance, which would
be provided by the aromatic structure of lignin. Similar observations were made
by Manfredi et al.14 for flax, sisal and jute fibres, with a lignin content of 2.0, 9.9
and 11.8%, respectively. These fibres in composite form using an acrylic resin
showed different behaviour against fire in a cone calorimeter test. The com-
posites containing flax and sisal caused a long duration but slow growing fire.
On the other hand, jute fibre composites caused a quickly growing but short
duration fire.
Table 5.2 Chemical composition of fibres.6

Fibre Cellulose (%) Hemicellulose (%) Lignin (%) Pectin (%)
Flax 60–81 14–19 2–3 1.8–2.3
Jute 51–72 12–20 5–13 0.2
Abaca 61–64 21 12 0.8
Sisal 43–88 10–13 4–12 0.8–2
Kenaf 36 21 18 2
Ramie 69–76 13–15 0.6–1 1.9–2
Hemp 70–78 18–22 4–5 0.9
Cotton 83–92 2–6 0.5–1 5.7
Coir 43 0.3 45 4
Wood 45–50 23 27 –
(a) (b)
Figure 5.1 (a) TGA and (b) DTA curves of sisal, jute and wool fibres in air.
From our own work the TGA and DTA curves for jute and sisal fibres
are shown here in Figure 5.1 and compared with those of wool fibres. The onset
of the decomposition temperature of jute (235 1C) is much higher than sisal
(209 1C) and wool (204 1C) fibres. Wool fibre, however, starts decomposing
earlier and is more thermally stable in terms of mass loss than both cellulosic
fibres. The DTA curves also show that exothermic peaks representing
decomposition and char oxidation reactions for both cellulosic fibres are at
lower temperatures and are of higher intensity compared to wool fibres, sug-
gesting that they should be more flammable than wool fibres.
The crystallinity and orientation of fibres also affect their flammability. For
cellulosic fibres, higher levels of crystallinity result in higher levels of lae-
voglucosan during pyrolysis and, hence, increased flammability.15 Increasing
orientation results in decreased pyrolysis and therefore decreased flamm-
ability.16 However, most of these studies reported in the literature are in terms
of TGA measurements. How much these factors affect the flammability of
fibres in terms of the limiting oxygen index, or vertical flame spread, is not
known.
To compare the flammability of fibres, the limiting oxygen index (LOI) test
is good indicator. LOI is the minimum concentration of oxygen, expressed as
volume percent, in a mixture of oxygen and nitrogen that will just support the
flaming combustion of a material.17 Usually the fibres are tested as textile
structures. There is no standard method of determining the LOI of fibres. In
our laboratory we have developed our own methodology for testing fibres,
where the fibre is rolled into slivers about 10–12 cm long. The LOI test is
undertaken in a manner similar to that used for plastic sticks. The LOI value
of cotton is 18–20 vol%, and while the LOI values for other fibres are not
available in the literature, these are expected to be in a similar range, with the
higher lignin content being higher than cotton by 1–3 units. Hence, all these
fibres are flammable, more so than protein fibres such as wool with a 25 vol%
LOI value. The other flammability test for textiles is the vertical flame spread,
e.g. BS 5438:1989,18 which is applicable for textile structures. There is no study
available in the literature where the vertical flame spread of fabrics from
different natural fibres of similar area densities and weave/knits have been
compared.
92 Chapter 5
5.3 Flammability of Composite Matrices
Natural fibres can be used with thermoset, thermoplastic or biopolymers as
long as the processing temperature of the polymer is lower than the onset of
the decomposition temperature of the lignocellulosic fibre used. Natural fibres
start decomposing around 200 1C, as discussed in the above section. Their use
with different types of matrices reported in the literature has been summarized
in Table 5.3.10,12,19–30
5.3.1 Thermoplastics
While the physical, thermal and flammability properties of polymers in general
have been extensively reviewed in the literature, selected properties taken
from different sources31–36 are given in Table 5.4. The softening temperatures
and melting temperatures of thermoplastics are used for their processing, and
hence determine the suitability of their use with natural fibres. The only
thermoplastic polymers processable below the decomposition temperature of
Table 5.3 Reported natural fibre–matrix combinations to produce

composites.a
Biodegradable Thermoset
Natural fibresb Thermoplastic matricesc matricesd matricese
Wood flour/ PP, PE, PVC, – UP
fibre PS, PU
Cellulose PP, PE, PA66, PLA Ep
PVC, PS
Jute PP, PE PLA SBR, rubber, Ep,
UP, VE, Ph,
acrylic
Flax PP, PE PLA, PGA, PCL, Ep, VE, Ph, mela-
PHB mine, acrylic
Sisal PP, PE, PS – Rubber, Ep, UP
Abaca PP PLA, PHBV Ep
Kenaf PP, PE PLA Ep, Ph
Ramie PP PLA –
Hemp – – Ep, UP, VE, Ph
Coir – – Rubber, UP, Ep
Banana – – UP, Ph
Bamboo PP – Ep, UP
Pineapple PE – UP, Ph
Sunhemp PP – UP
Wheatstraw PP – –
Wool – – UP
a
PP ¼ polypropylene; PE ¼ polyethylene; PS ¼ polystyrene; PVC ¼ poly(vinyl chloride);
PU ¼ polyurethane; PLA ¼ poly(lactic acid); PGA ¼ poly(glycolic acid); PCL ¼ (poly-e-capro-
lactone), PHB ¼ poly(hydroxybutyrate); PHBV ¼ poly(3-hydroxybutyrate-co-3-hydroxyvalerate);
Ep ¼ epoxy; UP ¼ unsaturated polyester; VE ¼ vinyl ester; Ph ¼ phenolic.
b
Onset of decomposition temperature of natural fibres is in the range 200–230 1C.
c
Taken from references 10, 12 and 19 and references cited therein.
d
Taken from references 19–28 and references cited therein.
e
Taken from references 10, 12, 19, 29 and 30 and references cited therein.
Flame Retardant Characteristics of Natural Fibre Composites
Table 5.4 Thermal and flammability characteristics of selected thermoplastics.31–36
Glass Cone calorimetric datae at 40 kW m2
transition Melting Decomp. Heat of
temp.a temp.b temp. combust.c LOId PHRR THR
Polymer (1C) (1C) rangec (1C) (MJ kg1) (vol%) TTI (s) (kW m2) (MJ m2)
Polyethylene (–125)–2 137–146 340–440 48.0 18 159 1408 221
Polypropylene (–35)–1 160–220 330–410 44.0 18 86 1509 207
Poly(methyl 94–150 160–220 170–300 26.0 18 36 665 828
methacrylate)
Poly(vinyl 69–98 177–212 200–300 18.0 45 73 175 24
chloride)
Polyamide 6 40–100 210–220 300–350 32.0 23 65 1313 226
Poly(butylene 31 220–267 – – 22 113 1313 170
terephthalate)
Poly(ethylene 60–86 265–310 285–305 21.5 25 116 534 114
terephthalate)
Polycarbonate 145–150 220–267 350–400 31.0 27 182 429 119
Polystyrene 100–109 177–277 300–400 40.0 18 97 1101 210
Poly(tetrafluoro- (–113) 327–346 510–540 4.6 95 10000 13 12
ethylene)
ABS 110–125 – – 35.0 19 69 944 163
SAN 100–200 – – 36.0 – – – –
a
Taken from references 34 and 35.
b
Taken from references 34–36.
c
Taken from reference 33.
d
e
93
94 Chapter 5
natural fibres are polyethylene (PE, mp 137–146 1C), polypropylene (PP, mp
160–220 1C), polystyrene (PS, mp 177–277 1C) and poly(vinyl chloride) (PVC,
mp 177–212 1C). As seen from Table 5.3, these are most commonly used
polymers. Other polymers such as polyamide 6 (PA6, mp 210–220 1C),
polyester (PET, mp 265–310 1C), polycarbonate (PC, mp 220–267 1C), etc., are
processable at temperatures where natural fibres start decomposing. Out of
these polymers, PP is a more popular choice, usually with short fibres, where
composites are produced by melt blending and subsequently compression or
injection moulding.
The LOI and cone calorimetry are common methods of quantifying the
flammability of polymers. LOI has been discussed in the previous section. In a
cone calorimeter, samples in a horizontal orientation are exposed to an external
heat flux (25–100 kW m2) and ignited with a spark ignition, as detailed in
ISO-5660.37 Important parameters that describe the flammability behaviour of
polymers include the time to self-sustained ignition (TTI), the heat release rate
(HRR) and its peak value (PHRR), the total heat release (THR), the amount of
smoke generated and the residual char yield. The LOI31 and cone calorimetric
results for selected thermoplastic polymers tested by Hirschler32 at 40 kW m–2
heat flux are given in Table 5.4. The results show that PE, PP and PS, com-
monly used for natural fibre composites, are highly flammable, their LOI values
being 17–18 vol%, which are very low compared to some other polymers. The
peak heat release (1100–1510 kW m2) and total heat release (207–220 MJ m2)
of these three polymers studied at 40 kW m2 heat flux32 are very high com-
pared to others. However, PVC, which also can be used with natural fibres,
is inherently flame retardant (LOI ¼ 45 vol%), with very low PHRR and THR
(see Table 5.4). PVC contains chlorine in the polymer backbone, which
provides inherent flame retardant characteristics. PVC, however, releases HCl
during burning, so has environmental issues associated with it.
5.3.2 Biodegradable Matrices

Biodegradable materials are those which are capable of decomposing into
carbon dioxide, methane, water, inorganic compounds or biomass by the
enzymatic action of microorganisms. Biodegradable matrices include PLA,
cellulose esters, starch plastic, poly(e-caprolactone) and aliphatic polyester-
co-polyesters.6 All of these biopolymers are hydrophilic polyesters and so they
absorb moisture. In the case of natural fibre reinforcement, this could be even
more problematic since cellulosic fibres tend to absorb rather large amounts of
water when exposed to it. These biopolymers are mostly used in the packaging
industry and other applications with minor strength requirements. PLA com-
posites, however, are finding applications in other sectors as well, mainly
automotive. Their use with different natural fibres in composites, as reported in
the literature,19–28 is shown in Table 5.3.
PLA is a linear aliphatic polyester derived from renewable sources such as
corn.38,39 PLA is produced from starch, extracted from plant matter (e.g. corn),
which is converted into fermentable sugar (e.g. glucose) by enzymatic
hydrolysis. This natural sugar then, through fermentation, is converted into
lactic acid, as shown schematically in eqn (5.1):
enzyme hydrolysis þ H2 O fermentation polycondensation or

Starch ! Glucose ! Lactic acid ! PLA
ring-opening polymerization
ð5:1Þ
PLA’s properties in fibre form have been reviewed by Farrington et al.39 Its
mechanical properties are similar to conventional PET fibre.39 Although the
fibre is flammable, its flammability is slightly less (LOI ¼ 26 vol%) compared to
PET fibre (LOI ¼ 22–25 vol%).39 In matrix form the physical and mechanical
properties of PLA have been reported by Van de Velde and Kiekens20 and
other researchers.21–28 PLA’s glass transition temperature is between 60 and
65 1C and melting temperature between 173 and 178 1C. Since the processing
temperature is lower than the decomposition temperature of most natural
fibres, this can be easily used for preparing composites. Its density, however, is
higher than PP.20
In our recent research work, we have studied the flammability of PLA as a
matrix polymer and compared it with that of PP. Nonwoven mats of PP and
PLA fibres were used for nonwoven web production. These were melt pressed
at 190 1C for 2.5 min, followed by quick cooling to obtain 2.5–2.7 mm thick
sheets. TGA and DTA of the composites were performed on an SDT 2960
simultaneous DTA–TGA instrument from room temperature to 700 1C using
15 1 mg samples heated at a constant rate of 10 1C min1 in air flowing at
100 5 mL min1. The results are shown in Figure 5.2.
The DTA profile of polypropylene (Figure 5.2b) shows a melting endotherm
peak at 169 1C, followed by a large exotherm with a peak maximum at 382 1C
representing the main decomposition step. This is followed by another very
small exotherm at 423 1C, which represents the oxidation of the products
formed in the first step. The TGA curve (Figure 5.2a) shows one large
decomposition stage in the temperature range 197–395 1C, with 98% mass loss.
Between 395 and 480 1C, 2% mass loss occurs, representing the oxidation of
combustible products. No char is left at the end of the test.
(a) (b)
Figure 5.2 (a) TGA and (b) DTA curves of PP and PLA matrices in flowing air.
96 Chapter 5
The DTA of PLA (Figure 5.2b) shows an endotherm at 58 1C, representing
the glass transition temperature, an endotherm at 169 1C, representing melting,
an endotherm at 342 1C followed by a double peaked exotherm (362, 376 1C),
representing the main decomposition, and an exotherm at 455 1C, representing
the oxidation of char. The TGA curve (Figure 5.2a) shows that the onset of
decomposition occurs at 296 1C and 97% mass loss occurs up to 385 1C,
followed by 3% up to 465 1C. On comparing the TGA and DTA curves of PP
and PLA in Figure 5.2, PLA seems to be more thermally stable than PP.
These samples were also tested by a cone calorimeter at 35 kW m2 heat flux.
Mass loss, heat release rate and smoke production versus time curves are shown
in Figure 5.3 and derived results are presented in Table 5.5. Both samples
ignited at between 28 and 30 s. PP showed a PHRR of 1699 kW m2 and a THR
of 95.4 MJ m2. For PLA the values are much lower: PHRR ¼ 663 kW m2 and
THR ¼ 49.8 MJ m2. The effective heat of combustion of PLA (18.2 MJ kg1)
is also much lower than for PP (42.9 MJ kg1). The trend for mass loss for PP
and PLA samples (Figure 5.3a) is different from that observed by TGA, as here
the mass loss curves are quite similar. This is due to the high heat flux, causing
rapid degradation. PLA produces negligible smoke compared to PP, as shown
in Figure 5.3(c). This study shows that although PLA is combustible, it poses
less of a fire hazard as it releases less heat during combustion.
5.3.3 Natural Rubber

Natural rubber is an elastomer, commonly derived from latex of many plants
and trees. Chemically it is polyisoprene, mainly cis-1,4-polyisoprene, although
(a) (b)
(c)
Figure 5.3 (a) Mass loss, (b) HRR and (c) smoke vs. time curves for PP and PLA
matrices exposed to 35 kW m2 heat flux.
Table 5.5 Cone calorimetric results for PP, PLA and different fibre-reinforced
composites at 35 kW m2 external heat flux.
Sample Hc
thickness TTI FO PHRR THR (MJ Smoke Char
Sample (mm) (s) (s) (kW m2) (MJ m2) kg1) (m2 m2) (%)
PP 2.7 30 160 1699 95.4 42.9 1302 0
PLA 2.5 28 180 663 49.8 18.2 0.7 3.0
Glass/PP 3.8 25 260 506 79.7 38.1 1308 50.9
Glass/PLA 3.6 27 280 275 46.4 16.6 0.1 43.9
Jute/PP 3.2 33 290 675 95.5 31.7 1121 3.3
Jute/PLA 2.8 38 300 393 60.7 16.1 0.9 4.4
Sisal/PP 1.3 25 140 680 43.1 30.6 532 3.2
Sisal/PLA 1.3 27 120 542 28.1 15.9 1.2 3.3
Wool/PP 3.6 30 360 632 140.9 35.1 1928 4.6
the trans isomer can also be isolated from some plants. Both of these forms
can also be produced synthetically. Natural rubber is very flammable, with
LOI ¼ 17 vol%.31 Rubber is generally vulcanized before use. The ingredients
for vulcanization may increase or decrease the flammability of rubber, but
generally increase the smoke production and toxicity.31
5.3.4 Thermosets
Thermoset matrices are fabricated from the respective resin, a curing agent, a
catalyst or curing initiator and a solvent sometimes introduced for lowering the
viscosity and improving the impregnation of reinforcements. In thermosets,
solidification from the liquid phase takes place by the action of an irreversible
chemical cross-linking reaction which produces a tightly bound three-
dimensional network of polymer chains. The molecular units forming the
network and the length and density of the cross-links of the structure will
influence the mechanical and thermoplastic or non-thermoplastic and flamm-
ability properties of the material. The level of cross-linking between resin
functional groups and often the degree of non-thermoplasticity is a function of
the degree of cure, which usually involves application of heat and pressure.
However, some resins cure at room temperature.
The most common types of thermoset matrices for composites are unsatu-
rated polyester, vinyl esters and phenolics. All of these matrices used with
different natural fibres and reported in literature are summarized in Table 5.3.
Unsaturated polyester resins are prepolymer mixtures containing unsaturated
groups and styrene, with the latter serving as both a diluent and cross-linking
agent during the radical polymerization process. Vinyl ester resins are similar in
structure to unsaturated polyester resins, differing only in the location of the
primary vinyl reactive groups, which are positioned at the ends of the mono-
meric chains. Both unsaturated polyester and vinyl esters are room temperature
curing resins, often post-cured at 80 1C, and hence are suitable for use with
98 Chapter 5
natural fibres. These, however, are very flammable with LOI values of
20–23 vol%,8,9,40 and burn with heavy smoke and soot.
Epoxies refer to a wide variety of cross-linked polymer chains based on
polymer monomeric units containing an epoxide group. Epoxy resins are
typically formed from the reaction between a diepoxide with a primary dia-
mine. Reactants with higher reactive functionalities result in a highly cross-
linked, stiff and tough epoxy network suitable for aerospace applications.9
These, however, are cured at higher temperatures than those with lower
functionalities. The latter are room temperature curing resins. The flammability
also differs: the ones with high functionalities are highly cross-linked and less
flammable than the low functionality ones. In our recent work we have tested
two resins with different functionalities;41 their flammability behaviour is
shown in Table 5.6. Epoxy resins generally have superior functionalities, which
result in better mechanical properties compared to unsaturated polyester resins
or vinyl esters, but their use is limited due to high cost which can be several
times higher than the latter.
Phenolics are inherently flame retardant resins, but they are brittle and
do not possess high mechanical properties. We have discussed the flammability
of these resins in detail in previous reviews,8,9,40 where it was shown that the
flammability of these resin can be ranked as:
Phenolic o polyimide o bismaleimide o epoxy o polyester and vinyl ester
It must be emphasized that this is a generic discussion on these resins. Since
thermosets contain different components (resin, curing agent and sometimes
hardener), the choice of components for each resin type will affect the
flammability properties of the cured resin, as shown for the two different epoxy
resins in Table 5.6. Hence, the trend is very generic. In most cases the flamm-
ability of low-temperature curing epoxies is similar to unsaturated polyester or
vinyl ester resins. Similarly, some phenolics are better than others.
Table 5.6 Flammability characteristics of selected thermoset matrices.41

Cone calorimetric data at
50 kW m2
Curing LOI TTI PHRR THR
Resin Formulation conditions (vol%) (s) (kW m2) (MJ m2)
Epoxy-1 Tetraglycidyl-4,4 0 - 180 1C for 3 h 27.8 24 823 56
diaminodiphenyl-
methane, cured
with 4,4 0 -
diaminodiphenyl
sulfone
Epoxy-2 Butane-1,4-diol 22 1C for 24 h, 21.5 34 1861 77
diglycidyl ether, post-cured
cured with at 80 1C
cycloaliphatic for 1 h
amines
5.4 Flammability of Composites
5.4.1 Performance Requirements Depending
Upon Application Areas
For the usage of natural fibre composites in the transport and construction
industries, it is essential to assess the fire risk associated with the application.
All products have to conform to certain specified regulations for particular
applications. A comprehensive list of international standards and codes used
for fire-safety design of fibre-reinforced polymer composites for different sec-
tors is beyond the scope of this chapter. The reader is referred elsewhere to such
reviews and book chapters;33,42,43 only selected relevant ones are discussed
here briefly.
For the automotive sector, in the USA the Federal Motor Vehicle Safety
Standard 302 (FMVSS 302)44 regulates the flammability of materials used in
the interiors of cars and trucks. The test is run on samples mounted horizon-
tally and ignited with a 3-mm diameter, 20-mm long Bunsen burner flame.
To pass this test the horizontal burn rate of the sample should not exceed
102 mm min1. The FMVSS 302 test is virtually an international standard, as it
has been harmonized with many equivalent designations.42 Other examples
include ISO 3795, BS AU 169 (UK), ST 18-502 (France), DIN 75200
(Germany), JIS D 1201 (Japan) and SAE J369 (automotive industry). This test
is not as stringent as in other sectors.
For the rail industry the rate of fire spread and toxicity of the fumes pro-
duced during their combustion are of vital importance. In the US the Federal
Railroad Administration (FRA) has a standard, Federal Register 49:192
(1984), which gives requirements for all materials used in rail carriages. In
Europe there is not a common standard across the different countries. Each
European country has its own regulations, e.g. DIN 5510 (Germany), BS6853
(UK) and NF F16-101 (France). Most of these include flame spread, smoke
development and toxicity. Other tests include the well-established BS 5852:2006
for upholstered furniture composites and/or complete pieces of furniture,
such as seats in rail compartments when they are subjected to a smouldering
cigarette, or to a flaming ignition source of thermal output ranging from a
burning match to that approximating to the burning of four double sheets of
full-size newspaper.45 On the other hand, smoke production may be determined
using ISO 5659-2:2006 based on the well-known NBS smoke chamber
method46 and is similar to ASTM E662.
For marine applications, all commercial passenger and cargo ships in Eur-
opean countries have to comply with the fire performance requirements con-
tained in the International Convention for the Safety of Life at Sea (SOLAS)47
as IMO/HSC Code (Code of Safety for High Speed Craft of the International
Maritime Organization). The fire tests to be carried out and the acceptance
criteria are defined in the IMO/FTP code (International Code for Application
of Fire Test Procedures), which have been mandatory since 1998.48 This
code allows for use of non-conventional shipbuilding materials, defined as
‘‘fire restricting’’, materials which have low flame spread characteristics, limited
100 Chapter 5
rate of heat release and smoke emissions. Furthermore, for areas of moderate
and major fire hazard (e.g., machinery spaces, storerooms), the materials used
should be ‘‘fire resisting’’, i.e. they should prevent the fire and smoke propa-
gation to adjacent compartments during a defined period of time (60 min for
high hazard and 30 min for low hazard areas). Composites used for load-
bearing structures should be able to maintain their load-bearing capacity within
the specified period of time (30 or 60 min). For fire-restricting material char-
acterization, specified tests include the room corner test, ISO 9705, and the cone
calorimeter test, ISO-5660.
For aerospace applications the polymer-based composites used for the
interior design of aircraft must be self-extinguishing and exhibit low flame,
smoke and toxicity (FST) characteristics; they are required to comply with
industry and international regulations such as FAR (the US Federal Aviation
Regulations), which govern the requirements for materials used for such
applications.49 The most important test is the Heat Release Rate Test (FAR
25.853, Part IV, Appendix F), according to which when the sample is tested at
35 kW m2 external heat in a Ohio State University calorimeter, both the peak
heat release rate and total heat release rates measured over 2 min should not
exceed 65 kW m2. For detailed descriptions of all other fire tests, the reader is
referred to the FAR regulations.
5.4.2 Key Parameters Affecting the Flammability of Composites

The physical, mechanical and flammability properties of composites not only
depend upon the matrix and fibre type, but also on the reinforcement type. The
latter also determines the manufacturing process. If short fibres of only a few
millimetres in length or particles such as cellulosic wood flour are used as the
reinforcing element, then for thermoplastic composites, melt mixing in a con-
tinuous extrusion process is often used. Both polymer and fibres are introduced
into the extruder as a means of optimizing the wetting and mixing needed to
obtain good fibre dispersion. With this technique the fibre content achieved is
usually less than 40% by weight of fibres owing to processing issues. The fibre
length is reduced further under the processing conditions, resulting in fibres
being oriented in complex and often non-optimal patterns.50 Since short
fibres function mainly as fillers, the flammability of the composite produced
should be different from the one where reinforcement is in fabric form. For
composites with woven fabric reinforcement, the polymer as a cast sheet or
extruded film is introduced between layers of reinforcement fabrics. The
assembly is passed through a series of rollers that melt the polymer and enables
impregnation. After a chilling roll, the impregnated material is collected as a
sheet. Commingling of reinforcing fibre and matrix in a fibrous form is another
common method of composite fabrication. On heating under pressure, the
matrix fibres melt and a consolidated structure is obtained.
For thermoset composites, short fibres are mixed with resin before casting
into moulds of the desired shape. In textile form, woven/knitted fabrics may be
impregnated with resins using techniques such as hand lay-up, resin transfer
moulding, etc., and then stacked to produce samples of desired thicknesses.
Although the effect of different reinforcement types on the flammability of the
composites is not documented in the literature, it is expected that the flamm-
ability of the resulting composites would be different.
Various other factors affecting the flammability of composites in general have
been reviewed previously.8,9,40 The composite thickness of a structure has a
significant effect on its flammability. Thermally and physically thin samples
ignite early, show large peak heat release values, burn for a shorter time and
produce less total heat release compared to thermally and physically thick
samples which burn slowly but for a longer time.51,52 ‘‘Thermally thick’’ means
the heat wave penetration depth is less than the physical depth, whereas in
‘‘thermally thin’’ the composite has the same temperature through this limiting
thickness. The transition from thermally thin to thermally thick is not a constant
since it depends on the material thermal properties, including fibre and resin
thermal conductivities. For a given composite of defined thickness, the condition
depends on the intensity of the fire or, more correctly, the incident heat flux.
While many large-scale fire tests involve heat sources or ‘‘simulated fires’’ having
constant and defined fluxes, in real fires, heat fluxes may vary. For example, a
domestic room filled with burning furniture at the point of flashover presents a
heat flux of about 50 kW m2 to the containing wall and door surfaces; larger
building fires present fluxes as high as 100 kW m2 and hydrocarbon fuel ‘‘pool
fires’’ may exceed 150 kW m2. The heat flux dependence on burning behaviour
can be examined using calorimetric techniques such as the cone calorimeter.8,9,40
The overall burning behaviour of a composite will be the sum of its com-
ponent fibres and thermoplastic polymer/resin plus any positive (synergistic) or
negative (antagonistic) interactive effects.
5.4.3 Flammability Data

Despite widespread interest in natural fibre composites and their increasing
usage in different sectors, there is little published literature on their fire per-
formance. This is mainly due to the fact that so far they have only been widely
used in the automotive industry and fire safety regulatory requirements for
automotives are not very stringent. All available literature is from the research
prospective, where mainly cone calorimetry has been used to assess their
flammability. Most of the work reported in the open literature and some
recently carried out in our laboratories is reviewed here in the following sections.
5.4.3.1 Thermoplastic Composites Reinforced with Short Fibres

A number of studies have been conducted on the burning behaviour of PP
composites reinforced with short fibres.53–56 In general, the presence of fibres
reduces the time-to-ignition, peak (PHRR) and total (THR) heat release rates,
but increases the total burning time or time-to-extinguishment in cone calori-
metric tests. PP shows one peak heat release rate, whereas in natural composites
two peaks are observed.53–56 The first major peak represents the surface igni-
tion and burning, and is usually reduced in intensity compared to pure PP
owing to the charring behaviour of lignocellulosic fibres. The second peak
102 Chapter 5
represents the char-surface rupture and eventual burn-out of the whole sample.
Borysiak et al.53 have reported the flammability behaviour of PP/wood com-
posite samples, prepared by co-extruding PP and Scotch pine wood particles
(50 wt%) in a single screw extruder and then press moulding them. The
flammability of the composites was tested in a cone calorimeter at 35 kW m2
external heat flux and compared with PP plaques of similar thicknesses. The
time-to-ignition (TTI) of the composite decreased from 47 s for pure PP to 31 s,
the PHRR from 1407 to 810 kW m2, the THR from 120 to 92 MJ m2 and the
average specific extinction area from 514 to 393 m2 kg1. This indicates that
natural fibres reduce the TTI of composites owing to their easily ignitable
characteristics. However, once ignited, they are not as flammable as hydro-
carbons, and hence reduce the overall flammability of the composites. Schartel
et al.36 and Le Bras et al.54 have studied flax/PP (40:60 wt%) composites
containing short fibres and reported that with fibres the PHRR decreased
to 640 kW m2 in composites compared to 1800 kW m2 in PP,54 when tested
at 50 kW m2 external heat flux.
Helwig et al.56 prepared PP/flax composites with long flax fibres
(150–300 mm), where fibres were placed between PP sheets and then melt
pressed. A number of composites with fibre contents of 12.5, 20, 30 and 40%
were prepared and tested with a cone calorimeter at 35 kW m2 heat flux. The
results showed that the burning behaviour of PP was significantly reduced in
the presence of fibres. The composites ignited earlier than the PP sample, but
peak and total heat release values were decreased, deceasing further with
increasing fibre content. Important results are shown in Table 5.7.56
5.4.3.2 Thermoplastic Composites Reinforced with Textile

Structures
In our recent research work we have prepared a number of composite samples
using PP and PLA as matrices and jute, sisal, wool and glass fabrics as rein-
forcements. The reinforcement was in a woven fabric form. The area densities
of the reinforcing fabrics varied according to the fibre type used and are
indicated here: wool ¼ 172 g m2, jute ¼ 174 g m2, sisal ¼ 62 g m2 and
glass ¼ 280 g m2. An assembly of each type of nonwoven mat of thermoplastic
fibres and eight layers of a woven reinforcing fabric was melt pressed in a
Table 5.7 Cone calorimetric results for flax/PP composites at 35 k W m2

heat flux.56
Fibre content
in PP/flax TTI PHRRa THR EHC Average MLR
composite (s) (kW m2) (MJ m2) (MJ kg1) (g s1 m2)
0 62 1200 156 41.6 17.9
12.5 45 480, 750 161 39.2 13.7
20 45 470, 560 165 37.2 11.7
30 39 440, 380 125 34.2 9.8
40 35 290, 380 109 33.1 9.2
PHRR values are approximate, calculated from graphs reported by Helwig and Paukszt.56
a
mould at 190 1C for 2.5 minutes, followed by quick cooling to obtain a com-
posite sample sheet. The fibre/polymer ratio for each composite was kept as
40:60 by weight. The details of the composite fabrication technique are pre-
sented in our forthcoming publication.57 Since all of the fabrics were of dif-
ferent area densities, the samples prepared were of different thicknesses, as
shown in Table 5.5. However, for each fabric type the PP and PLA composites
were of similar thicknesses. All of these samples were tested for flammability
in a cone calorimeter at 35 kW m2 external heat flux. Since all samples are of
different thicknesses and the thickness of a sample affects its burning behaviour,
samples of similar thicknesses (produced from fabrics of similar area densities)
are grouped together in Table 5.5. In Figure 5.4, HRR, mass loss and smoke
production versus time curves for all samples of each type of matrix are shown.
All fibres had a minimal effect on the TTI of PP or PLA matrices. All types
of reinforcement reduced the PHRR of PP from 1699 kW m2 to 506–680 kW m2,
PP PLA
1800 700
1500 600
PP PLA
HRR, kW/m2
500
HRR, kW/m2
1200 Gl/PP
400 Gl/PLA
Jute/PP
900 Jute/PLA
Sisal/PP 300
Wool/PP Sisal/PLA
600
200
300 100
0 0
0 100 200 300 400 0 100 200 300 400
(a) Time, s (d) Time, s
100 PP 100
Gl/PP PLA
80 Jute/PP 80
Gl/PLA
Sisal/PP Jute/PLA
Mass, %
Mass, %
60 Wool/PP 60 Sisal/PLA
40 40
20 20
0 0
0 100 200 300 400 0 100 200 300 400
Time, s Time, s
(b) (e)
25 1
20 0.75 PLA
PP
Smoke, m2/m2
Gl/PP Gl/PLA
0.5
Smoke, m2/m2
15 Jute/PP Jute/PLA
Sisal/PP 0.25 Sisal/PLA
10 Wool/PP
0
5 0 100 200 300 400
–0.25
0
0 100 200 300 400
–0.5
Time, s Time, s
(c) (f )
Figure 5.4 (a, d) HRR, (b, e) mass loss and (c, f ) smoke vs. time curves for glass,
jute, sisal and wool fibre-reinforced PP (a–c) and PLA (d–f) composites
exposed to 35 kW m2 heat flux.
104 Chapter 5
which is a 60–70% reduction. From all these fibres, glass displayed the greatest
reduction, followed by wool and then jute and sisal. This trend is as expected from
the flammability of these fibres. However, the THR increased for all natural fibres.
The low value of the THR shown by the sisal/PP composite is due to its lesser
thickness than other samples (Table 5.5). As discussed earlier in Section 4.2, ther-
mally and physically thin samples ignite early, burn for a shorter time and produce
less total heat release compared to thermally and physically thick samples which
burn slowly but for a longer time.51,52 Natural fibre-reinforced composites burn
slowly but for a longer time, producing more total heat and smoke. This effect is
more pronounced in wool/PP composites.
In the case of PLA composites, glass fibres helped in reducing the PHRR by
58% in comparison to pure PLA, jute by 40% and sisal by 18%. This reduction
is much less than that seen for PP composites. The THR for the jute/PLA
sample is increased, whereas smoke production in all samples is minimal.
On comparing the results for PP and PLA composites, the difference between
any one type of fibre-containing composite was similar to that seen for the
two matrices in Figure 5.3. However, owing to different sample thicknesses, the
results cannot be directly compared. In a subsequent publication,56 these results
will be analysed in more detail.
5.4.3.3 Thermoset Composites

The only work reported on the flammability of natural fibre/reinforced ther-
moset composites is by Manfredi et al.14 Composite samples tested include jute,
flax, sisal and glass fibre reinforced unsaturated polyester and modified acrylic
(modar) resins. These samples were prepared by Rodriguez29 by placing fabric
or mats in a mould, impregnating with resin by a vacuum infusion technique
and curing at 60 1C for 2 h and post curing at 110 1C for 3 h. Jute/unsaturated
polyester composites showed the best results on flexural and tensile strengths
and the lowest in impact energy, because of the strong interphase developed.
Flax composites showed higher impact energy than the other natural fibre
composites, due to the existence of the effective energy dissipation mechanisms,
such as pull-out and axial splitting of the fibres.
Manfredi et al.14 studied the flammability by cone calorimeter at 35 kW m2
external heat flux and, from the data reported, the TTI and THR values are
given in Table 5.8. The TTI depends upon the ease of ignition of the material on
the surface, i.e. resin-rich surfaces show low TTI and high peak heat releases,
whereas the THR depends upon the amount of flammable material in the
composite and the duration of burning. From the results in Table 5.8 it can be
seen that there is no real trend of results which can clearly show the effect of
different reinforcing fibres or matrices. To assess the fire risk, the total heat
evolved was plotted against the peak of the heat release rate divided by the
TTI.29 Among the composites with an acrylic matrix, the one reinforced with
sisal fibre showed the highest fire risk and in global terms the worst fire resis-
tance. Jute fibre composites showed a quick growing but short duration fire
and, conversely, flax fibre composite developed a long duration but slow
Flame Retardant Characteristics of Natural Fibre Composites
Table 5.8 Mechanical and cone calorimetric results at 35 kW m2 heat flux for natural fibre reinforced polyester and acrylic
composites.14,29
Mechanical properties Cone results at 35 kW m2
Flexural Tensile Impact Water
modulus modulus energy absorp. PHRRb THR Av. SEA
Sample a
Reinforcement type (GPa) (GPa) (kJ m2) (%) TTI (s) (kW m2) (MJ m2) (m2 kg1)
Jute/acrylic Woven jute fabric 4.6 6.9 8.8 3.0 72 520, 910 74.2 225.5
Jute/polyester Woven jute fabric 6.6 8.0 10.6 2.7 51 395, 515 77.6 736.1
Sisal/acrylic Nonwoven mat 2.5 3.4 12.7 3.2 52 310, 470 101.4 432.9
Sisal/polyester Nonwoven mat 3.9 5.3 12.2 2.7 – – – –
Flax/acrylic Nonwoven mat 5.9 6.3 15.0 2.8 110 580 104.9 460.7
Flax/polyester Nonwoven mat 4.8 6.3 13.2 – – – – –
Glass/acrylic Nonwoven mat 11.3 13.3 98.7 0.36 62 340, 310 54.4 491.6
Glass/ Nonwoven mat 11.4 14.9 106.5 0.4 56 275 32.3 709.5
polyester
a
All composites had 30% fibre by volume; sample thickness ¼ 3–4 mm.
b
PHRR values are approximate, calculated from graphs reported by Manfredi et al.14
105
106 Chapter 5
growing fire. Glass fibre composites showed more fire resistance than the nat-
ural fibre composites. Comparing the different matrices, it was observed that
the fire risk was similar between the composites with unsaturated polyester and
acrylic matrices. The jute fibre composites released lower quantities of smoke
than the other composites (see Table 5.8). Comparing the different matrices,
unsaturated polyester composites produced more smoke than the acrylic
composites.
5.5 Flame Retardant Strategies for Components/

Composites
Since composites are composed of two or more components, any one or both of
these can be rendered flame retardant prior to making a composite structure.
Alternatively, the finished composite structure can be surface treated with
protective coatings. These are discussed here as three generic methods of ren-
dering the composites flame retardant:
5.5.1 Flame Retardant Matrices: Use of Reactive or Additive

Flame Retardants
A matrix can be flame retarded by adding flame retardant chemicals into it or
by chemical modification of the polymer backbone by introducing flame
retardant elements into it. Alternatively, an inherently flame retardant matrix
can be used, e.g. PVC for thermoplastics and phenolics for thermosets. While
additives can be used for both thermoplastics and thermosets, chemical mod-
ification of the resin is only feasible in thermosets due to a vast choice of resins,
curing agents and hardeners. For example, in unsaturated polyesters the use
of halogenated resins or replacement of the curing agent from styrene to bro-
mostyrene, introduction of halogen elements such as chlorine into the epoxy
backbone in the diglycidyl ether of bisphenol C (DGEBC), fluorine in the
diglycidyl ether of bisphenol F (DGEBF) or bromine in tetrabromobisphenol
A (TBBA). For a detailed discussion on flame retardancy of thermosets, the
reader is referred to previous reviews.8,9
The use of additives is the most popular method of flame retarding ther-
moplastic composites containing short fibres, where flame retardant (FR)
chemicals can be added into the polymer and fibre prior to melt compounding.
For long fibre composites, FR chemicals can be introduced into the polymer
prior to forming sheets. The choice of flame retardant depends upon the type
and processing temperature of the polymer used in the composite. There are a
number of literature reviews available on flame retardants for different polymer
types.58–63 Most commonly used flame retardants (as reported in literature) for
thermoplastic polymers include aluminium hydroxide, magnesium hydroxide,
expandable graphite, ammonium polyphosphate, intumescent systems and
nanoclays. Metal hydroxides such as aluminium hydroxide and magnesium
hydroxide act as flame retardants by undergoing endothermic reactions at
elevated temperatures, reducing the polymer temperature and releasing water
vapour, which effectively dilutes the volatile species emanating from the
polymer degradation. However, metal hydroxides are effective only at very
high percent loadings (typically 450 wt%), which might have an adverse effect
on the mechanical properties of the polymer matrix. Nitrogen- and phos-
phorus-containing additives such as ammonium polyphosphate, intumescent
chemicals, etc., on the other hand, chemically interfere with the polymer
decomposition and promote char formation. Chemically reactive type flame
retardants are required at comparatively lower levels (20–30 wt%). Some spe-
cific examples of the use of these flame retardants in natural fibre composites
are discussed here.
Hapuarachchi et al.64 have reported the use of ATH (alumina trihydrate or
aluminium hydroxide) (40 wt%) to flame retarded hemp/unsaturated polyester
composites. They showed that ATH increased the TTI from 54 s in the control
to 78 s and reduced the PHRR from 361 to 176 kW m2, when subjected to
50 kW m2 external heat flux. Sain et al.65 have used magnesium hydroxide on
sawdust and rice husk-filled polypropylene composites. Replacement of 25% of
the natural filler with magnesium hydroxide reduced the horizontal burning
rate (ASTM D 635) by around 50%. No synergetic effect was observed when
magnesium hydroxide was used in combination with boric acid and zinc borate.
Schartel et al.36 have compared the effectiveness of ammonium polypho-
sphate and expandable graphite as flame retardants for flax/PP (40:60 by wt%)
composites containing short fibres, prepared by melt blending all components
together. Thermogravimetry in a nitrogen atmosphere was used for studying
the pyrolysis behaviour, whereas flammability was studied by LOI, UL94,
the glow wire test and cone calorimetry at different heat fluxes ranging from
30 to 70 kW m2. Selected results extracted from their paper are presented in
Table 5.9. Both flame retardants promoted char formation, as observed by
thermogravimetry and cone results. Ammonium polyphosphate and other
(mono- and diammonium) phosphates on heating decompose to phosphoric
acid, which esterifies the hydroxyl groups of the cellulosic structure of natural
fibres. The resulting cellulose ester increases the char amount of the decom-
position.66 Polyphosphoric acid does not chemically react with decomposing
PP as PP undergoes decomposition via random chain scission involving for-
mation of some unsaturated end groups, e.g. –C(CH3)¼CH2, which on oxi-
dation yields further scission products and has no tendency to cross-link and
form char.67 Expandable graphite undergoes endothermic decomposition and
expansion up to 300 times its initial volume under the impact of heat. The
expanded charred structure creates a heat insulating layer for the underlying
substrate, hence imparting flame retardancy. All flax/PP composites showed
two peaks for the heat release rate in the cone calorimeter: the first one at the
beginning of burning and the other one at the end, when the char cracks on the
thermal feedback of the sample holder. The double peak behaviour is due to the
lignocellulosic content of flax fibres, similar to the behaviour known for
wood.68 In flame retarded samples the first peak is reduced significantly due to
char formation, which reduces the burning propensity of the composite. The
108 Chapter 5
Table 5.9 Flammability results for flax/PP composites containing APP and
expandable graphite.36
Cone resultsa at (35) and 50 kW m2
Critical Char
TGA PHRR THR HRRb residue
Sample and mass residue at LOI (kW (MJ (kW at flame-
composition 490 1C (vol%) UL94c m2) m2) m2) out (%)
PP/flax (70:30) 13.9 21 HB (410), (130), 167 10.6
520 135
PP/flax/APP 27.4 26 HB (210), (95), 131 29.3
(45:30:25) 270 95
PP/flax/expandable 29.4 30 V1 (130), (85), 35 429.6
graphite (45:30:25) 160 95
a
The values are approximate, extracted from a figure in Schartel et al.36
b
Calculated by extrapolating the steady heat release rate obtained from different heat fluxes in the
cone calorimeter to an external heat flux of zero.
c
HB ¼ the sample was tested in a horizontal position and found to burn at a rate less than a specified
maximum.
V1 ¼ the sample was tested in a vertical position and self-extinguished within a specified time after
the ignition source was removed.
values extracted at 35 and 50 kW m2 external heat fluxes in Table 5.9 show
that expandable graphite is more effective in reducing both peak and total heat
release values compared to APP (ammonium polyphosphate). Critical heat flux
values, obtained by extrapolating the steady heat release rate obtained from
different heat fluxes in the cone calorimeter to an external heat flux of zero, also
indicate the efficacy of expandable graphite compared to APP. This is further
supported by LOI and UL94 results, where only expandable graphite-
containing samples could achieve a V1 rating, whereas both control and APP-
containing samples failed the test.
In a similar study, Le Bras et al.54 studied the effect of APP on the pen-
taerythritol/melamine system. Flax/PP (40:60 by wt%) composites were pre-
pared by extruding PP and short flax fibres (20 mm mean length). On adding
APP to the sample as PP/flax/APP (31:46:23 wt%), the TTI, PHRR and THR
values decreased. Even with a decrease in PHRR from 640 to 300 kW m2 the
sample could not pass the UL94 test due to a flaming time higher than 30 s and
burning drops. However, when APP was replaced by an intumescent system
[flax/PP/(APP/pentaerythritol/melamine), 31:46:23 wt%], the sample showed
the formation of a stable intumescent char with no polymer dripping and
instantaneous complete extinction after the second flame exposure, obtaining
V0 rating. It shows that char-forming fibres need an intumescent composite
structure maintaining coherence of the protective shield and thermal barrier
properties to give a high level of flame resistance to the material. However,
in the cone calorimetry test the intumescent system did not show any further
improvement in PHRR (270 kW m2) and THR values.
Matko et al.55 studied the efficiency of diammonium phosphate and
ammonium polyphosphate in polypropylene, polyurethane and starch-based
biocomposites. Natural fibre reinforcement was in the form of wood flake
(pinewood as sawdust, assorted of 1.2 mm) or corn shell (3–12 mm size). APP
was added to the polymer mix during polymer processing, whereas diammo-
nium phosphate was used to treat the lignocellulosic fillers by mixing two
components in 1:1 ratio in aqueous solution and then drying the material under
an infrared lamp. Polypropylene biocomposites were prepared by compound-
ing different components in a Brabender mixer and then compression moulding
into sheets. Polyurethane biocomposites were prepared by mixing the wood
flake/corn shell fillers and flame retardant into the mixture of polyol and an
isocyanate at room temperature, followed by pressing into sheets. Starch
samples contained corn starch and glycerol, as plasticizing agent, in a 1:1 mass
ratio and required amount of flame retardant and compounding was carried
out in a Brabender compounder. Composition and flammability results for
these samples are summarized in Table 5.10. As can be seen from the results
in the table for wood flake/PP composites, the sample containing 10% wood
flakes and 10% APP had a LOI of 22 vol%, which is higher than that of the
pure PP (LOI ¼ 18 vol%); the value, however, hardly changed between 10 and
40% wood flake content. On increasing the fibre content up to 50% the LOI
reached 30 vol% and achieved the V0 rates of flame retardancy. Hence, the
combination of wood flake reinforced PP composite with APP shows flame
retarded character only at high concentration of reinforcing component. The
20% APP containing samples were slightly better, with LOI reaching 24 vol%
with 10% wood flakes and 32 vol% with 50% wood flakes.
Table 5.10 Flammability results of wood flake and corn shell containing
polypropylene (PP) and polyurethane (PU) composites and their
flame retarded counterparts.55
Composite Flame retardanta LOI (vol%) UL94b
PP þ wood flake (10–40%) APP (10%) 22–25 HB
PP þ wood flake (10–40%) APP (20%) 24–27 HB
PP þ wood flake (50%) APP (10%) 30 V0
PP þ wood flake (50%) APP (20%) 32 V0
PU (80%) þ wood flake (20%) – 23 HB
PU (60%) þ wood flake (20%) APP (20%) 31 V0
PU (60%) þ wood flake (20%) DAP (20%) 30 V0
PU (80%) þ corn shell (20%) – 20 HB
PU (60%) þ corn shell (20%) APP (20%) 27 V0
PU (60%) þ corn shell (20%) DAP (20%) 29 V0
Corn starch þ glycerol (1:1) – 23 NR
Corn starch þ glycerol (1:1) APP (5%) 28 HB
Corn starch þ glycerol (1:1) APP (10%) 33 V0
a
DAP ¼ diammonium phosphate.
b
HB ¼ the sample tested in a horizontal position and burned at a rate less than a specified max-
imum; NR ¼ No rating; V0 ¼ the sample was tested in a vertical position and self-extinguished
after the ignition source was removed.
110 Chapter 5
For PU composites, both samples containing wood flakes or corn shell had
low LOI values, i.e. 23 and 20 vol%, respectively. However, adding 20% of
APP increases them to a great extent (to 31 and 27 vol%, respectively). A V0
rate was achieved with all the flame retarded systems. The LOI value of plas-
ticized thermoplastic starch is quite low; the introduction of a small amount of
APP (10%) increased the flame retardancy up to the self-extinguishing V0
rating. On increasing the concentration of APP, a sample with LOI of 60 vol%
could be obtained. The outstanding flame retardancy results of starch-based
thermoplastic biopolymer systems, which outperform the other studied bio-
composites considerably, can be explained by the polyol character of the
polymer matrix and the presence of polyol plasticizer. Thus the flame retar-
dancy of such biopolymer systems can be solved quite easily by introducing
phosphorus charring components. From this study it can be observed that that
both the polysaccharides and polyurethane act as a charring agent and their
content in the biocomposites correlates well with the achievable flame retar-
dancy level. Plasticized starch thermoplastic, a fully degradable biopolymer,
can be flame retarded efficiently by the introduction of as low as 10% APP and
a LOI ¼ 60 vol% value can be achieved with 30% flame retardant additive.
Generally, it can be concluded that although bio-based polymers seem to be
more expensive than the traditional mass polymers, their flame retardancy can
be achieved in relatively simple and inexpensive ways.
Since the 1990s, polymer nanocomposites obtained by the addition of
nanoparticles such as organoclays, nanosilica and carbon-based nanotubes into
polymers have attracted considerable interest due to their ability to improve the
physical and mechanical properties of polymeric materials while also enhancing
their thermal stability and flammability. Although the use of nanoclays is vastly
reported for bulk polymers69 such as polypropylene,66,67,70 polystyrene,69
unsaturated polyester71 and epoxy resins,41 no specific examples for natural
fibre-reinforced composites could be found in the literature.
5.5.2 Treatment of Natural Fibres with Flame Retardants

For cellulosic fibres a number of phosphorus- and nitrogen-based flame
retardants (phosphates, phosphoramides, etc.) are available. Many reviews
have been published on the flame retardancy of natural fibres/textiles.66,72–75
Most of the flame retardants used for textile applications can also be used on
the fibres/fabrics prior to composite formation, provided that the decomposi-
tion temperature of the flame retardant is higher than the composite’s pro-
cessing temperature. Candidate flame retardants are ammonium phosphates
such as mono- or diammonium phosphates, ammonium bromide in combi-
nation with ammonium phosphates to provide some vapour-phase flame
retardant action, borax and boric acid, ammonium sulfamate and sulfates.
Durable flame retardant finishes used for cotton fabrics can also be applied.
Examples include N-methylol dialkyl phosphonopropionamides, from
which a well-known product, Pyrovatex CP (Ciba), is derived and tetra-
kis(hydroxymethyl)phosphonium derivatives [Proban CC (Rhodia, previously
66
Albright & Wilson)]. Most of these flame retardants function in the con-
densed phase and promote char formation by converting the organic fibre
structure into carbonaceous char and hence reduce volatile fuel formation.11
These condensed-phase active flame retardants are very effective in natural
fibres owing to the polymer backbone having reactive side-groups, which on
removal lead to unsaturated carbon bond formations and eventually a carbo-
naceous char following elimination of most of the non-carbon atoms present.11
Vapour-phase active halogenated flame retardants are also very effective on
cellulosis, which function in the vapour phase by interfering with the flame
chemistry through free radical generation. The detailed mechanisms of action
of these flame retardants on textiles are similar to the ones in bulk polymers, for
which readers are referred to other reviews.58–60
Natural fibres/fabrics are generally treated with flame retardant finishes
in the finished fabric form. Finishes are usually applied by a pad-dry
method where the fabric is passed through the chemical formulation (mostly
in aqueous form), passed through rollers to squeeze out the excess and
then dried in an oven at 120 1C. This gives a non-durable finish. To obtain
a semi-durable or durable finish, the fabric is passed through another oven
set at a higher temperature (usually 160 1C), where a curing stage allows a
degree of interaction between the finish and the fibre. These treatments can
also be applied to nonwoven webs of fibres or short/long fibres by spraying
the flame retardant finish and then drying the fibres prior to composite
formation.
Intumescent chemicals can also be used for treating the fibres/fabrics. Intu-
mescent coatings have been around for nearly 70 years, normally used as
surface treatments for structural materials and metals to protect them from
heat and fire. On heating, they form a foamed char, which thermally insulates
the underlying structure. In textiles, although they can be used a surface coating
on the fabric, they are usually applied in-between different fabric layers, which
are then needle-bonded to consolidate the whole structure.66 In our work on
flexible composites76 we applied an aqueous formulation of intumescent che-
micals and a binder on a nonwoven web using a brushing technique. After
drying, a second nonwoven web was attached to it and the whole assembly was
then needle punched into a coherent single web.76 In a later study the intu-
mescent formulation was applied by padding on the nonwoven web.77 We have
demonstrated that intumescent chemicals can chemically interact with cellu-
losic fibres and this results in high levels of flame and heat resistance.76,78 We
also have shown that on introduction of these intumescent/flame retardant fibre
systems to thermoset (epoxy, polyester and phenolic) resins, there is a physical
and chemical interaction of the three components leading to enhanced char
formation.79 Glass fibre reinforced rigid composite structures containing these
intumescent/flame retardant cellulosic fibres have shown superior flame retar-
dant properties without any detrimental effect on their mechanical proper-
ties.51,77,79,80 The intumescent/FR fibre combinations may be introduced either
as a pulverized additive to the resin or as an additional textile fabric layer to the
composite structure.77
112 Chapter 5
With regards to specific examples in natural fibre composites, this metho-
dology of flame retarding composites has not much been exploited. The only
example available is by Misra et al.,30 where tin(II) chloride treatment was used
for brominated coir fibre in an epoxy-based composite. The coir fibre was
treated with saturated bromine water for increased electrical properties and
then with tin(II) chloride solution to increase the fire resistance. The fibre was
ground to a powder and then mixed with epoxy resin to make the composite.
They found that a 5% tin-based halogen compound reduced the smoke density
by 25% and increased the LOI value from 36 vol% in a control to 39 vol% in
the FR composite.
5.5.3 Surface Treatments of Composites

The most efficient way to protect materials against fire without modifying their
intrinsic properties (e.g., mechanical properties) is the use of fire retardant
coatings.81 In the case of glass/carbon fibre-reinforced thermoset composites,
thermally insulative surface coatings can protect the resin-rich surface from
ignition. This means of protecting flammable materials is called ‘‘passive fire
proofing’’, as it serves to decrease heat transfer from the fire to the structure.
Most of the effective coatings are ceramic- or intumescent-based. Intumescence
is defined as the swelling of a substance when exposed to heat, typically forming
a multi-cellular, carbonaceous or ceramic layer, which acts as a thermal barrier
that effectively protects the substrate against a rapid increase of temperature,
thereby maintaining its structural integrity.82 The alternative is to use ceramic
fabrics, ceramic coatings, hybrids of ceramic and intumescent coatings, silicone
foams or a phenolic skin, all of which showed good performance for glass-fibre
reinforced thermoset composites.83 Use of mineral and ceramic claddings is
quite popular for naval applications to fireproof conventional composite
structures. These barriers function as insulators and reflect the radiant heat
back towards the heat source, which delays the heat-up rate and reduces the
overall temperature on the reverse side of the substrate. One commercial
example of this sort of product is the Tecnofire range of ceramic webs produced
by Technical Fibres in the UK; these are available with a number of different
inorganic fibres, including glass and rock wool either with or without an
associated exfoliated graphite present. They are designed to be compatible with
whatever resin is used in composite production. We have shown their effec-
tiveness for glass fibre-reinforced polyester composites.84,85 3 M Nextel 312
woven fabrics from alumina/boria/silica fibres are also used as composite fire
barriers. Phenolic foams are also very effective fire barriers and particularly
used for bulkhead structures in military ships.
There is no example of usage of the above-mentioned surface barrier coat-
ings/ceramic claddings for natural fibre composites available in the literature,
either for thermoset or thermoplastic composites. While surface treatment can
be easily used for thermoset composites, the problem with thermoplastic
composites could be that on nonpolar matrix surfaces the polar intumescent
chemicals or inorganic/ceramic coatings do not stick. Use of compatiblizers,
however, can solve this problem.
5.6 Summary
Natural fibre composites are becoming popular in the automotive industry
owing to their renewability, biodegradability and cost. The use of natural
fibres in a composite reduces the flammability of the composite compared to
the pure polymer. However, for these composites to be used in other sectors,
such as marine, aerospace or construction, they will have to pass commercial
fire tests, such as UL94, flame spread, etc. For this, some sort of flame
retardant treatment is required. Flame retardants can be added into the
thermoplastic polymer or the natural fibre textile reinforcement can be flame
retarded using conventional textile finishing processes. The composite can
also be surface treated with flame retardant chemicals such as intumescent
paints.
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66. B. K. Kandola, in Fire Retardancy of Polymeric Materials, ed. A. B.
Morgan and C. A. Wilkie, CRC Press, New York, 2010, pp. 725–761.
67. B. K. Kandola, A. Yenilmez, A. R. Horrocks and G. Smart, in Fire and
Polymers, ed. A. Morgan, G. L. Nelson and C. A. Wilkie, ACS Symp. Ser.
1013, American Chemical Society, Washington, 2009, pp. 47–69.
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Adv. Technol., 2008, 19, 693–700.
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35, 143–155.
CHAPTER 6
Natural Fibre Composites:

Automotive Applications
S. C. R. FURTADO,a A. J. SILVA,a C. ALVES,*b
LUÍS REIS,a MANUEL FREITASa AND PAULO FERRÃOa
a
Instituto Superior Técnico, Avenida Rovisco Pais 1049-001, Lisbon,
Portugal; b Universidade Federal do Rio Grande do Norte, Campus
Universitário - Lagoa Nova, Av. Sen. Salgado Filho, 3000, 59078-970, Natal,
RN, Brazil
6.1 Introduction
Composite materials have been used by humans for some millennia. Presently,
the most common polymer matrix composites, able to carry significant loads,
use carbon, aramid (aromatic polyamides), or glass fibres as reinforcements.
Significant contributions can be found in the literature regarding their
mechanical behavior in static and dynamic conditions, their ageing behavior
and, more recently, their environmental impact.
Industry in general is looking for ‘‘greener’’ materials (from renewable sources)
to use in its products, both as a way of reducing the environmental burden and as
a way of appealing to a growing environmentally conscious market. The auto-
mobile industry in particular is constantly looking for products that are lighter,
more eco-friendly, and are still adequate for mass production at low cost.
A tentative shift from steel to light alloys and composite materials in the later
part of the last century was immediately followed by a reaction from the steel
industry with the introduction of high-strength steels and other special alloys
that could also reduce the weight of products. The major focus of the steel

118
Natural Fibre Composites: Automotive Applications 119
industry is the recyclability of steel, compared with the recyclability of other
metal alloys or composite materials.
One advantage of using natural fibres such as kenaf, hemp, flax, jute, and
ramie, in substitution of aramid or glass as fibre-reinforced composites, is their
renewable nature and inherent biodegradability, rendering the issue of reuse or
recyclability meaningless. Using natural fibres in automotive structures (espe-
cially in enclosures) could be an advantage over other non-recyclable reinforced
polymers, provided that their mechanical behavior is fully understood and
comparable with more traditional solutions.
Auto companies are seeking materials that combine different properties, like
sound abatement capability and reduced weight. It is estimate that B75% of a
vehicle’s energy consumption is directly related to factors associated with the
vehicle’s weight.1,2 To improve the fuel economy, manufacturers have focused
on reducing automobile weight, primarily through manufacturing methods that
use raw materials offering the strength of steel without its heft.1 Composites
reinforced by vegetable fibres, like jute, flax, hemp, or ramie, can be a good
choice. It was shown that they can be a realistic alternative, for example, to
glass-reinforced composites, because they can deliver the same performance for
lower weight, can be 25–30% stronger for the same weight, and can exhibit
favorable non-brittle fracture on impact, which is an important requirement in
the passenger compartment.2 According to BMW, it is possible to manufacture
bio-based composites that are as much as 40% lighter than equivalent injec-
tion-molded plastic parts.3
Environmental legislation in different countries is becoming much more
stringent, leading to an increased use of natural fibre-reinforced plastics in
automotive construction and pushing the automakers to evaluate the envir-
onmental impact of a vehicle’s entire lifecycle, from raw materials to manu-
facturing to disposal.4 A regulation example is the European Directive 2000/53/
EC, in which vehicles have to be partially decomposable or recyclable (95%) by
2015.3,5 Japan also requires that 95% of a vehicle be recovered (which includes
incineration of some components) by 2015.3 In the USA, 10–11 million vehicles
reach the end of their useful lives each year. A network of salvage and shredder
facilities process about 96% of these old cars; about 25% of the vehicles weight,
including plastics, fibres, foams, glass, and rubber, remains waste. A car made
mostly of heated, treated, and molded bio-fibre would simply be buried at the
end of its lifetime and consumed naturally by bacteria,2 leading to less necessity
of landfill.
The use of natural composites in the automotive industry started with Henry
Ford. Around 1942, he began experimenting, initially using compressed soy-
beans to produce composite plastic-like components (Figure 6.1). During that
period the petroleum-based chemicals were very cheap, and therefore soy-
based plastic could not find economic importance. With the new environ-
mental regulations and depletion and uncertainty of petroleum sources,
scientists and manufacturers have revived their interest to derive a new
generation of composite materials from renewable sources like soybean-based
plastic or natural fibre.6 Many major automakers have now a real interest in
120 Chapter 6
Figure 6.1 Henry Ford performing a durability test with a fire axe on a prototype car
made of plastic derived from soybeans.6
Table 6.1 Some fibre prices; vegetable fibre prices as of 2008;10 E-glass fibre
price for 2011.11
Fibre Flax Hemp Jute Abaca Sisal Coir E-glass
Price (US $/kg) 0.747 0.754 0.332 1.155 0.825 0.314 2
global sustainability. DaimlerChrysler identifies bio-based materials as one of

the two key parts of its plan to create a global sustainability network.7
Over the last few years, a number of researchers have been involved in
investigating the exploitation of natural fibres as load bearing constituents in
composite materials.8 In Europe, automotive companies have developed
components made of vegetable composites, motivated mainly by regulations
that have played an important role toward sustainable mobility. In the USA,
about 1.5 million vehicles are already using vegetable fibres as reinforcement
for thermoplastic and thermosetting polymers. In Brazil, some automotive
initiatives are concerned with the selection of ‘‘greener’’ materials (from
renewable sources). Several studies point out that automotive textile usage has
grown in terms of quantity, quality, and product variety worldwide.5
The use of such materials in composites has increased owing to their relative
cheapness (Table 6.1), the fact that surface treatments avoid the use of organic
solvents, and that they can compete well in terms of strength per weight of
material.2,8 They also contribute to a reduction in fossil fuel consumption and
greenhouse gas (GHG) emissions of vehicles by consuming about 45% less
energy in their incineration, which complies with environmental directives and
the Kyoto Protocol.5 NPSP Composieten have introduced bio-composites in
external and internal body panels for trains and train furniture for the Dutch
Railways, which resulted in a lower train weight (1000 kg per train), saving
5 MWh of electricity per train per year, which led to less pollution of about
0.5 metric tons of CO2 a year.9
Automakers now see strong promise in natural fibre composites.2,5 Targeting
the environmental requirements of vehicles warrants a shift of vehicle design
from oil-derived polymers and synthetic reinforcements to natural materials.
Undoubtedly, vegetable or ‘‘green’’ composites are playing a major role in
the product design process, presenting many environmental advantages com-
pared to oil-based composites, and offering a possibility to developing countries
to use their own natural resources in a wide range of industries, as well as
resulting in social aspects such as new jobs in impoverished local communities.5
Currently, at least 315 000 metric tons of bio-composites reinforced by vege-
table fibres are already being used in the European industry, mainly in the
automotive and construction sectors. By 2020, this quantity could be more than
doubled.12
6.2 Composite Materials: Definition and Classification

Composites are one of the oldest materials used by humans, and there have
been significant developments due to the booming of the aviation industry in
the mid-20th century plus the emergence of the aerospace industry. Since then,
many investments have been used to develop and produce polymeric resins and
glass fibres, which motivated their use widely, with glass fibres becoming the
main composite reinforcement worldwide.13 Several industrial applications
require properties that are not found in neat materials; thus composite mate-
rials can be feasible to meet industrial requirements presenting useful
mechanical properties through system synergy, in which an applied load stress
is supported by the matrix and reinforcement. In this sense, it is clear that the
interface reinforcement/matrix plays an important role in composites, influ-
encing their final properties. Besides the mechanical advantages of composite
materials, they also give a great advantage for design processes, allowing
designers to assume a role beyond merely ‘‘material picker’’.14 Therefore, the
success of a product made of composite materials will depend on the best
designer choice related to the matrix/reinforcement combination, besides other
aspects such as cost and production.15
According to the American Society for Testing and Materials (ASTM),
composite materials are the combination of two or more different materials to
form a new and useful engineering material, differing from the neat materials
that originated them. They can be produced by combining metals, ceramics, or
polymers,16 and their constituent elements should be distinguished on a mac-
roscopic scale. Most composites have better mechanical behavior than these
phases or even new properties that emerged from their combination.17,18
According to Jones,17 some of the main improved properties are strength,
hardness, fatigue and corrosion resistance, weight, and thermal conductivity.
122 Chapter 6
Generally, composites are made mainly of two different materials, well
known as matrix and reinforcement. The matrix of a composite performs
essential functions such as to protect the reinforcement from the environment,
ensure an efficient distribution of an applied load stress to the reinforcement,
and to provide a stable shape of the composite. On the other hand, the rein-
forcement performs the structural function, increasing one or more properties
of the composite.
6.2.1 Matrices
Matrices can be distinguished as metallic, ceramic, or polymeric. The polymeric
ones are widely used to produce components for various industrial markets,
since they present lower costs than other matrices. They also present low
density, which implies lightweight composites. Polymeric matrices are divided
into two categories: thermoplastic and thermoset. Thermoplastic matrices are
composed of large molecules bound to each other by van der Waals and
hydrogen bonds. When heated, these bonds are temporarily broken, allowing
molecular mobility; after cooling the molecules assume new positions,
reshaping the material. Usually, thermoplastic polymers have higher impact
strength and toughness than thermosetting polymers. They are easily processed
by injection or thermoforming and can be recycled, which is an important
behavior related to their environmental performance. Nowadays, they repre-
sent about 35% of the global composite market and the thermoplastic polymers
most used are the following: polypropylene, polyamide, polycarbonate and
PEEK (polyether-ether-ketone).19 Thermosetting matrices are composed of
three-dimensional molecules, forming rigid and permanent structures that do
not allow their reprocessing and/or reshaping. Generally, they are processed by
adding accelerator and catalyst into the resin to start the polymerization,
forming a three-dimensional structure in a process called ‘‘curing’’. Worldwide,
the most used thermoset matrix to manufacture industrial composites is the
unsaturated polyester, mainly due to its low cost, good properties, and low
viscosity, which implies better wettability and impregnation of the reinforce-
ment. These matrices can also be divided into three groups: orthophthalic for
general industrial usage, isophthalic that presents good thermal behavior, and
bisphenol used in aggressive environments.20 Table 6.2 shows some thermo-
plastic and thermoset resins as well as some characteristics.
6.2.2 Reinforcements: Fibres and Fillers

In composite materials, reinforcements are the components which improve
their structural properties as a whole and support most load stress, with a
percentage also influencing the final cost of the composites. More than one
type of reinforcement can be used in a composite to obtain the intended
properties, forming a combined composite (Figure 6.2f). Most reinforcements
have large length/diameter ratios (fibre form), which are widely accepted as
a
Table 6.2 Mechanical properties of thermoplastic and thermoset resins.
Density Young’s modulus Tensile strength
Resin (g m–3) (MPa) (MPa)
Thermoplastic Nylon 1.1 1.3–3.5 55–90
PEEK 1.3–1.35 3.5–4.4 100
PPS 1.3–1.4 3.4 80
Polyester 1.3–1.4 2.1–2.8 55–60
PC 1.2 2.1–3.5 55–70
PTFE 2.1–2.3 – 10–35
Thermoset Epoxy 1.2–1.4 2.5–5.0 50–110
Phenolic 1.2–1.4 2.7–4.1 35–60
Polyester 1.1–1.4 1.6–4.1 35–95
Adapted from Mazumdar.21
a
(a) (b) (c)
(d) (e) (f)
Figure 6.2 Composite classification.
reinforcements for structural and non-structural industrial applications. In

polymeric matrix composites the most used fibres are carbon, aramid, boron,
and glass. Among them, glass fibre is widely used to produce industrial
components, mainly due to its great cost/benefit ratio. Different types of glass
fibres can be produced, based on chemical combinations to meet specific
requirements from different markets. Currently, two types of glass fibres
represent about 90% of the global fibre consumption:13
E-type. This represents more than 90% of total glass fibre usage in the
industrial market owing to its good mechanical and electrical properties
and moisture resistance.
S-type. This is widely used in the construction sector owing to its higher
mechanical properties (about 20–30%) than E-type glass fibre, besides its
124 Chapter 6
better thermal and fatigue resistance. However, its high cost restricts its
industrial usage.
Fillers are another type of reinforcement, which are particles dispersed into
the matrix to achieve the intended composite properties such as cost, weight,
electrical conductivity, mechanical strength, color, and to decrease contrac-
tion.20,22 Usually, fillers are obtained from natural deposition, the most widely
used being calcium carbonate, silica, quartz, graphite, and a large number of
metal powders.23
Nowadays, calcium carbonate (CaCO3) is the mineral most used as filler in
several industrial sectors, such as paper, ceramic, paints, pharmaceuticals, and
plastics. It is also used in environmental applications, e.g. water treatments and
to neutralize acid rain. There are many types of CaCO3 filler, depending on
their origin and their particle treatment, resulting in different composite
properties.22,23 Some filler characteristics such as porosity and dimensions are
very important because they define the system interface, in which the strength
increases as the particle dimensions decrease. Moreover, the dispersion of fillers
into the matrix must be as homogenous as possible to avoid tensile con-
centrations, which decrease the composite mechanical properties.
According to Nezbedová24 and Eylisson,25 the main constraints to filler
usage are related to the properties and compounds of their components, filler
porosity, the required homogenous dispersion of fillers into the matrix, and
the interaction between fillers and matrix. Composite materials are classified
as:17,26
Fibrous composite materials: composed of fibres (reinforcement material

with a large length/diameter ratio) in the matrix. In fibrous composites the
reinforcement can be arranged in different directions: uniaxial (Figure
6.2a), biaxial (Figure 6.2b), and multi-axial (Figure 6.2c). They can also be
divided into short fibre webs (chopped fibre random fleece) and endless
fibres webs (continuous fibre random mat).20
Laminated composite materials: composed of layers from at least two
different materials, which are linked to form a laminate (Figure 6.2d).
Particle composite materials: composed of macroscopic particles (fillers)
of one or more materials, dispersed into matrix (Figure 6.2e).
Combined composite materials: combination of fibrous and particle
composite materials described above (Figure 6.2f).
Composite materials are also classified based on their matrices and/or

reinforcement origins, which can be metallic, ceramic, or polymeric.18
6.3 Natural Fibres

Natural fibres are subdivided based on their origins, coming from plants,
animals, or minerals. All plant (vegetable) fibres are composed of cellulose,
while animal fibres consist of proteins (hair, silk, and wool). Vegetable fibres
Natural
Fibres
Organic Inorganic
Vegetabel Animal Mineral

Fibres Fibres Fibres
Seed-Hair Leaf/Hard Wool/Hair Asbestos

Bast fibres Fruit fibres Spear fibres Silk
fibres fibres fibres ....
• Cotton • Flax • Coir • Bamboo • Wool of • Mulberry

(Gossypium (Linum • Abaca (Cocos (Bambusa sheep • Tussah
(Musa textilis
hirsutum L.,
G. herbaceum
usitatissimum
NÉE)
nucifera L.)
• ....
vulgaris and • ....
L.) SCHRAD, camels:
L., G. • Hemp • Sisal Bambusa alpaka
Barbadense (Cannabis (Agave arundincea llama
L., et al.) sativa L.) sisalana P.) WILLD, and ....
• Akon • Jute • Henequen others) • Rabbit
(Asplecias (Corchorus (Agave • .... hair:
gigantea capsularis L./ fourcroydes angora fibre
R.BR./ corchorus LEM) ....
Calotropis olitorius L.) • ..... • Goat air:
gigantea • Ramie Mohair fibre
NOR.) (Boehmeria Cashmere
• Kapon nivea H. et A.) ....
(Ceiba • Kenaf • Horse
pentandra L.) (Hibiscus hair and
• Paina cannabinus other
(Bombax L.) coarse
ceiba L.) • .... animal
• .... hair
Figure 6.3 Classification of natural fibres.27
include bast (or stem or soft sclerenchyma) fibres, leaf or hard fibres, and seed
fibres.8 Figure 6.3 presents a classification of natural fibres.27
Plant/vegetable fibres are the natural fibres most utilized; because of this, the
expression ‘‘natural fibres’’ is widely used to designate plant/vegetable fibres.
Vegetable fibres can be considered as naturally occurring composites.28 They
are cell walls that occur in stem and leaf parts and comprise cellulose, hemi-
celluloses, lignins, and aromatics, waxes and other lipids, ash, and water-
soluble compounds (Figure 6.4).29 The main components of vegetable fibres are
cellulose (a-cellulose), hemicellulose, lignin, pectin, and waxes.8
The properties of each constituent contribute to the overall properties of the
fibre. The reinforcing efficiency of vegetable fibre is related to the nature of
cellulose and its crystallinity. A high cellulose content and low microfibril angle
are desirable properties in a fibre to be used as reinforcement in polymer
composites. The cellulose fibrils are aligned along the length of the fibre, which
render maximum tensile and flexural strengths, in addition to providing
rigidity.28 Hemicellulose is responsible for the biodegradation, moisture
absorption, and thermal degradation of the fibre as it shows least resistance,
whereas lignin is thermally stable but is responsible for the UV degradation.31
The percentage composition of each of these components varies for different
fibres. Generally, the fibres contain 60–80% cellulose, 5–20% lignin, and up to
20% moisture (Table 6.3).31
126 Chapter 6
Figure 6.4 Structure of a bio-fibre.30
Table 6.3 Mean chemical composition of some vegetable fibres in (% of dry

mass).32
Flax Hemp Jute Abaca Coir Sisal
Cellulose 71.2 78.3 71.5 70.2 35.6 73.1
Hemicelluloses 18.5 5.4 13.3 21.7 15.4 13.3
Pectin 2.0 2.5 0.2 0.6 5.1 0.9
Lignin 2.2 2.9 13.1 5.6 32.7 11.0
Extractives 4.3 – 1.2 1.6 3.0 1.3
Fat and wax 1.6 – 0.6 0.2 – 0.3
6.4 Vegetable Fibres: Advantages and Disadvantages

Interest in vegetable fibres is growing owing to their better properties. They
possess many advantages and disadvantages compared to synthetic fibres like
glass or carbon fibres. At this time, for example, glass fibre-reinforced plastics
have been proven to meet the structural and durability demands of automobile
interior and exterior parts. However, glass-reinforced plastics exhibit short-
comings, such as their relatively high fibre density (approximately 40% higher
than vegetable fibres), machining difficulties, poor recycling properties, and the
potential health hazards posed by particulates.3 The lignocellulosic fibres,
besides their environmental advantage over synthetic ones, offer the capacity to
buckle rather than break during processing and fabrication. In addition, cel-
lulose possesses a flattened oval cross-section that enhances stress transfer by
presenting an effectively higher aspect ratio.8 The advantages and dis-
advantages are detailed next, for each property or characteristic of interest.
6.4.1 Production
Vegetable fibres need less energy in their production, resulting in lower pollu-
tant emissions. Glass fibre production requires 5–10 times more non-renewable
energy than vegetable fibre production.33
However, vegetable fibres cannot be made to grow anywhere in the world,
whereas a plant for glass or carbon fibre production can be installed nearly
anywhere, so the cost of transportation can play a significant role in the overall
cost of production, and care needs to be taken in calculating the cost of pro-
duction of each type of fibre. This difficulty can also lead to a higher envir-
onmental impact. It was shown that, for a buggy bonnet LCA (life cycle
assessment) in Brazil, in the jute fibre-reinforced composite production phase,
about 52% of the environmental impact is caused by transport fuel
consumption.34
6.4.2 Environment/Health
The most interesting aspect about vegetable fibres is their positive environmental
impact. Vegetable fibres are a renewable resource, with production requiring little
energy. They are carbon dioxide neutral, i.e. they do not return excess carbon
dioxide into the atmosphere when they are composted or combusted;8 they also
provide carbon dioxide sequestration and are biodegradable.28 They also present
safer handling and working conditions compared to synthetic reinforcements.8
It has been shown that despite the higher energy consumption to treat the jute
fibres for a buggy bonnet production, their lighter weight characteristic ensures
better environmental performance compared to glass fibre.35 The use phase of
vehicles is the most pollutant phase. For similar recycling efficiency, Silva5 has
shown that the energy required to reach their respective efficiency is about 76%
higher for glass composites than for jute composites.

The growing interest in lignocellulosic fibres is mainly due to their economical
production, with few requirements for equipment and low specific weight
compared to glass fibre composites (Table 6.4). For example, using bast fibres
in the automotive industry leads to weight savings of between 10 and 30% and
corresponding cost savings.8 It has been shown that vegetable fibres perform
better than the glass fibre composites for all fibre architectures and better than
aluminum or steel.9 A practical translation of this parameter is that a panel
with the same weight and surface area can be more than 10% stiffer in vegetable
fibre composites compared to glass.
Dent resistance is also a major factor when selecting a material for auto-
mobile enclosures. It is proportional to the square of the strength of the
material and to the fourth power of the panel thickness and inversely pro-
portional to the panel bending stiffness. Panels made of vegetable fibre com-
posites, being inherently lighter, will lead to an increase in dent resistance when
compared to glass fibre composite panels with the same weight.
One of the disadvantages is the high moisture absorption of vegetable fibres,
leading to swelling and the presence of voids at the interface, which results in
poor mechanical properties and reduces dimensional stability of composites.8,37
It was shown in jute fibre/unsaturated polyester composites that, owing to
128 Chapter 6
5,11,36
Table 6.4 Mechanical properties of fibres.
Specific Specific
Young’s elastic Tensile tensile Elongation
Density, modulus, modulus, strength, strength, at break
Fibre r (g cm–3) E (GPa) E/r s (GPa) s/r (%)
Cotton 1.51 12 7.95 0.4 0.26 3–10
Flax 1.4 60–80 42.86–57.14 0.8–1.5 0.57–1.07 1.2–1.6
Hemp 1.48 70 47.30 0.55–0.9 0.37–0.61 1.6
Jute 1.45 10–32 6.89–22.07 0.45–0.55 0.31–0.38 1.1–1.5
Abaca 1.5 31.1–33.7 20.73–22.47 0.43–0.81 0.29–0.54 2.9
Coir 1.33 4–6 3.01–4.51 0.14–0.15 0.11 15–40
Sisal 1.45 26–32 17.93–22.07 0.58–0.61 0.40–0.42 3–7
E-glass 2.6 73 28.07 1.8–2.7 0.69–1.04 2.5
water absorption in a weathering test, the flexural modulus can decrease by

about 30%.38 Another drawback of vegetable fibres is the quality variation
(even between individual plants in the same cultivation), depending on growth
conditions, processing, and other reasons, leading to non-uniformity and var-
iation of the dimensions and of their mechanical properties.8,39 Therefore,
quality control must be very rigorous for accurate results. Vegetable fibres have
problems at the interfacial adhesion between the fibre and polymer matrix,
which determines the composite physical properties, owing to the polar and
hydrophilic nature of lignocellulosic fibres and the non-polar characteristics of
most organic polymers. Because of this, it is usually necessary to make com-
patible or couple the blend.3,6,37
6.4.4 Physical and Chemical Properties

Vegetable fibres possess high electrical resistance. They have a hollow cellular
structure, providing excellent sound absorption efficiency.40 Vegetable fibres
are more shatter resistant and have better energy management characteristics
than glass fibre in their respective composites.2 Vegetable fibres in the manu-
facture of fire resistant upholstery have already been produced, where non-
woven flax (LinFR) is used to act as a fire barrier that reduces the vulnerability
of filling material to the development and spread of fire.8
An important drawback of vegetable fibres is their low microbial resistance
and susceptibility to rotting, which is a restriction to the successful exploitation
of bio-fibres for durable composites. These properties pose serious problems
during shipping, storage, and composite processing.8
6.4.5 Processing
Vegetable fibres are non-abrasive to mixing and molding equipment, which can
contribute to a significant cost reduction.8 In automotive parts, compared to
glass fibre composites the vegetable fibre composites reduce the mass of the
component, lowering the energy needed for production by 80%.2,5
The major disadvantage of vegetable fibre reinforced composites is their
inherent polar and hydrophilic nature of the lignocellulosic fibres and the non-
polar characteristics of most thermoplastics, resulting in compounding diffi-
culties and leading to non-uniform dispersion of fibres within the matrix, which
impairs the efficiency of the composite. Another problem is that the processing
temperature of composites is restricted to 200 1C, as vegetable fibres undergo
degradation at higher temperatures.31 The result of prolonged high-tempera-
ture exposure may be discoloration, volatile release, poor interfacial adhesion,
or embrittlement of cellulose components.28,30
It is quite clear that the advantages outweigh the disadvantages and most of
the shortcomings have remedial measures in the form of chemical treatment.8
However, several major technical considerations must be addressed before the
engineering, scientific, and commercial communities gain the confidence to
enable wide-scale acceptance of vegetable fibres, particularly in exterior parts
where a class A surface finish is required. Challenges include the homo-
genization of the fibre’s properties and a full understanding of the degree of
polymerization and crystallization, adhesion between the fibre and matrix,
moisture repellence, and flame retardant properties.3 Care has to be taken so as
not to erode the environmental advantage of vegetable fibres with chemical
treatments.
6.5 Applications in the Automotive Industry

Regardless of the need for low variability in mechanical properties,41 the use of
vegetable fibre reinforced composites has extended to almost all fields. In the
past decades, vegetable fibres composites with thermoplastic and thermoset
matrices have been embraced by car manufacturers and suppliers for door
panels, seat backs, headliners, package trays, dashboards, and interior
parts.3,5,8 The following are some examples of principal car component appli-
cations for various vegetable fibre types:3,8,16,42
Flax, sisal and hemp are processed into door cladding, seatback linings
and floor panels (Figure 6.5). Flax fibres are also used in car disk brakes to
replace asbestos fibres.
Coconut fibre is used to make seat bottoms, back cushions, and head
restraints.
Cotton is used to provide sound proofing; it was embedded in phenolic
resin and used in the body of the East German Trabant car, the first
production car manufactured with natural fibres.
Abaca is used in underfloor body panels.
Wood fibres are used in a large number of applications in decks, docks,
window frames, and molded panel components.
Kenaf is used in door inner panels (Figure 6.6).
130 Chapter 6
Figure 6.5 The underbody of a DaimlerChrysler A-class compression molded flax/

polypropylene.3
(a)
(b)
Figure 6.6 (a) A kenaf fibre mat; (b) a door inner panel comprising 50% kenaf mat
and 50% polypropylene.3
Recently, three-layer particleboards were produced from a mixture of sun-

flower stalks and poplar wood at certain ratios utilizing urea/formaldehyde
(UF) adhesives. Results show that all the panels provide properties required by
the normal standards for general purpose-use particleboards.8
All the major car manufacturers now use bio-based composites in various
applications. Some of these applications are presented next:2,3,7,8,40,43,44
DaimlerChrysler is the biggest proponent, with up to 50 components in its

European vehicles being produced from bio-based materials. They have
been exploring the idea of replacing glass fibres with natural fibres
in automotive components since 1991. A subsidiary of the company,
Mercedes-Benz, pioneered this concept with the ‘‘Beleem project’’ based in
Sao Paolo, Brazil. In this case, coconut fibres were used in the commercial
vehicles over a nine-year period. This initiative translates into new jobs in
coconut production, allowing the local population to improve their life
conditions. Mercedes also used jute-based door panels in its E-class
vehicles in 1996. In September 2000, DaimlerChrysler began using vege-
table fibres for their vehicle production. Johnson Controls has started
production of door-trim panels from vegetable fibre and polypropylene
for DaimlerChrysler. DaimlerChrysler has now increased its research and
development in flax-reinforced polyester composites for exterior applica-
tions. In Figures 6.7 and 6.8 are presented some examples of the appli-
cation of natural fibres in Mercedes cars.
In 2000 Audi launched the A2 midrange car in which door trim panels
were made of polyurethane reinforced with mixed flax/sisal needle felt.
Honda embarked on using vegetable fibre materials, such as wood-fibre
parts in the floor area of the Pilot sport utility vehicle (SUV), a decision
that was driven by engineering considerations as well as corporate
philosophy.
BMW Group incorporates a considerable amount of renewable raw
materials into its vehicles, including 10 000 tonnes of vegetable fibres in
2004.
General Motors kenaf and flax mixture has gone into package trays and
door panel inserts for Saturn L300s and European-market Opel Vectras,
while wood fibre is being used in seatbacks for the Cadillac Ville and in the
cargo area floor of the GMC Envoy and Chevrolet TrailBlazer.
Ford mounts Goodyear tires that are made with corn on its fuel-sipping
Fiestas in Europe. Goodyear has found its corn-infused tires have lower
rolling resistance than traditional tires, providing better fuel economy. The
sliding door inserts for the Ford Freestar are made with wood fibre. Ford
made the automotive industry’s first application of wheat straw-reinforced
plastic for the third-row storage bins of the 2010 Ford Flex. In its Start
Concept, sisal fibre is used to form the interior panels. Figure 6.9 present a
front-end grill opening reinforcement used on Ford cross members.
Toyota has shown interest in using kenaf to make Lexus package shelves,
and incorporated it into the body structure of Toyota’s i-foot and i-unit
concept vehicle. Over the course of the last five years, Toyota has eval-
uated numerous materials made from renewable resources to assess their
performance, appearance, safety, and mass production capability. They
have introduced environmentally preferable parts in a number of their
132 Chapter 6
vehicles, and currently they are investigating new materials for fabrics and
carpets as well as additional applications of PP/PLA-based and natural
fibre-based materials in North American vehicles.
An overview of some automotive usage of vegetable fibres is presented in

Table 6.5.
The recent reports are pointing out the importance of vegetable fibres and
bio-composites in the automotive industry, where many of their advantages,
like the above, are presented. The vegetable fibres incorporation in automotive
components can also play an important role towards legislations accomplish-
ment. Using materials, like vegetable fibres, into automotive parts, that can be
simple buried at the end of their life, represent an aid for accomplishment of
legislations, like the end of life vehicle (ELV) directive in Europe, which states
(a)
(b)
Figure 6.7 (a) Under floor protection trim of Mercedes A-class made from abaca
(Musa textilis) fibre reinforced composites. (b) Newest Mercedes S-class
automotive components made from different vegetable fibre reinforced
composites.8
Figure 6.8 Mercedes E-class components. By using flax/sisal thermoset in the door
panels, a 20% weight saving was achieved.40
Figure 6.9 A front-end grill opening reinforcement for the Ford cross member.3
that by 2015, that vehicles must be constructed of 95% partially decomposable

or recyclable materials. Therefore, the end of life process becomes easier, with
lower costs and less energy involved. These aspects will definitely lead to an
increased use of vegetable fibres.
134 Chapter 6
45
Table 6.5 Vegetable fibres usage in the automotive industry.
Automotive
company Vehicle Part
Audi A2, A3, A4, A6, A8, Avant, Seat back, side and back door panels,
Roadstar, Coupe boot lining, hat rack, spare tire lining
BMW 3, 5, and 7 series and others Door panels, headliner panel, boot
lining, seat back
Daimler A, C, E, S class Door panels, windshield/dashboard,
Benz business table, pillar cover panel
Fiat Punto, Brava, Marea, Alfa –
Romeo 146 and 156
Ford Mondeo CD 162 and Focus Door panels, B-pillar, boot liner
Mitsubishi Space star, Colt Door panels, instrumental panels
Opel Astra, Vectra and Zafira Headliner panel, door panels, pillar
cover panel, instrument panel
Peugeot New model 406 –
Renault Clio –
Rover Rover 2000 and others Insulation, rear storage shelf/panel
Volkswagen Golf A4, Passat Variant, Door panel, seat back, boot lid finish
and Bora panel, boot liner
Volvo C70 and V70 –
6.6 Future Perspectives

Currently, there is on-going global research into the use of vegetable fibre
composites, and automakers are producing prototypes that provide a hint into
the future of manufacturing:3,46,47
The U.S. Agricultural Research Service has been developing industrial and
commercial uses for a wide variety of agricultural products, including
waste items, and groups such as the Soybean Checkoff and the National
Corn Growers Association that focus on researching and promoting new
markets for member’s crops are supporting research efforts into new
applications for their feed sources.
Tier 1 suppliers are actively involved in producing prototype parts. Thus,
Visteon has developed a system for making flax-based instrument panels;
Composites Products has developed a process to produce door panels
from flax; Findlay Industries, which makes the cargo area floors for GM
and Honda SUVs and the package shelves for Saturn and Opel, also
manufactures headliners for Mack Trucks that are made with a hemp,
flax, kenaf, and sisal mixture; and soy-resin body panels have been
developed that are currently used on John Deere Tractors.
As can be seen in Table 6.6, the consumption of vegetable composite is

expected to increase greatly by 2020, including in the automotive industry.
By maintaining the optimization of the vegetable fibre cloth production for
composites and improving the adhesion between fibre and resin, it is clear that
the performance of vegetable fibre composites will be improved even further.9
Table 6.6 Estimated consumption evolution from 2010 to 2020.12

Estimated quantities Estimated quantities
Vegetable composites in the EU, 2010 in the EU, 2020
Compression molding 40 000 t 120 000 t
Flax, hemp, jute, kenaf, sisal, abaca,
coir (495% automotive, 5% cases
and others)
Cotton fibre (automotive, mainly trucks) 100 000 t 100 000 t
Wood fibre (mainly automotive) 50 000 t 150 000 t
Extrusion and injection molding 120 000 t 120 000 t
Wood/plastic composite (construction,
furniture, automotive, consumer
goods)
Flax, hemp, jute, kenaf, sisal, cork 5000 t 5000 t
(construction, furniture, automotive,
consumer goods)
Vegetable composites in total 315 000 t 830 000 t
Composites (glass, carbon, and natural 2.4 Mt 3,0 Mt
fibres reinforced plastics) in total
Vegetal-based share ca. 13% ca. 28%
In the end, the path will be in the direction of an important question: will
a 100% green composite for automotive components be possible? By a 100%
green composite, we mean a composite where both fibres and matrix are from
renewable sources. Therefore, to complete the set, a bio-based thermoset or
thermoplastic is needed.
Bio-resin is a term used to describe a resin or a resin formulation derived
from a biological source.48 A bio-resin can be a thermoplastic or a thermoset
biodegradable plastic.39 The most widely applied renewable resources include
plant oils, polysaccharides (mainly cellulose and starch), and proteins.39,49
In the last century, the cultivation of oilseed crops, primarily for food use, led
to the investigation of vegetable oils as precursors for resin systems.48 Examples
of vegetable oils used are from rapeseed, soybean, castor, pine, etc.39,50
Another example of a bio-resin is poly(lactic acid) (PLA), which is a bio-
based thermoplastic synthesized from lactic acid monomer. The lactic acid
monomer is produced via fermentation from corn starch. Over recent years,
owing to its good mechanical properties, much attention has been given to
potential applications for PLA as a replacement for petroleum-based
products.51
The major barriers for widespread acceptance of biopolymers, as substitutes
for traditional non-biodegradable polymers, are performance limitations and
the high cost. For example, PLA is sensitive to hydrolytic degradation under
melt processing conditions in the presence of small amounts of moisture; thus,
the hygroscopic nature of the natural fibres has a negative effect on the adhe-
sion mechanism as well as the biodegrability of bio-based composites.52
136 Chapter 6
Figure 6.10 Bio-based electric scooter, developed by NPSP Composieten and Qwic,
where the entire metal frame has been replaced by a natural fibre-
reinforced monocoque structure.9
Figure 6.11 Homogeneous components in industrial tests with hemp/PTP fibres,

showing a high surface quality.53
The high cost of biopolymers compared to traditional plastics is not due to the
raw material costs for biopolymer synthesis; rather, it is mainly attributed to
the low volume of production. Another challenge for the development of
biodegradable polymers lies in the fact that such biopolymers should be stable
during storage or usage and then degrade once disposed of after their intended
lifetime.2 With the rapid advancements in fully bio-based polymers that may be
processed with vegetable fibres, it is only a matter of time before fully bio-based
composites are prevalent within the automotive industry.3 Some examples can
be seen in Figures 6.10 and 6.11, and the components that were produced
illustrate clearly that lightweight and high surface quality construction can be
achieved by the use of natural composites.53
6.7 Conclusions
It was seen that the use of vegetable fibres in the automotive sector is real, and
the tendency is to increase their adoption. This result is mainly due to even more
stringent environmental legislations that result from the depletion and uncer-
tainty of petroleum sources, and because natural fibres have better environ-
mental performance than other traditional ones such as fibreglass. They are
from renewable sources and deliver the same performance for a lower weight.
Research on natural fibre composites will continue. Increased awareness of
environmental issues will only foster the tendency to push the boundaries even
further. The major automakers have already started using components pro-
duced by vegetal fibres in their products, and new developments are in progress.
In the direction of more sustainability, 100% ‘‘green’’ materials are being
developed. Composite materials that are 100% bio-based are being researched,
where a bio-fibre and a biopolymer are used. However, the performance
limitations and the high cost are major drawbacks to the widespread use of
biopolymers. Despite this, good results are already achieved, such as a fully bio-
based composite with high surface quality and excellent burning characteristics.
It can be said that ‘‘plant-based cars are the wave of the future’’.
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CHAPTER 7
Water Vapour Sorption of

Natural Fibres
C. A. S. HILL
Forest Products Research Institute, Joint Research Institute for Civil and
Environmental Engineering, Edinburgh Napier University, Merchiston
Campus, Colinton Road, Edinburgh, EH10 5DT, UK
7.1 Introduction
The mechanical properties of natural fibres are strongly influenced by their
moisture content and it is important to understand how and why water can
affect their behaviour. Although the water sorption properties of natural fibres
are dependent upon the presence of hydroxyl groups within the cell wall
polymers, this is not the only factor determining this behaviour. A critical
consideration is the cell wall composition and in particular the structure and
composition of the interfibrillar matrix of the cell wall. The cell wall of natural
fibres can be viewed as a composite structure, with the reinforcing elements
comprising the crystalline cellulose of the microfibrils embedded within a
matrix that is variously composed of hemicelluloses, lignin, pectin and other
substances. Water molecules sorbed within the cell wall do not penetrate the
microfibrils, but are solely located within the cell wall matrix. Water acts as a
plasticizer for the cell wall matrix macromolecules, which consequently affects
the mechanical behaviour of the fibre. Above a threshold moisture content,
natural fibres become susceptible to microbiological degradation. The presence
of water in natural fibres can also lead to deleterious properties arising from the

140
Water Vapour Sorption of Natural Fibres 141
presence of defects in the composite formed during processing. Shrinking and
swelling of fibres in response to changes in cell wall moisture content can result
in the decoupling of fibre reinforcement from the enveloping matrix, leading to
loss of performance. However, the hygroscopic properties of natural fibres can
be exploited in textile or insulation applications in the built environment. It is
clear that water sorption must be understood and if necessary controlled. In
this chapter the water sorption behaviour of natural fibres is discussed and
some methods for reducing the hygroscopic behaviour are evaluated.
7.2 The Sorption Isotherm

When a natural fibre is exposed to a constant relative humidity (RH) at a fixed
temperature, it will gradually attain a stable moisture content (MC). This
occurs when an equilibrium condition is realised and for this reason it is
referred to as the equilibrium moisture content (EMC). The equilibrium state is
dynamic and this point is reached when the rate at which water molecules are
entering the cell wall is equal to the rate at which they exit the cell wall.
Determining the EMC at a constant temperature over a range of RH values
produces a sorption isotherm. The sorption isotherm is characterized by having
a sigmoidal form and is classified as being International Union of Pure and
Applied Chemistry (IUPAC) Type 2.1 Apart from the sigmoidal shape, the
sorption isotherm of natural fibres also displays the property of hysteresis,
where the EMC values attained during adsorption are lower than those asso-
ciated with desorption at a given RH, as shown in Figure 7.1.
The sorption isotherm therefore consists of two boundary curves (adsorption
and desorption) and a scanning curve where desorption is initiated at a specific
RH and the curve passes through the region bounded by these two curves.2
Figure 7.1 Example isotherm showing (a) first desorption from saturation, (b) sub-
sequent desorption boundary curve, (c) adsorption boundary curve and
(d) scanning curve.
142 Chapter 7
In principle, every point within the space delineated by the boundary curves is
accessible by starting a scanning curve at an appropriate point on the adsorption
or desorption boundary curves. These properties have been known for over a
century, yet there is still not a consensus as to why the sigmoidal isotherm is
observed, nor is there a definitive explanation for hysteresis. The isotherm is
generally reproducible, but this property can change if the fibre is subjected to
prolonged drying at temperatures in excess of 100 1C, which can lead to degra-
dation of the cell wall polymers and also irreversible hydrogen bond formation in
the matrix polymeric network. It is also quite commonly observed that when a
never-dried fibre is taken through the initial desorption stage, the path of this
first desorption branch of the isotherm is not reproduced in any subsequent
desorption experiments. Two explanations can be posited for this behaviour.
Firstly, it cannot be guaranteed that all of the water present in the fibre when
drying down from a saturated state is actually located within the cell wall; sec-
ondly, it is possible that there is some irreversible hydrogen bond formation
(hornification) occurring in the initial drying process, irrespective of whether this
takes place at elevated or ambient temperatures. In practice, the determination of
sorption isotherms is undertaken over a RH range from zero to 95%, since it
becomes very difficult to make accurate measurements much above this upper
RH limit. There is also an increasing likelihood of a contribution from capillary
condensation to the sorption process at RH values exceeding 90%.3
There is considerable variation in the sorption isotherms determined for
different fibre types.3 The reasons for this are complex, but are undoubtedly
related to the cell wall composition and microfibril angle (MFA), although the
importance of hydroxyl group availability is not clear. The importance of
the cell wall composition is related to the mechanical response of the cell wall
to the presence of sorbed moisture and the MFA will also influence the
mechanical properties of the cell wall. For example, a high MFA will allow for
considerable linear extension of the fibre when moisture is adsorbed onto the
internal surface. Figure 7.2 shows comparative isotherms for flax and coir,
where it can be seen that the fibre with the highest lignin content (coir) also
exhibits the highest levels of water sorption. Coir also exhibits a high MFA (of
the order of 401) compared with flax (around 31).
Given that lignin has a much lower hydroxyl (OH) to carbon ratio compared
with polysaccharides, this result at first appears to be counter-intuitive. How-
ever, although the presence of hydroxyl groups undoubtedly is a major factor in
making the fibres hygroscopic, the ability and extent of swelling of the cell wall
to accommodate the sorbed water molecules is probably the most significant
factor in determining the EMC over most of the hygroscopic range. Thus a
mechanical model for describing water sorption would appear to be more
appropriate.
Although sorption isotherms are not determined at RH values much in
excess of 95%, it is quite common practice to extrapolate the adsorption iso-
therm line through the data points and project onto the EMC axis at 100% in
order to determine a ‘‘fibre saturation point’’ (FSP).4 The term ‘‘fibre satura-
tion point’’ is used to represent the moisture content of the fibre in a theoretical
Figure 7.2 Isotherms for coir and flax.
condition where the cell wall moisture content is at a maximum, but there is no
water present in the macrovoids of the fibre. In the case of wood, it is estab-
lished that the FSP is a point where many of the physical properties (e.g.
swelling, modulus of elasticity, modulus of rupture, impact toughness) no
longer change and this is often found to be in the region of 30% MC for wood
and many plant fibres.2,5 However, in practice it becomes very difficult to
measure the changes in properties in the region 30–40% MC and there is some
dispute as to where the ‘‘true’’ FSP may lie. Methods which produce total
saturation of the cell wall with water indicate that the FSP may lie nearer to
40% MC.3 The use of the projection of adsorption isotherms to evaluate a FSP
has been criticized on a number of grounds.6 One objection relates to the fact
that the sorption isotherm is changing rapidly at this point and small errors in
curve fitting lead to large errors in the projected FSP (p-FSP) value. It is also
not possible to determine isotherms with any degree of accuracy much above
95% RH and it is then necessary to resort to methods such as tension plates. In
addition, it has been found that there is some variation in the isotherm at the
higher end of the hygroscopic range below 95% RH even with identical fibre
samples, as is shown for flax in Figure 7.3.
Despite these criticisms, the projection method is still commonly used to
report FSP. This has been done for a range of natural fibres, where it can be
seen that there is considerable variability in the p-FSP (Table 7.1).3 It is
immediately obvious from these data that cotton has a much lower p-FSP
compared to all the other fibres in this table.
Although caution must be exercised when interpreting the p-FSP values
for reasons noted above, it is possibly instructive to compare the results of
Table 7.1 with the polymeric composition of these fibres (Table 7.2).3
Cotton, with a very high cellulose content, has the lowest p-FSP, which could
perhaps indicate that it is the lack of accessible OH groups that is determining
the EMC. It is striking that coir, which has the highest lignin content of the
144 Chapter 7
Figure 7.3 Reproducibility of the sorption isotherm of flax.
Table 7.1 The projected fibre saturation point of

different natural fibres.
Fibre p-FSP (%)
Jute 27.0
Coir 34.0
Flax 25.6
Hemp 30.4
Cotton 17.0
Table 7.2 The polymeric composition (wt%) of the natural fibres listed in
Table 7.1, where ‘‘polysaccharide’’ refers to all cell wall poly-
saccharides other than cellulose.
Fibre Cellulose Polysaccharide Lignin
Jute 72 14 13
Coir 43 5 45
Flax 81 16 3
Hemp 74 19 4
Cotton 94 6 0
fibres in this study, nonetheless also has the highest p-FSP. This suggests that
invoking accessible OH content as the only variable affecting EMC is unlikely
to be correct.
7.3 Water Sorption Models

A large number of models exist describing the sorption process; these can be
classified as localized models, which can be broadly split into layering or cluster
models,7,8 and dissolution models.9 In localized models, the water molecules are
considered to bind to specific sites in the polymeric matrix. The layering model
requires the initial formation of monolayers on the cell wall internal surface,
with the subsequent creation of multiple layers as the sorption process pro-
ceeds. Indeed, there is a significant component of the literature which reports
on the monolayer content, often determined by the application of Brunauer–
Emmett–Teller (BET) isotherm analysis.10 The application of such a model can
only strictly be made in the case of an inert substrate, not a swelling cell wall
(elastic gel) which will present an evolving surface as the adsorption process
takes place. Nonetheless, BET analyses are commonly made with water vapour
sorption isotherms and BET surface areas are reported from these analyses. A
modification to the BET model has led to the development of the Guggenheim–
Anderson–de Boer model and its variants.11 In dissolution models a macro-
scopic approach is adopted where there is mixing between macromolecules and
water molecules; examples include the Hailwood and Horrobin (HH) model,
which was originally developed to explain the sorption isotherm of cotton,12
and the Flory–Huggins model.13 The HH model is based upon consideration of
the thermodynamics of the system where a sorbent is in equilibrium with the
atmospheric moisture. The analysis considers the equilibrium existing between
the solid dry polymer of the sorbent, hydrated polymer and ‘‘dissolved’’ water.
The hydrated polymer component is water that is strongly bound to the
sorption sites of the polymeric constituents of the substrate, whereas the dis-
solved water is water that is not so strongly bound but is still located within
the cell wall nanopores. These two types of water can be loosely interpreted
as monolayer and polylayer water analogously to the BET model, although the
concept of a monolayer existing with the complex, dynamic geometry of the cell
wall internal surface is unrealistic. An HH analysis of the sorption isotherms of
jute is shown in Figure 7.4. The sum of the two types of cell wall water produces
a sigmoidal curve that very closely fits the data, except at the upper end of the
Figure 7.4 Hailwood–Horrobin model of the sorption isotherm showing the con-
tributions from dissolved water and water of hydration.
146 Chapter 7
hygroscopic range where it is possible that there is an additional contribution
from capillary condensation. It is important to note that the HH model cannot
be used to analyze the desorption isotherm curve, since this is composed of a
boundary and a scanning curve.
7.4 Hysteresis
A number of explanations have been given in the plant fibre and wood science
literature to explain hysteresis, but these have been unsatisfactory for a variety
of reasons.3 Early explanations relied upon the assumption that hysteresis
merely represented the failure to achieve ‘‘true’’ equilibrium in the experiment
and that if a sufficient amount of time was allowed then the equilibrium state
would be achieved. It has been shown in numerous studies that this is not the
case and that hysteresis is a true phenomenon and not an experimental artefact.
Other explanations have considered the nature of the adsorption process
occurring on the surfaces of the cell wall nanopores, resulting in a stepwise
filling which continues until the nanopore is filled with liquid water. Desorption
then occurs from the surface meniscus of the water in the nanopore, giving rise
to hysteresis. However, the concept of the existence of a meniscus in liquid
water in these cell wall nanopores, which are of the order of 2–4 nm in diameter,
is problematical, making such an explanation highly suspect. It has been argued
that hysteresis arises due to the formation of irreversible hydrogen bonds
during the first desorption cycle, which can certainly be used to explain why the
first compared to subsequent desorption boundary curves do not follow the
same path, but cannot explain why adsorption and desorption boundary curves
follow different paths on subsequent sorption cycles. Another model relies
upon consideration of the geometry of the nanopores in the cell wall, where the
diameter of the throat of such a capillary is smaller than the interior, akin to an
ink bottle. Other explanations invoke differences in contact angle between the
adsorption and desorption processes. During the wetting cycle the contact
angle of the water with the internal surface of the cell wall is different compared
with that for desorption, where the water is in contact with an already wet
surface. The problem with such an explanation is that the use of concepts
related to liquid water in such small capillaries is unlikely to have any physical
meaning; furthermore, during the adsorption process, the water is entering
from a vapour state and not as a liquid. It is however possible that such an
explanation may have some application in the upper part of the hygroscopic
range (above 70% RH), where capillary condensation becomes more impor-
tant. It has been suggested that hysteresis is caused by the presence of per-
manent gases in the cell wall, leading to incomplete wetting, but it has been
shown that hysteresis is still observed even if sorption isotherms are determined
in a vacuum. With this model under conditions of adsorption the sorption sites
are thought to be partially masked by the presence of gas molecules. It has also
been argued that the presence of permanent gases in such samples only con-
tributes towards hysteresis if such materials are non-swelling.3
When water vapour enters the cell wall, it causes swelling of the material.
This is because the water is occupying space between the microfibrils, thereby
forcing them apart (water cannot penetrate the microfibrils). This process of
expansion is resisted primarily by the interfibrillar matrix polymers; as water is
removed from the cell wall, the matrix collapses to its previous configuration,
i.e. the nanopores are not permanent but transient. The cell wall of plant fibres
and many other natural materials (e.g. wood) can be considered to be a swelling
gel.14 The sorption behaviour of natural fibres has been reported upon recently,
where a model for sorption hysteresis was presented based upon the micro-
mechanical behaviour of the cell wall matrix during the adsorption or deso-
rption process.3 This model was originally developed to describe sorption in
glass polymers15 and subsequently applied to sorption hysteresis effects with
humic soils.16 Essentially, the process of adsorption results in the creation and
expansion of nanopores within the cell wall matrix, whereas desorption leads to
the collapse of these voids in the cell wall matrix. This process is inelastic on the
time scale of molecular diffusion and as a consequence the adsorption and
desorption processes take place in a material that is in different states. This
phenomenon is observed in glassy polymers below the glass transition tem-
perature (Tg).3,17 The glassy state is defined as being one in which the molecular
scale nanopores are embedded in a matrix that is unable to fully relax to a
thermodynamic equilibrium state due to the stiffness of the matrix macro-
molecules. It is therefore logical to consider that there is a link between the
matrix mechanical properties and the hysteresis effect. It has been found that
the extent of hysteresis (i.e. the area bounded by the adsorption and desorption
curves) is related to the cell wall composition of fibres and that fibres with
higher lignin contents display greater hysteresis, which is consistent with the
model.3 This is illustrated in Figure 7.5, showing a comparison of the hysteresis
between the adsorption and desorption isotherms (obtained by subtracting the
Figure 7.5 Differences in hysteresis (calculated by subtracting adsorption from des-

orption moisture contents).
148 Chapter 7
Figure 7.6 Effect of temperature on the isotherm of flax.
EMC of adsorption from the EMC of desorption) for flax, hemp, coir and jute.
The fibres with a higher lignin content display a greater amount of hysteresis
between the adsorption and desorption boundary curves.
According to the model, the presence of a cross-linked matrix polymer (such
as lignin) would be expected to result in a higher level of hysteresis, but the
model also predicts that as the isotherm temperature increases, the extent of
hysteresis should decrease and become zero at the Tg of the matrix. Figure 7.6
shows the effect of temperature upon the sorption isotherm of flax. As
predicted by the model, the hysteresis decreases as the isotherm temperature
is raised. This property has been previously noted with Sitka spruce18 and
cotton.19
7.5 Water Adsorption and Cell Wall Swelling

As noted in the previous section, the adsorption of moisture into the cell wall of
a fibre leads to swelling of the material. The diffusion of water molecules into
the cell wall matrix that surrounds the microfibrils requires the breaking of
hydrogen bonding networks, with the creation of nanopores.3,20 It is thought
that the initial water molecules entering the cell wall are relatively tightly bound
to the primary sorption sites (OH groups), but as the adsorption process
continues, the water molecules that diffuse into the cell wall nanopores are less
constrained in their translational freedom.21 As the cell wall swells, this creates
elastic strain in the matrix. The sorbed water molecules therefore perform work
upon the cell wall and during the reverse process this strain energy is released as
water molecules leave the cell wall. However, the cell wall deformation process
is not perfectly elastic since hysteresis occurs in the sorption isotherm. This is
further discussed in Section 7.7.
7.6 Sorption and Heat of Wetting
Water molecules in the vapour state have considerably more translational
energy than they do in the liquid state and this energy is released as heat when
fibres adsorb water. This is the same as the latent heat of vaporization of water,
but there is an additional amount of heat (heat of wetting) that is released
because of the additional constraint imposed on the water molecules by being
confined within the cell wall nanopores. There are two ways of measuring the
heat of wetting: either directly by using calorimetry, or indirectly by applying
the Clausius–Clapeyron equation.3 Although widely adopted within the sci-
entific literature, the application of a thermodynamic analysis to a non-rever-
sible system is unsound and indeed this is one criticism of the HH model, which
incidentally is unable to explain hysteresis. Below the glass transition tem-
perature of the matrix, the sorption phenomenon is not path independent.
7.7 Kinetics of Water Sorption

Many models describing the kinetics of water sorption processes within natural
fibres have unconvincingly invoked Fick’s Law.22 One of the confounding
factors when attempting to model sorption kinetic behaviour has been the
difficulty of obtaining sufficiently accurate data sets, a requirement for reliable
curve-fitting. However, with the commercial development of dynamic vapour
sorption (DVS) equipment over the past decade, it has now become a routine
matter to obtain extremely accurate experimental kinetic data. Using a DVS
apparatus, it was clearly demonstrated by Kohler et al.23 that the sorption
kinetic behaviour with flax fibres is actually very accurately described by what
is termed a parallel exponential kinetics (PEK) model, which has the mathe-
matical form shown in eqn (7.1):
MC ¼ MC0 þ MC1 ½1 expðt=t1 Þ þ MC2 ½1 expðt=t2 Þ ð7:1Þ

where MC is the moisture content at time t of exposure of the sample to a
constant RH and MC0 is the moisture content of the sample at time zero. The
sorption kinetic curve is composed of two exponential terms which represent a
fast and slow process having characteristic times of t1 and t2, respectively. The
terms MC1 and MC2 are the moisture contents at infinite time associated with
the fast and slow processes, respectively. The PEK model is not just applicable
to flax fibres,23,24 but also to a wide range of plant fibre types,25 regenerated
cellulose fibres26–28 and microcrystalline cellulose,29 as well as unmodified and
modified wood,30,31 but has also been found to be applicable to the drying
behaviour of foodstuffs and the swelling properties of cereal grains.32–34
Although not yet proven, it is likely that the PEK model can be applied to all
swelling gel materials, with the proviso that the sample size (or more correctly
the volume to surface ratio) is small, although what the limiting ratio is has not
been determined. An example sorption kinetics curve is shown in Figure 7.7 for
flax and jute fibre, with the PEK fit to the data given and the fast and slow
adsorption curves also reproduced.
150 Chapter 7
Figure 7.7 Sorption kinetic curves of flax and jute deconvoluted into fast and slow
components according to the parallel exponential kinetics model.
12
10 COTTON FLAX
adsorption fast
8 adsorption slow
MC (%)
desorption fast
6 desorption slow
0
0 20 40 60 80 100 0 20 40 60 80 100
RH (%) RH (%)
Figure 7.8 Pseudo-isotherms for the fast and slow processes shown for cotton and
flax.
It is not known what these two processes represent, but they have been
previously attributed to the existence of ‘‘fast’’ and ‘‘slow’’ sorption sites in
the cell wall.23 The fast sites have been attributed to the sorption of water
molecules associated with the external surface and amorphous regions of the
fibre and the slow sites to indirect sorption on the inner surface and crystal-
lites.26 By cumulatively adding the MC1 or MC2 values for adsorption or the
desorption cycles, it is possible to construct pseudo-isotherms associated with
the fast and slow processes and consequently determine what the contribution
of the two processes is to the hysteresis effect. This is shown for cotton and
flax in Figure 7.8.
It has been noted by workers that the adsorption pseudo-isotherms resemble
the isotherms attributable to monolayer (Langmuir) and poly-layer adsorp-
tion.23 However, it has been shown that such an interpretation of the fast and
slow processes to ‘‘fast’’ and ‘‘slow’’ sorption sites is not appropriate.35,36
An alternative model for interpreting the sorption kinetics is presented in
Section 7.8. It is important to note when comparing different fibre samples that
the sample weights should be closely comparable ( 10%) and that the same
RH steps should be used in the isotherms. This is because the time constants are
dependent not only upon morphological and compositional differences between
fibres, but also upon sample weight and RH step size.
7.8 Water Sorption and Mechanical Behaviour

It is well known that the modulus of elasticity and strength of wood37 and
paper38 decrease as the moisture content is raised up to the fibre saturation
point, whereas the toughness of the material increases with the MC. However,
reliable data for the influence of moisture on isolated plant fibre mechanical
properties are not abundant. The reasons for this can be attributed to:
The inherently high variation in fibre properties, plus the effect of fibre
damage on mechanical properties.39
Variation due to test fibre length or testing speed.40
Difficulties in obtaining accurate fibre cross-sections (the reason why the
textile industry uses linear density), plus the change in cross-sectional area
as the MC of the fibre varies.
Small sample sets.
Poorly reported data (RH and temperature not reported, or not
controlled).
Conditioning regime not reported (conditioning down from a high to a
lower specific RH will not give the same EMC as when conditioning from
a low to a higher specific RH).
Not testing using the same RH and temperature at which the fibres were
originally conditioned.
It is well known that plant fibres exhibit a decrease in modulus as the RH of

the testing environment increases, although the change in ultimate tensile
strength does not display such a simple relationship.2 Fibres containing a high
content of amorphous polysaccharide material would be expected to exhibit
reductions in tensile strength and modulus from consideration of the properties
of the isolated material.41 Such systems can be considered to act as viscoelastic
gels, which will display creep behaviour when subjected to long-term loading
effects. The mechanical behaviour of such systems can be modelled as a com-
bination of elastic and plastic deformations, as is well known when examining
the mechanical behaviour of wood.42 Since water is able to plasticize the cell
wall matrix substances of plant fibres, it also follows that the static and
dynamic mechanical properties of the fibres will be modified accordingly.2 The
literature appears to be silent on the effect of duration of load upon natural
fibre reinforced composites and the effect of variations in atmospheric RH
upon this behaviour. However, there has been some work remarking upon the
non-linear stress–strain behaviour of unidirectional flax reinforced composites,
152 Chapter 7
Figure 7.9 Two Kelvin–Voigt elements in series.
behaviour attributed to the annealing of microcompressive deformations in the

fibres.43 This is clearly an area where research needs to be directed if natural
fibre reinforced composites are to be used in structural applications.44
While the mechanical behaviour of plant fibres is modified by the presence of
cell wall moisture, it is also probable that the sorption behaviour is determined by
the mechanical properties. It has recently been argued that the sorption kinetics
of these gel-like materials is rate limited by the rate of swelling of the sorbent and
that the kinetic process is in fact controlled by the matrix polymeric relaxation
processes.45,46 Based on this premise, the PEK model has been further interpreted
in terms of two Kelvin–Voigt (KV) viscoelastic elements acting in series (Figure
7.9) with E1 and E2 being the moduli associated with the fast and slow processes,
respectively, and Z1 and Z2 being the equivalent matrix viscosities.
The two elements represent the fast and slow kinetics processes. The kinetic
response of a KV element when subjected to an instantaneous stress increase
(s0) is as shown in eqn (7.2):47
e ¼ ðs0 =EÞ½1 expð t=jÞ ð7:2Þ
where e is the strain at time t, E is the elastic modulus and j is a time constant
which is defined as the ratio Z/E, with Z the viscosity. In the case of a plant
fibre subjected to a change in RH, there is a change in the swelling pressure
(P, equivalent to s0) exerted within the cell wall when the atmospheric water
vapour pressure is raised from an initial value pi to a final value pf given by
eqn (7.3):48
Q
¼ ðr=MÞRT lnðpi =pf Þ ð7:3Þ
where r is the density and M is the molecular weight of water, R is the gas
constant and T is the isotherm temperature in kelvin. In the model described
herein, the strain of the system is assumed to be equivalent to the volume
change of the cell wall as a result of water vapour adsorption or desorption.
This volume change is further assumed to be linearly related to the change in
the mass fraction of the water present in the cell wall.
The adsorbed water vapour molecules exert a pressure within the cell wall
leading to dimensional change, which is equivalent to the extension of the
40 40
E1ads ads
1
30 E1des 30 des
1
E2ads
(TPa s)
ads
E (GPa)
2
E2des
20 20 des
2
10 10
0 0
0 20 40 60 80 100 0 20 40 60 80 100
(a) RH (%) (b) RH (%)
Figure 7.10 Moduli and viscosities for the cell wall calculated from the PEK para-
meters and applying the KV model.
spring in the KV model. This expansion/extension represents work and results

in an increase in the free energy of the system. Expansion will continue until the
free energy of the system is equal to the free energy of the water vapour
molecules in the atmosphere. The spring modulus (E1, E2) therefore defines the
water content of the system at infinite time (MC1, MC2), as shown in eqn (7.4).
The fact that there is a relationship between the EMC of a fibre and modulus
was established over 50 years ago.49
MC1 ¼ s0 =E1 ; MC2 ¼ s0 =E2 ð7:4Þ

The rate at which water molecules are adsorbed or desorbed by the system is
a function of the viscosity of the dashpot in the model. This viscosity is in turn
related to the micro-Brownian motion of the cell wall macromolecular network.
The more rapidly the matrix is able to deform, the faster the rate of water
ingress or egress into or out of the cell wall. The rate of local deformation is
related to the energy barrier associated with the local relaxation process and
whether there is sufficient free volume to allow the relaxation process to take
place. In glassy solids below the glass transition temperature (Tg) there is
insufficient free volume to allow a local relaxation to take place without the
cooperative motion of adjacent relaxors (a relaxor is defined as the smallest
molecular segment of relaxation in each polymeric unit). This gives rise to the
concept of cooperative domains within the matrix.50–52 As the glass transition
temperature is approached, the domain size decreases until Tg is reached. At
this point the domain contains only one relaxor and there is sufficient free
volume to allow for relaxation without the cooperation of neighbours. When
this occurs, matrix relaxation becomes instantaneous and the behaviour is
perfectly elastic. Hysteresis is no longer observed.
Results from the KV interpretation of the PEK data for x and y are given in
Figure 7.10. By applying the sorption kinetic parameters to the KV model
representing the viscoelastic response of the cell wall matrix, it is possible to
derive values for the matrix modulus and viscosity. This has been done for the
matrix modulus associated with the fast process (E1) and the slow process (E2)
154 Chapter 7
and these data are presented in Figure 7.10. Values for the fast and slow moduli
are of the order of 10–30 GPa at the low cell wall moisture contents associated
with the bottom end of the hygroscopic range. These moduli decrease to very
low values at the higher RH range, as would be predicted for a matrix that is
being plasticized by the presence of sorbed water.
These modulus values at low RH are comparable with tensile modulus
properties reported in the literature for these fibres.53 Given that the expansion
and contraction of the cell wall takes place within the intermicrofibrillar matrix,
it seems reasonable to assume that the modulus values obtained are dominated
by those associated with the matrix polymers, e.g. pectin, hemicellulose and
lignin. Unfortunately, there is little in the way of reliable direct measurement of
the mechanical properties of these substances in the scientific literature.
Salmén41 has discussed the subject of cell wall polymer elastic constants at
some length. Although the elastic modulus values for isolated lignin and
hemicellulose have been determined, there is no reason to suppose that this is a
true measure of their behaviour within the cell wall. Assuming that these
reported values are representative, it is apparent that the modulus values cal-
culated using the KV model in this study for the dry cell wall are considerably
higher at low cell wall MC. This indicates that the moduli also have a con-
tribution from the deformation of the cell wall microfibrils. Although Salmén
quotes axial microfibrillar modulus values of 134 GPa, the off-axis modulus is
given as 27.2 GPa, but is not affected by moisture.41 Thus the dry modulus
values calculated using the KV model in this study appear to be of the order
expected, given contributions both from the matrix and microfibrils, but the
microfibrillar contribution decreases at higher cell wall MC. The explanation
for this lies in consideration of the geometry of the microfibrils within the cell
wall. For example, it has been shown with jute fibres that water adsorption and
desorption result in a rotation of the fibre about the long axis as well as an
increase in cell wall volume.54 This indicates that the distortion of the micro-
fibrils occurring as a result of cell wall volume changes does not involve a
simple off-axis bending motion. It may be that verification of the applicability
of the KV interpretation of the sorption kinetics could be obtained by nano-
indentation methods. The author of this chapter is not aware of any such
studies with plant fibres, but there have been investigations of the properties of
the wood cell wall. Nano-indentation methods have given values for the
modulus of the order of 13–21 GPa for the cell wall of spruce (moisture content
not stated),55–57 comparable with the low MC modulus values reported herein.
Differences in behaviour between fibres are also found when the matrix
viscosities are examined for the fast process (Z1) and the slow process (Z2). For
the viscosities associated with the slow process, the Z2 values are higher for
desorption, although there are differences in the behaviour between the fibres
studied. Since the fast and slow processes have not been assigned to specific
physical properties, it is not at this stage possible to comment upon the sig-
nificance of such differences. What is clear is that the viscoelastic interpretation
does seem to provide an insight into the sorption process and that there are
important variations in behaviour that require further study.
It has previously been argued that an appropriate model describing sorption
hysteresis in plant fibres and wood is related to the matrix response during
adsorption/desorption.3 As discussed earlier, the model describes the creation
of nanopores in the matrix during the adsorption step and the collapse of these
nanopores during desorption.15–17 However, below the Tg of the matrix, the
matrix is unable to respond instantaneously to the ingress or egress of water
molecules because of the structural rigidity of the matrix. This results in an
increased nanopore volume in the matrix during the desorption compared to
the adsorption process, leading to an increased affinity for the water molecules.
The timescale of the nanopore formation/annihilation processes are molecular
(i.e. of the order of 1010 s). By invoking the concept of cooperative relaxation
in polymer systems below Tg, Matsuoka50 was able to show that relaxation
times of the order of minutes can be observed as the size of the cooperative
domain increases (i.e. as the temperature decreases). This suggests that there
should be a link between the hysteresis effect and the relaxation times observed
in the sorption process.
7.9 Methods to Reduce Water Sorption

As noted previously, thermal modification of plant fibres by prolonged heating
at temperatures above 160 1C reduces the hygroscopicity of fibres, but is also
responsible for degrading the mechanical properties.58 A commercial thermal
modification process (Duralin) was developed for flax fibres over a decade ago,
but this is no longer in commercial production. Thermal modification removes
hygroscopic polysaccharide material in the cell wall and can also result in cross-
linking promoted by the formation of furfuryl alcohol produced from degraded
pentoses.59 Other methods of reducing water vapour sorption require the
chemical modification of the cell wall macromolecules, or impregnation of the
cell wall with monomers, with subsequent polymerization in order to prevent
leaching in service. A comparison of the water vapour sorption isotherms of
flax and Duralin is shown in Figure 7.11.
Acetylation has been studied extensively as a means of improving the
dimensional stabilization of wood and its potential for protecting natural fibres
from degradation and the effects of water. The method used in commercial
wood modification is to react the substrate with acetic anhydride at tempera-
tures in excess of 120 1C. This leads to esterification of the cell wall polymeric
OH groups and generates acetic acid as a by-product which must then be
removed from the modified material (Figure 7.12).
Apart from removal of the OH groups by reaction, the bonded acetyl groups
also occupy space within the cell wall, resulting in a permanent swelling
(bulking). Removal of the OH groups by substitution with the less polar acetyl
groups affects wettability, but the reduction in EMC is due to the bulking
effect.60 In studies of wood modified with a variety of anhydride reagents, it was
shown that the reduction in EMC was related only to the weight gain due to
bonded reagent, but was not related to the number of OH groups substituted.60
156 Chapter 7
Figure 7.11 Sorption isotherms of flax and Duralin.
Figure 7.12 The acetylation reaction with acetic anhydride.
It has been shown that significant improvements in performance are found

when acetylated fibres are subjected to biological degradation.61 The improved
resistance to biodegradation is again found to be independent of the extent of
OH substitution and appears to be due to the reduction of the cell wall MC
caused by the filling of internal space in the cell wall due to the presence of
bonded reagent.62 It is known that fungal attack of the wood cell wall requires
the MC to be in excess of 20% and it appears that acetylation is able to provide
protection by ensuring that the cell wall MC is below this threshold.
It has also been found that improvements in resistance to the degradation of
composites reinforced with silane-treated fibres can be achieved.63 Silanes have
long been used for sizing of glass fibres and they can also be used as a means of
promoting adhesion between fibre and matrix, depending on the chemistry of
the silane. There has been considerable interest in the use of organo-functio-
nalized alkoxysilanes (Figure 7.13) in order to improve the bonding between
natural fibres and matrix in composite materials,64 but under certain conditions
it is also possible to modify the interior of the cell wall and hence reduce water
sorption. However, it is necessary to carefully control the modification condi-
tions in order to optimize cell wall penetration by the reagent. Polymerization
of the silane can occur once the alkoxy groups are removed by hydrolysis, but
(a)
(b)
Figure 7.13 (a) Reaction of an alkoxy organosilane with water and subsequent
polymerization, and (b) reaction of an organosilane with the cell wall
polymers and subsequent hydrolysis.
such reactions are undesirable, at least until the silane monomer has penetrated
the cell wall. Premature oligomerization of the silanes prevents penetration of
the cell wall and restricts reactions to the surface of the fibre.65
7.10 Conclusions
The water sorption behaviour of natural fibres is an important property that
influences their mechanical behaviour. It is proposed herein that there is a link
between the mechanical properties of the fibre cell wall, the sorption kinetics
and the property of hysteresis. The sorption kinetic behaviour can be very
accurately described using the parallel exponential kinetics model, whereas
Fickian behaviour is not observed. It is proposed that the sorption kinetics
behaviour is described by consideration of the viscoelastic relaxation properties
of the cell wall polymers. In addition, the cell wall polymer relaxation beha-
viour can be invoked as a means of explaining hysteresis. This then suggests
158 Chapter 7
a link between mechanical properties and water sorption behaviour that should
prove a promising line of research for the future.
Acknowledgement
The author acknowledges the support of the Scottish Funding Council for
funding of the Joint Research Institute in Civil and Environmental Engineer-
ing, which is part of the Edinburgh Research Partnership in Engineering and
Mathematics.
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Academic Press, London, 2nd edn., 1982.
2. W. E. Morton and J. W. S. Hearle, Physical Properties of Textile Fibres,
CRC Press, Boca Raton, 4th edn., 2008.
3. C. A. S. Hill, A. Norton and G. Newman, J. Appl. Polym. Sci., 2009,
112, 1524.
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CHAPTER 8
Environmentally Friendly
Coupling Agents for Natural
Fibre Composites
R. CHOLLAKUP,*a W. SMITTHIPONGa AND
P. SUWANRUJIb
a
Kasetsart University, Kasetsart Agricultural and Agro-Industrial Product
Improvement Institute (KAPI), 50 Ngam Wong Wan Rd., Chatuchak,
10900, Bangkok, Thailand; b Department of Chemistry, Faculty of Science,
Kasetsart University, 50 Ngam Wong Wan Rd., Chatuchak, 10900,
Bangkok, Thailand
8.1 Introduction
In recent years, natural source materials have gained attention because they
are renewable, widely distributed, locally accessible, environmentally recycl-
able, conveniently available in many forms and biodegradable. In particular,
natural fibres, i.e. kenaf, flax, jute, hemp, sisal, pineapple, silk, etc, are
abundant worldwide as biomaterial sources.1–6 These fibres are based on either
cellulose or protein. Cellulose is used to reinforce the structure of plant cells,
whereas silk, a protein fibre produced from silkworms, is used to build the
cocoon for protecting the silkworm from predators during its metamorphosis
into a moth. Thus, these natural fibres have received much interest as possible
reinforcing materials for plastic composites in many applications such as

161
162 Chapter 8
7,8
automotive and package products. The advantages of these biomaterial
sources, compared to traditional reinforcing materials such as glass, aramid
and carbon fibres, are their low cost, low density, toughness, acceptable spe-
cific strength properties, reduced tool wear, reduced dermal and respiratory
irritation, good thermal properties, enhanced energy recovery and, especially,
biodegradability.9–11
Fibre-reinforced composite materials are part of the general class of engi-
neering materials called composite materials. A useful definition of composite
materials is to state that composite materials are characterized by being
multiphase in which the phase distribution and geometry have been deliber-
ately tailored to optimize one or more properties.12,13 This is clearly an
appropriate definition for fibre-reinforced composites for which there is one
phase, called the matrix, reinforced by material in the form of a fibre. Gen-
erally, the fibre is stronger than the matrix. When a composite is forced by
external action, this force is transferred from the matrix to the fibre, which
increases its strength compared to the matrix without the fibre. The size and
shape of fibres play an important role in the efficiency of reinforcement. In
addition to a wide choice of materials, there is the added factor of the man-
ufacturing route to consider, because a valued feature of composite materials
is the ability to manufacture the article at the same time as the material itself is
being processed.
The use of natural fibres for the reinforcement of composites has received
increasing attention both by the academic sector and industry. Currently, many
types of natural fibres14,15 have been investigated for use in plastics, including
flax, hemp, jute straw, wood, rice husk, wheat, barley, oats, rye, cane (sugar
and bamboo), grass, reed, kenaf, ramie, oil palm empty fruit bunch, sisal, coir,
pennywort, kapok, paper mulberry, raphia, banana fibre, pineapple leaf fibre
and papyrus. Thermoplastics reinforced with special wood fillers are enjoying
rapid growth due to their many advantages: lightweight, reasonable strength
and stiffness. Some plant proteins are interesting renewable materials because
of their thermoplastic properties. Wheat gluten,16 once plasticized, is unique
among cereals and other plant proteins in its ability to form a cohesive blend
with viscoelastic properties. For these reasons, wheat gluten has been utilized to
process edible or biodegradable films and packaging materials. Hemp17 is a
bast lignocellulosic fibre, which comes from the plant Cannabis sativa and has
been used as a reinforcement in biodegradable composites.
Generally, the properties of composites depend on the matrix and the fibre.
The development of new composite materials based on natural fibres has been
driven by interesting new applications. The role of the matrix in a fibre-rein-
forced composite is to transfer stress between the fibres, to provide a barrier
against an adverse environment and to protect the surface of the fibres from
mechanical abrasion. The matrix plays a major role in the tensile load-carrying
capacity of a composite structure. The binding agent or matrix in the composite
is of critical importance. Here, we summarize three types of matrix: (i) non-
biodegradable synthetic polymers in Table 8.1; (ii) biodegradable synthetic
polymers in Table 8.2; and (iii) biopolymers in Table 8.3.
Environmentally Friendly Coupling Agents for Natural Fibre Composites 163
Table 8.1 Examples of natural fibre composite materials based on non-bio-
degradable synthetic polymers.
Matrix Natural fibre Improvement properties Ref.
Epoxy Sisal Tensile strength 18
Phenol/formaldehyde Palm Tensile strength and 19,20
(PF) modulus, dynamic
mechanical properties
Polycarbonate Pineapple Mechanical properties 21
Polyethylene (PE) Coir and palm Young’s modulus 22
Pineapple Dynamic storage and loss 23–25
modulus, tensile strength
Sisal Tensile strength, electrical 26–28
properties, degradation
Polypropylene Palm Tensile strength, flexural 30,31
strength and modulus,
water absorption
Pineapple Tensile strength 25,29,32
Sisal Tensile strength, dynamic 33,34
mechanical properties,
thermal degradation
Polystyrene (PS) Palm Modulus 35
Sisal Thermal stability 36
Poly(vinyl chloride) Palm Flexural modulus 37
(PVC) Rice straw Tensile strength 38
8.2 Interface between the Natural Fibre

and the Matrix
Multicomponent materials and, in particular, reinforced materials have known
a huge industrial development in the last few decades. There has also been an
increasing interest in the scientific community to analyze and understand the
physical and chemical nature of the interface between the reinforcing entity and
the matrix. More precisely, good final mechanical performance or use prop-
erties of the resulting reinforced composites depend significantly on the quality
of the interface that is formed between both solids. In other words, the adhe-
sion established at this interface becomes one of the most important parameters
in controlling the interfacial behaviour, and consequently, the mechanical
properties of reinforced materials. Therefore, it is understandable that a better
knowledge of the adhesion phenomena is required for practical applications of
multicomponent materials. Consequently, the study of adhesion uses various
concepts, depending on different special fields of expertise. This variety
of approaches is emphasized by the fact that many theories of adhesion have
been proposed which together are both complementary and contradictory,
i.e. mechanical interlocking, electronic or electrostatic theory, theory of
weak boundary layers, thermodynamic or adsorption model (also referring to
wettability), diffusion or interdiffusion theory, and finally, chemical bonding
theory.69–72
164 Chapter 8
Table 8.2 Examples of natural fibre composite materials based on biode-
gradable synthetic polymers.
Poly(butylene Bamboo Tensile properties, water resis- 39
succinate) tance, interfacial adhesion
(PBS) and enzyme biodegradability
Poly(butylene Wheat straw Mechanical properties 40
adipate-co-
terephthalate
Poly- Cellulose fibre Electrical conductivity 41
caprolactone
(PCL)
Polyester Jute Tensile strength 7
amide (PEA) Sisal Mechanical properties 42
Polyester Banana Dynamic mechanical 43
properties
Poly(hydroxy- Flax Bending modulus and dynamic 44
butyrate) mechanical properties
(PHB)
Poly(hydroxy- Pineapple Tensile and flexural strength 45,46
butyrate vale- and modulus
rate) (PHBV)
Poly(lactic Bamboo Impact strength, heat resis- 41,42,47
acid) (PLA) tance, water resistance, inter-
facial adhesion and enzyme
biodegradability
Cotton with Tensile strength and modulus 48
lignin
Flax Impact, tensile strength, 49,50
Young’s modulus
Hemp Tensile strength, modulus and 51,52
flexural strength and ease of
thermal degradation
Kenaf Mechanical and thermal 53–55
properties
Reed Mechanical properties 56
Silk Reinforcement for PLA for 57
tissue engineering
Wood Fluxural modulus 58
Poly(vinyl Baggase Thermal stability 58
alcohol) Lignocellulosic Enhance PVA degradation 59
(PVA) fibre
Unsaturated Hemp Impact and tensile strength 60
polyester
(UPE)
However, the thermodynamic model of adhesion, generally attributed to

Sharpe and Schonhorn,73 is certainly the most widely used approach in adhe-
sion science at present. In this theory, it is considered that adhesion between
two solids (more generally, between an adhesive and a substrate) is due to
Table 8.3 Examples of natural fibre composite materials based on
biopolymers.
Thermoplastic starch Flax Tensile strength and modulus 61
Sisal Mechanical properties 62
Cellulosic fibres Tensile strength and modulus 63
from Eucalyptus
urograndis pulp
Leafwood cellulose, Mechanical properties (tensile 64
paperpulp fibre tests), thermo-mechanical
properties (DMTA) and
thermal degradation
Soy protein isolate Ramie Mechanical properties 65
(SPI)
Cross-linked soy flour Flax Tensile and flexural properties 67
(CSF)
Corn gluten meal Wood Mechanical properties 66
(CGM)
Modified acrylated Flax and hemp Tensile strength, flexural 68
expoxidized soy oil strength and modulus
(AESO)
interatomic and intermolecular forces established at the interface, provided that

an intimate contact is achieved.74 The most common interfacial forces result
from van der Waals and Lewis acid–base interactions. The magnitude of these
forces can generally be related to fundamental thermodynamic quantities, such
as surface-free energies of both entities in contact. Generally, the formation of
an assembly is obtained through a liquid–solid contact step. Therefore, criteria
for good adhesion become, essentially, criteria for good wetting, although this
is a necessary but not a sufficient condition. It is clear that when an intimate
contact has been established at an interface, other adhesion phenomena can
therefore take place, in particular the creation of interfacial chemical bonds, the
molecular reorganization and reorientation near the interface, the interdiffu-
sion of molecular or macromolecular chains across this interface in the case of
contact involving polymers, etc. Each of these phenomena is able to enhance
the level of adhesion.75–82
In fact, each of these adhesion theories is validated by some measurements,
depending on the nature of the materials and the formation conditions of the
adhesive junction. Although each theory is supposed to explain the adhesion
in different ways, it in fact contributes to this complex phenomenon. Cur-
rently, six main mechanisms involving chemical or physical forces are con-
sidered. Their actions at the interface between two materials, the same or
different, are shown in Figure 8.1. These mechanisms are only validated at
different scales by the distance between the two materials: the atomic scale for
the chemical bonds, a few angstroms for intermolecular bonds, a scale not
known (but reasonably within the range of micrometers) in the case of
mechanical bonding.
166 Chapter 8
(a) (b)
(c) (d)
(e) (f)
(g)
Figure 8.1 Model of adhesion theory: (a) mechanical bonding; (b) electrostatic
bonding; (c) interfacial layers of low cohesion; (d) adsorption; (e) diffusion
theory; (f) chemical bonding theory; and (g) chemical bonding of coupling
agents between the fibre surface and the polymer matrix.
8.3 Types of Coupling Agents

Coupling agents are compounds that function at the interface of natural fibres
and the polymer matrix to form a bridge of chemical and/or physical bonds
between the two phases. Generally, coupling agents possess two functions: the
first is to react with the fibre surface, i.e. at the polar hydroxyl (OH) group, and
the other is to react with functional groups of the polymer matrix. In this
heterogeneous system, coupling agents couple the two phases together via
various interactions, such as covalent bonding, hydrogen bonding, acid–base
interaction and chain entanglement, depending on the types of polymers and
fibres and the functional groups of the coupling agents. The incorporation of
coupling agents in fibre-reinforced polymer composites facilitates the formation
of interfacial bonding between the fibres and the matrix, leading to the
improvement of mechanical properties from effective stress transfer and bond
distribution across the interface. In most cases, selection of a coupling agent for
fibre-reinforced polymer composite is based on the combination of strength
and toughness at the fibre/matrix interface. The most common coupling agents
used for natural fibre-reinforced polymer composites are silane, isocyanate and
maleated coupling agents.10,83–85
8.3.1 Conventional Coupling Agents

8.3.1.1 Silanes
Silane coupling agents are effectively used in natural fibre/polymer composites
because of their bifunctional structures that can enhance the interfacial adhe-
sion between the fibre reinforcement and the polymer matrix. The general
chemical structure of a silane is R–SiX3, where R represents an organofunc-
tional group which can react with the polymer matrix and X are groups that
can hydrolyze to form silanol groups in an aqueous solution.10,83,84 Examples
of silane coupling agents are shown in Table 8.4. Castellano et al. reported that
silane itself does not react with the hydroxyl groups of cellulose fibres even at
high temperatures, because of the low acidity of cellulose hydroxyl groups.86
Table 8.4 Silane coupling agents for natural fibre/polymer composites

(R–SiX3).
Function-
Name R X ality
Hexadecyltrimethoxy-silane –(CH2)15CH3 –OCH3 Alkyl
(HDS)
Dichlorodiethylsilane (DCS) –(CH2)2CH3 –Cl2 Alkyl
g-(Aminopropyl) –(CH2)3NH2 –OCH2CH3 Amino
triethoxy-silane (APS)
(Cyanoethyl) –(CH2)2CN –OCH3 Cyano
trimethoxy-silane (CES)
g-(Mercaptopropyl)- –(CH2)3SH –OCH3 Mercapto
trimethoxysilane (MPS) (thiol)
g-(Methacryloxypropyl)- O –OCH3 Methacryl
trimethoxysilane (MCS)
(CH2)3 O C C CH2
CH3
Vinyltri(2-methoxyethoxy)- –CH¼CH2 –OCH2CH2OCH3 Vinyl
silane (VTS)
168 Chapter 8
However, the hydrolyzable X groups can form the more reactive silanols in the
presence of moisture (eqn 8.1), which then react with hydroxyl groups of
natural fibre macromolecules:87,88
RSiX3 þ 3H2 O ! RSiðOHÞ3 þ 3XCl

ð8:1Þ
silane silanol
Owing to the high affinity for each other, silanol groups can also undergo
self-condensation to form oligomers via –Si–O–Si bonds. Consequently, not
only the silanol monomers but also the oligomers adsorb to the hydroxyl
groups of the fibre by the formation of hydrogen bonds. Under thermal con-
ditions, the hydrogen bonds formed between the adsorbed silanols and the
hydroxyl groups of the fibre can be converted into covalent bonds of –Si–O–C
linkages,89–91 as shown in Figure 8.2.
The R groups at the other end of the silane can be a large number of func-
tional groups, as shown in Table 8.4. These functional groups play an impor-
tant role in the interaction between the silane and the polymer matrix used,
depending on their functionality’s reactivity towards the matrix. The non-
reactive groups of silane, i.e. alkyl groups, would be expected to increase the
compatibility with the hydrocarbon backbone of the polymer matrix, such as
polyethylene (PE) and polypropylene (PP). On the other hand, reactive groups
such as vinyl, methacryl and amino groups can form a chemical bond with a
OH
HO Si R OH
H
O H OH O O Si R
Siloxane Adsorption H OH
OH OH H OH OH
O H O
HO Si O Si R O Si O Si R
R OH H OH OH
Fibre Oligomer
(Grafting)
O
O Si R
O O
O Si O Si R
O
R
Figure 8.2 Mechanism of the interaction between silanes and cellulose fibres.
88,90,92–98
polymer matrix containing reactive functional groups. Therefore, it is
important to find a proper match between the silane functionality and the
target matrix to improve the interfacial adhesion. The interaction between the
silane group and the polymer matrix may be van der Waals forces95 or by a
grafting reaction.93 Covalent bonds can also form between aminosilanes and
epoxy resins.97,98
8.3.1.2 Isocyanates
Isocyanates function as useful coupling agents by the formation of covalent
bonds between the hydroxyl groups of cellulose fibres and the isocyanate
(–N¼C¼O) functional groups of the coupling agents.85,88,99,100 The most
common isocyanate coupling agents are shown in Table 8.5. The urethane or
carbamate linkage between cellulose fibres and isocyanate coupling agents can
be formed as shown in eqn (8.2). The formation of carbonyl groups on the fibre
surface has been confirmed by Fourier transform infrared spectroscopy
(FTIR).88
R N C O + HO Cell R N C O Cell ð8:2Þ
Since natural fibres are highly hygroscopic with a great tendency to absorb
water, isocyanate coupling agents are susceptible to hydrolysis. In the presence of
even traces of moisture, isocyanates prefer the reaction with water to cellulosic
Table 8.5 Isocyanate coupling agents for natural fibre/polymer composites.

Name Structure
Methylene
diisocyanate (MDIC)
O C N N C O
Polymethylene
polyphenyl isocyanate
(PMPPIC)
O C N N C O
n
Toluene CH3
2,4-diisocyanate (TDIC)
N C O
O C N
Hexamethylene O C N
diisocyanate (HMDIC) N C O
170 Chapter 8
100
hydrogen groups, producing polyurea, biuret and isocyanate dimers. There-
fore, the use of blocked isocyanates has been reported to prevent the hydrolysis of
isocyanates prior to reaction with the fibre and also its thermal dissociation. The
enhancement of mechanical properties and water resistance of wheat starch/PLA
blends was reported as a result of the addition of methylene diisocyanate
(MDIC).101,102 George et al. claimed that the very high interfacial interaction
between pineapple-leaf fibre (PALF) and a PE matrix was because of the addition
of polymethylene polyphenyl isocyanate (PMPPIC). An effective water uptake
reduction of the composite was due to the improvement of interfacial bonding
from the coupling effect of PMPPIC.88 The maximum retention in strength of
PMPPIC-treated PALF/linear density polyethylene (LDPE) composites at
higher temperatures was also reported.32 The isocyanate groups in PMPPIC are
highly reactive with the hydroxyl groups of fibres and a urethane linkage can be
formed between them. The long-chain molecules in PMPPIC interact with the PE
matrix through a van der Waals interaction. The addition of toluene 2,4-diiso-
cyanate (TDIC) to short sisal fibre-reinforced polyester (PS) composites showed a
reduction of water uptake and an improvement of dimensional stability and
tensile properties. The p-electrons of benzene rings in TDIC, similar to PMPPIC,
lead to stronger interactions with the PS matrix.103 By the reaction of cellulosic
hydroxyl groups and isocyanate groups of HMDIC, the polarity and hydro-
philicity of cellulose fibres decreased and consequently improved the compat-
ibility of the cellulose fibres and PP matrix.100 Alkenyl isocyanates have also been
used in fibre/polymer composites, such as 3-isopropenyl-a,a 0 -dimethylbenzyl
isocyanate (TMIC) and 2-isocyanoethyl methacrylate (IEMC). The isocyanate
groups ensure the coupling reaction with the cellulosic hydroxyl groups while an
alkenyl moiety subsequently copolymerizes with the matrix monomers such as
styrene and methyl methacrylate.104
8.3.1.3 Maleated Coupling Agents

Maleic anhydride (MA) is an a,b-unsaturated carbonyl compound having a
C¼C double bond on the heterocyclic ring conjugated with two carboxylate
groups. MA is extensively used to graft onto polymers, especially PE and PP, to
form maleated coupling agents such as maleated polyethylene (MAPE) and
maleated polypropylene (MAPP). The anhydride groups of the maleated
copolymer can react with the surface hydroxyl groups of the fibre reinforce-
ment through esterification or hydrogen bonding, while the other end of the
copolymer can undergo entanglement interlocking with the polymer matrix as a
result of their similar polarity.105,106 FTIR and electron spectroscopy for che-
mical analysis (ESCA) have presented evidence for chemical bridges between
the wood fibre and the polymeric matrix through esterification. The two pri-
mary covalent bonding products to link the wood fibre and polymer matrix at
the interface were succinic esters and half-esters.106
The inclusion of MAPP in wood-reinforced PP composites gives a significant
improvement in the strength properties of the composites.107,108 The flexural
properties, impact strength and tensile modulus of PALF-reinforced PP com-
posites were considerably improved in the presence of MAPP.29 The mechan-
ical properties of jute-reinforced PP composites were also reported to be
improved by the application of MAPP, owing to the chemical bonds at the
fibre/matrix interface established from MAPP coupling agents.109,110 The
improvement in the abrasive wear resistance of jute-reinforced PP composites
with the addition of MAPP was also claimed by Chand and Dwivedi.111
8.3.1.4 Dichlorotriazines
Dichlorotriazines and their derivatives contain reactive chlorines on the het-
erocyclic ring that can react with the hydroxyl groups of natural fibres through
an ether linkage. Moreover, the nitrogen in the heterocyclic ring may also link
the hydroxyl groups via hydrogen bonding. Zedorecki and Flodin confirmed
the reaction between the hydroxyl groups of cellulose fibres and triazine deri-
vatives using FTIR and X-ray photoelectron spectroscopy (XPS). They
reported that triazine compounds can improve the environmental ageing
behaviour of cellulose fibre-reinforced polyester composites.112–114 Joly et al.
used dichlorotriazine substituted with an octadecyl chain as a coupling agent in
cotton/PP composites. The chemical reaction of triazine with hydroxyl groups
only occurred at the extreme surface of the fibre, while the octadecyl chain
entangled with the PP matrix. There was no measurable effect on the water
uptake of the composites.115
8.3.2 Bio-based Coupling Agents

With the world’s increased environmental awareness, it would be good if
coupling agents used in natural fibre/polymer composites were non-toxic and
biodegradable to preserve the renewable and biodegradable character of the
natural fibre reinforcement. In order to produce completely biodegradable
composites without emitting toxic or noxious compounds, a bio-based coupling
agent is needed. Therefore, many environmentally friendly coupling agents
have been reported.
8.3.2.1 Cardanol Derivative of Toluene Diisocyanate (CTDIC)

Cardanol is the main component of cashew-nut-shell liquid obtained from the
plant Anacardium occidentale, which is plentiful in tropical countries. CTDIC
was prepared from the reaction between cardanol and toluene diisocyanate.
The resulting product contained one free isocyanate group for further reaction
with the hydroxyl groups of fibres. The formation of a covalent bond between
isocyanate groups and fibres reduced the hydrophilic nature of the cellulosic
fibre. The long-chain structure of cardanol in CTDIC helps the fibre have a
higher hydrophobic character, better compatibility and dispersibility in a
matrix. Owing to a strong interfacial bond between sisal fibres and LDPE, the
172 Chapter 8
CTDIC-treated fibre composites exhibited superior tensile strength and mod-
ulus than the untreated composites. This strong fibre/matrix adhesion in the
composites was supported by scanning electron microscopy (SEM) photo-
micrographs. The dielectric constant values of sisal/LDPE composites were
found to decrease as a result of the addition of CTDIC.27,116
8.3.2.2 Lysine Diisocyanate (LDIC)

Conventional isocyanates have limited uses as coupling agents in view of
environmental concerns because their ultimate hydrolysis products, such as
4,4 0 -methylenedianiline and 2,4-diaminotoluene, are suspected cancer agents
and produce hepatitis in humans. LDIC prepared from lysine can be used as a
coupling agent without producing toxic products during its degradation. Lysine
is one of the essential amino acids, having two amino groups and one carboxyl
group. LDIC can react with hydroxyl or carboxyl groups in poly(lactic acid)
(PLA) or poly(butylene succinate) (PBS) and form hydrolyzable urethane
bonds. Tensile properties and water resistance were improved by using LDIC in
corn starch/PBS biocomposites. The enhancement of interfacial adhesion
between corn starch and a PBS matrix was a result of the coupling effect of
LDIC, even with a very small addition (o1 wt%).117 Lee and Wang also
reported the effect of LDIC as a coupling agent on the properties of bio-
composites from PLA, PBS and bamboo fibre (BF). They concluded that the
addition of LDIC improved the tensile properties, water resistance and inter-
facial adhesion of both PLA/BF and PBS/BF composites. The crystallization
temperatures of both composites increased, whereas the enthalpy and heat of
fusion decreased with increasing the amount of LDIC. However, the compo-
sites with LDIC exhibited higher thermal degradation temperatures than those
without LDIC. Moreover, LDIC also delayed enzymatic degradation because
stronger interfacial adhesion reduced the area exposed to enzyme hydrolysis.39
In corn starch/PLA composites, isocyanate groups reacted with terminal
hydroxyl or carbonyl groups of PLA and the hydroxyl groups of corn starch.
The urethane linkage was confirmed by nuclear magnetic resonance spectro-
scopy (NMR).118
8.3.2.3 Chitin and Chitosan

Chitin and chitosan are long, linear, natural polysaccharide polymers com-
posed of N-acetylglucosamine and glucosamine, respectively. Chitosan is thus
the deacetylated form of chitin. The advantages of these two compounds are
their wide availability, non-toxicity, biocompatibility and lower cost compared
with many synthetic coupling agents. Chitin and chitosan have been used as
coupling agents for wood flour/PVC composites. With the addition of chitin
and chitosan coupling agents, the flexural strength, flexural and storage
modulus of the composites increased compared with ones without coupling
agents. The property improvement by the addition of chitin and chitosan was
attributed to strong interfacial adhesion through acid–base interactions
between the chlorine-containing PVC and the amino groups on the wood
surface treated with chitin and chitosan. The optimum amount of chitin applied
was 6.67 wt%. With up to 0.5 wt% of chitosan, a significant improvement in
composite properties was attained. This difference in the amount applied was
explained by the more reactive amino groups of chitosan over the less reactive
acetylamino groups of chitin.119
8.3.2.4 Zein
Zein is an alcohol-soluble natural protein extracted from corn. Zein is micro-
bial resistant and possesses the benefits of being renewable and biodegradable.
The high molecular weight amphiphilic molecule contains an abundance of
carboxyl, amino, phenyl and methyl groups. In 1938, zein became commercially
available and quickly found applications in coatings, films, plastics, adhesives,
inks and fibres. Zein in combination with vegetable oils and glycerine as
plasticizers was used as a waxing or glaze, in order to enhance the shelf life of
pharmaceutical tablets, nuts and candies by acting as a water and oxygen
barrier. Zein has been used as a coupling agent for flax/PLA composites. The
polar groups, i.e. carboxyl and amino groups, of the zein protein react with the
hydroxyl groups of the fibres through hydrogen bonding, while aryl and alkyl
groups react with alkyl groups of PLA through hydrophobic interaction.120,121
The improvement of the storage modulus in flax/PP composites was due to
enhanced interfacial addition from the addition of zein coupling agents.122 The
composites of kenaf fibre and PP with the addition of zein coupling agents also
possess improved mechanical and viscoelastic properties.120
8.3.2.5 Fatty Acid Derivatives

Corrales et al. reported the use of oleoyl chloride, a fatty acid derivative, as a
coupling agent for cellulose/polymer composites. Oleoyl chloride is able to
react with cellulosic hydroxyl groups by an esterification reaction to form an
ester functionality. Esterification is limited to the hydroxyl groups on the
external surface of the fibre. The long hydrocarbon chain of oleoyl chloride
induces hydrophobicity on the fibre surface and can improve the compatibility
between the treated fibre and polymer matrix.123
8.3.2.6 Shellac
Shellac, which is a natural, biodegradable, renewable resource product made by
bugs, is known to be compatible with cellulose materials like jute, wood, etc.
Recently, jute/fibre-glass reinforced sheets were prepared with shellac. Their
flexural strength values were found to be in the range of those reinforced sheets
prepared using fibre glass alone. Flame retardance and thermal resistance of the
jute/fibre-glass reinforced sheets were improved with shellac. Lamination of
174 Chapter 8
plywood and particleboard was also investigated with jute/fibre glass and
shellac, and improved mechanical properties were obtained.124 Shellac can be
used as a matrix to prepare biocomposites made of natural fibre (jute) using
biodegradable coupling agents (urea) to ensure superior mechanical
properties.125
8.3.2.7 Fungal Modifications

A fungus can selectively degrade non-cellulosic compounds, particularly lignin,
at a faster rate than they degrade wood cellulose. This reduction of non-cel-
lulosic compounds is also believed to be responsible for the observed increase in
both crystallinity and thermal stability of the fibre. In addition, fungal treat-
ment is suggested to increase the surface roughness of the fibre. Microorgan-
isms of Ophiostoma ulmi were used to modify the properties of hemp fibres for
composite reinforcement. O. ulmi treatment slightly improved the moisture
absorption characteristics of natural fibres, which should benefit the durability
of the composites. Treated hemp fibres showed improved acid–base char-
acteristics between fibre and resin, which are expected to improve the interfacial
adhesion. Good interfacial adhesion will lead to both better performance and
better durability of the composites by eliminating voids at the interface, which
act as reservoirs for the accumulation of water.126
8.3.2.8 Furfuryl Alcohol Modifications

Recently, a new selective chemical modification of the surface of lignocellulosic
natural fibres has been considered. This modification is based on the selective
oxidation of guaiacyl and syringyl units of lignin, generating ortho- and para-
quinones that are able to react by Diels–Alder reaction with furfuryl alcohol;
the latter is commercially prepared by reduction of furfural, which in turn is
obtained from agricultural residues.
A process was set for chemically modifying sugar cane bagasse and curaua
fibres. A quite specific oxidation by chlorine dioxide of syringyl and guaiacyl
phenols of the lignin polymer, creating quinones, was carried out. The latter
were reacted with furfuryl alcohol, creating a coating around the fibre more
compatible to phenolic resins. This modification favoured the fibre–matrix
interaction at the interface for sugar cane and curaua fibre-reinforced com-
posites, but caused some fibre degradation that affected the mechanical prop-
erties and decreased the mechanical resistance of the composites reinforced
with them.127 The fibre modified with furfuryl alcohol exhibited degradation of
hemicelluloses, but cellulose maintained most of its crystallinity.128 The che-
mical modification of coir fibres was also studied through oxidation with
chlorine dioxide, followed by reaction with furfuryl alcohol. This surface
modification reduced the hydrophilicity of coir fibres, leading to an improve-
ment in their moisture resistance capability. The modified fibres were more
compatible with the polymeric matrices.129
8.3.2.9 Guar Gum
Guar gum is a natural accumulated hydrocolloid stored in the endosperm of
the seeds of the guar plant. The macromolecular structure of guar lies between
a spherocolloid (like amylopectin) and a linear hydrocolloid (like cellulose).
Like most polysaccharides, guar has two or three free hydroxyl groups on the
mannose unit of the main chain or the galactose side chains. These are available
for bonding and can be utilized in bonding with the numerous hydroxyl groups
present in the jute fibre structure. The use of guar gum as a surface treating
agent for jute yarns had an increasing effect on the performance of the treated
jute/vinyl ester resin composites. However, the improvement was evident only
at an optimum concentration of 0.2%. With a further increase in guar gum
concentration (to 0.3%), the high viscosity of the solution might have caused a
thicker coating on the jute fibre surface, which resulted in an insufficient
penetration of the resin into the fibres, lowering the mechanical properties of
the composites.130
8.3.2.10 Enzyme Treatments

Some published reports have described the effect of enzymatic degradation of
non-cellulosic compounds of hemp fibres.131 Pectinases degrade pectin and
laccases modify lignin. However, waxes and other non-cellulosic compounds
can be a barrier to these enzymes. In order to achieve good results, these
enzyme treatments have been preceded by pre-treatments, such as chelator
treatment with ethylenediaminetetraacetic acid (EDTA).132 Chelating agents
are organic compounds capable of forming covalent bonds with metals through
two or more of their atoms. It has been reported that some chelators such as
EDTA can remove calcium ions from pectin in plant cell walls such as in hemp
fibres, resulting in the pectin becoming soluble in many liquids. This allows the
hemp fibres to separate from their bundles. However, EDTA persists in the
environment and, due to its strong metal chelating properties, enhances the
mobility and bioavailability of contaminant heavy metals. An alternative is
ethylenediaminetetramethylenephosphonic acid (EDTMPA), a phosphonated
analogue of EDTA. EDTMPA has a very strong interaction with all mineral
surfaces,133 so it is easily removed from technical and natural systems. Owing to
this strong adsorption, little or no remobilization of metals occurs. Therefore,
compared to EDTA, EDTMPA has less impact on the environment.
Pectinases can be broadly classified into acidic and alkaline, based on their
pH requirement for optimum enzymatic activity. Alkaline pectinases, which
come mostly from bacterial sources, are capable of degrading pectin in the
middle lamella of a fibre cell wall. They are widely used in the separation of
bundles in crops such as flax, hemp and jute to obtain fibres.134 Laccases are
one of the most important lignin degrading enzymes. It has been reported that
laccase alone cannot depolymerize lignin,131 but when 1-hydroxybenzotriazole
hydrate (HBT) is used as a mediator, a degree of delignification up to 40% has
been obtained. The mechanism of laccase–mediator systems involves oxidation
176 Chapter 8
of the mediator by laccase, followed by oxidation of lignin by the low molecular
weight oxidized mediator, which can diffuse into the structure to oxidize the
lignin molecule.
8.4 Pros and Cons of Using Bio-based

Coupling Agents
Using natural fibres as reinforcement in composites involves several challenges.
The first and the most important problem is the low fibre/matrix adhesion. The
role of the matrix in a fibre-reinforced composite is to transfer the load to the
stiff fibres through shear stresses at the interface. This process requires good
bonding between the polymeric matrix and the fibres. Poor adhesion at the
interface means that the full capabilities of the composite cannot be exploited
and leaves it vulnerable to environmental attack, thus reducing its life span.
Natural fibres are hydrophilic and polar in nature, whereas common matrices
are hydrophobic and non-polar. Natural fibres used in matrices are therefore
dimensionally unstable and display insufficient adhesion between matrix and
fibre, which results in poor composite mechanical properties. It is possible to
improve bonding between the fibre and the matrix by modifying the surface of
the fibre, or by modifying the matrix with the addition of a coupling agent or
bio-based coupling agents.
Because bio-based coupling agents are inexpensive and highly eco-friendly
materials, their use in either their raw form or some suitably modified form
could be very effective for the composite industry. A fully biodegradable green
composite from the combination of natural fibres and biodegradable resins
could be achieved by rendering bio-based coupling agents. These green com-
posites could be completely decomposed into innocuous products. Compared
with conventional coupling agents, bio-based coupling agents themselves are
considered as green chemicals because they are from renewable feedstocks,
preserve efficacy of function while reducing toxicity and do not persist in the
environment at the end of their use.1 It is known that a silane coating of glass
fibres is extensively used in the glass fibre composite industry for improved
bonding between the glass fibres and the polymer resin. However, for natural
fibres, such a universal coating material is not yet available that could be used
effectively and extensively for specific natural fibres to make them more suitable
as reinforcements in polymer composites. The use of inexpensive natural
materials, as a coupling agent or surface treating agent of natural fibres, can be
a techno-commercially viable one. In contrast, during the raw material storage
required for the industrial production of composites, bio-based coupling agents
would degrade due to standard bacterial and fungal processes. Assessing the
degradation that occurs during storage, such that it could be taken account of
in the composite processing, is recommended for future research. For example,
in the case of fungal modifications, care must be exercised in choosing the
treatment time since longer treatment times can give the enzymes secreted by a
fungus an opportunity to penetrate deep into the crystalline region of the
cellulose, which may affect the strength of the fibres. Moreover, the con-
centration of bio-based coupling agents may be optimized, depending on the
natural fibre and the nature of the matrix.
8.5 Conclusions
Natural fibres, when used as reinforcement, compete with such synthetic fibres
as glass fibre. The advantages of synthetic fibres are their good mechanical
properties, while their disadvantage is the difficulty in recycling. By use of
proper processing techniques, fibre treatments and coupling agents, several
natural fibre composites reach the mechanical properties of glass fibre com-
posites. Natural fibres are renewable, light in weight, low in cost and envir-
onmentally friendly. To enhance the interfacial adhesion between the fibre
reinforcement and the polymer matrix, different types of natural and synthetic
coupling agents have been used in natural fibre/polymer composites. Recently,
there has been increasing interest in commercialization of natural fibre com-
posites and their use, e.g. in the automobile and furniture industries. However,
poor wettability and insufficient adhesion with resins limits the use of natural
fibres in high-performance applications. In order to overcome this limitation,
bio-based coupling agents like zein, fatty acid derivatives, enzyme treatments,
etc., have been successfully used. These bio-based coupling agents are effective
and environmentally friendly methods to improve the interfacial bonding
between the natural fibre and the matrix. The future trends of natural fibre-
reinforced composites are to use bio-based coupling agents and new types of
natural fibre which could be compatible with the environment and widen the
applications of natural fibre composites, such as scaffolds for tissue engineering
applications.
Acknowledgement
We gratefully acknowledge Kasetsart University Research and Development
Institute (KURDI) and KAPI, Kasetsart University in Thailand, for their
financial support.
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CHAPTER 9
Probing Interfacial Interactions

in Natural Fibre Reinforced
Biocomposites Using Colloidal
Force Microscopy
G. RAJ,* E. BALNOIS,* C. BALEY AND Y. GROHENS
Université de Bretagne Sud (UBS-Ueb), LIMATB (Laboratoire d’Ingénierie

des MATériaux de Bretagne), rue de St Maudé, 56321 Lorient, France
*Email: [email protected]; [email protected]
9.1 Introduction
Growing environmental concerns have led recent research to focus on the
exploitation of natural fibres as reinforcements in composite materials. Natural
fibres such as flax, hemp, jute, sisal, and coir have already been used as rein-
forcements in thermoplastic composites for applications in various industries,
mainly the automotive,1 construction,2 and packaging industries. Certain
natural fibres, such as flax fibres, have demonstrated specific strength properties
close to that of glass fibres and can potentially replace them in fibre-reinforced
plastics (FRP).3 Recently, new ‘‘eco-composites’’ or biocomposites with
interesting mechanical properties have been developed from natural fibre
reinforced biodegradable polymers such as poly(lactic acid) (PLA).4 PLA has
attracted much attention, not only because of its biodegradability and

183
184 Chapter 9
biocompatibility, but also because its price has become competitive due to the
continuous increasing price of petroleum products.
Even though natural fibres present several advantages in terms of low cost,
low density, acoustics, and biodegradability properties, their efficiency as
reinforcement elements in composite materials depends on several key para-
meters. Firstly, the intrinsic strength of the fibres, which in turn is determined
by the complex hierarchical organization of several polysaccharides that con-
stitute the fibre’s internal structure;5,6 and secondly, the interfacial adhesion
between the natural fibre and the polymer matrix. The last parameter plays an
important role in efficient stress-transfer at the fibre/matrix interface region,
and a clear understanding of this adhesion phenomenon is required to develop
high-performance biocomposites suitable for structural applications.
Many attempts have been made to characterize interface adhesion in natural
fibre reinforced composite materials. In most studies, an improvement in
macroscopic material properties, e.g. tensile strength,7–9 water absorption
properties,10,11 were attributed often indirectly to an improvement in fibre/
matrix interface bonding. More recently, the adherence or ‘‘practical adhesion’’
between vegetal fibres and polymer matrix in biocomposite materials was
analysed by estimating interfacial shear stress (IFSS) values obtained from
different micromechanical tests, such as the pull-out test,12 microbonding test,13
and single fibre fragmentation tests.14 For example, Arbelaiz et al.7 used pull-
out tests on the flax/polypropylene system to investigate the efficiency of their
fibre surface treatments. The authors mentioned that the lack of homogeneity
makes the interpretation of these tests difficult and reported relatively weak
increases of IFSS values.
Nardin et al.15 proposed a semi-empirical linear relationship between IFSS
and the reversible work of adhesion established at the interface of model
composite materials involving glass or carbon fibres in thermoplastic or ther-
mosetting matrices. A remarkable feature of this relation is that, apart from the
mechanical properties of the materials involved, the IFSS (a mechanical
quantity) was completely dependent on the level of molecular interactions or
the work of adhesion (a thermodynamic quantity) at the interface. In the case
of biocomposites, a direct application of the Nardin–Schultz relation was dif-
ficult, as shown by earlier studies,13 and was attributed to the complex com-
posite structure of flax fibre. It should be noted that the work of adhesion
estimated in these studies was obtained from the surface energy values based on
the contact angle of different liquid probes on flax fibres.13 Given the rough,
heterogeneous, and porous build-up of flax fibres, the work of adhesion
obtained from the contact angle measurements lacks homogeneity, and should
be treated with caution. Complementary techniques are thus essential for a
better understanding of the complex interactions between natural fibres and
polymer matrix in a biocomposite.
Colloidal force microscopy (CFM) has emerged as a direct technique to
quantitatively estimate intermolecular forces between two approaching sur-
faces by measuring force–distance curves.16,17 The technique was, for example,
extensively used to study the interaction between cellulose surfaces in the pulp
Probing Interfacial Interactions in Natural Fibre Reinforced Biocomposites 185
18–24
and paper industries. Most of the experiments were mainly focused on the
preparation of ultra-flat cellulose surfaces and smooth, spherical, and non-
porous cellulose microbeads in order to mimic a sphere–plane interaction.
These experiments were performed in liquid or dilute electrolyte concentrations
which essentially simulate the manufacturing conditions involved in the pulp
and paper production. This approach forwarded a better understanding of the
interactions between various materials and additives in paper making.
Recently, atomic force microscopy (AFM) was found to be a valuable
complementary technique to characterize surface properties of natural fibres at
the nanoscale.25,26 It provides direct adhesion force measurements between a
standard silicon tip and the fibre25,27,28 or even directly between the polymer
matrix and cellulose.29 The quantitative interpretation of adhesion force
measurements in AFM is nonetheless limited to a precise knowledge of the
contact area between the two objects, and working with rough and hetero-
geneous fibres remains a problem. In this work, we propose to decompose and
study some of the main adhesion forces present in biocomposites. Molecularly
smooth model surfaces, which mimic the three major polysaccharides in flax
fibres, were prepared and colloidal force measurements between these surfaces
and a PLA bead, assuming sphere–plane contact geometry, were performed
and compared. Figure 9.1 represents a schematic description of the experi-
mental set-up developed that allows us to gain information on some important
adhesion mechanisms at the PLA/flax biocomposite interface.30
Force measurements were first performed under ambient conditions and
recorded under various relative humidity conditions. From the AFM pull-off
force values, the work of adhesion for the different interaction systems at the
interface region were estimated and used to verify the Nardin–Schultz rela-
tion15 for the flax/PLA biocomposite.
Cellulose RMS 4 nm
PLA bead
Xyloglucan
RMS 0.3 nm
RMS 1 nm
PGUA
RMS 0.3 nm
(a) (b)
Figure 9.1 (a) SEM image of the flax/PLA biocomposite; (b) schematic representa-
tion of the different systems ‘‘layer model’’ studied to probe interfacial
interactions in the biocomposite using model surfaces and colloidal
probes. (Reproduced with permission of Springer Science þ Business
Media from Raj et al.30)
186 Chapter 9
9.2 Materials and Techniques
9.2.1 Preparation of Cellulose Thin Films
One gram of microcrystalline cellulose powder (MW ¼ 70000 g mol–1) was sus-
pended in 50 mL of Milli-Q water for 12 h. This step was followed by two
exchanges for 45 min each with 25 mL of methanol, then four exchanges with
25 mL of dimethylacetamide (DMAc). The activation liquid was removed each
time by using an aspirator pump for each solvent exchange. The filtered swollen
cellulose was dried in a vacuum oven for 24 h at 60 1C. A cellulose solution
(0.5%) was prepared by dissolving the activated microcrystalline cellulose in
9% LiCl/DMAc solvent system.31 A drop of clear cellulose solution was spin-
coated onto a freshly cleaved mica substrate at 3000 rpm for 1 min. The films
were then dried in an oven at 160 1C for 15 min and washed in Milli-Q water to
remove the excess LiCl on the surface and finally dried in a vacuum oven at
100 1C for 15 min. A thickness of B150 nm was obtained for these films.
9.2.2 Preparation of Xyloglucan Thin Films

Xyloglucan was chosen as a model hemicellulose. Traditionally, hemicelluloses
are defined as the alkali soluble materials after the removal of pectic substances
from plant cell walls.32 They are a heterogeneous class of polymers which may
contain pentoses (D-xylopyranose, L-arabinofuranose), hexoses (D-glucopyr-
anose, D-mannopyranose, D-galactopyranose), and uronic acids (D-glucuronic
acid).33,34 The main constituents of hemicelluloses34 are shown in Figure 9.2.
Among the different types of hemicelluloses, xyloglucans (XGs) are the most
predominant hemicellulosic polysaccharides in the primary cell wall, and can
form up to 1.3% of the flax fibre mass.35 Xyloglucans have a backbone chain of
(1-4)-linked b-D-glucopyranose units with a substitution of D-xylopyranose
at position 6. Xyloglucans are classified as XXXG-type or XXGG-type,
Figure 9.2 The main constituents of hemicelluloses. (Adapted with permission of the
American Chemical Society from Hansen and Plackett.34)
Figure 9.3 Chemical structure of the different repeating units of xyloglucan.

Glcp ¼ glucopyranose, Xylp ¼ xylopyranose, and Galp ¼ galactopyranose
units.
depending on the distribution of xylopyranose side-chain units. The XXXGs

have three consecutive backbone residues that are substituted with xylopyr-
anose units and a fourth unbranched backbone residue, while the XXGG
xyloglucans have two consecutive branched backbone residues and two
unbranched backbone residues. Moreover, additional residues are attached to
xylose, depending on the source of xyloglucan. This variation of the structure
dominates the functionality and physicochemical properties. For instance, the
galactose units substituted onto the xylose dominates the water solubility in the
case of xyloglucan extracted from tamarind seed.36 The structures of different
types of xyloglucans36 are shown in Figure 9.3.
A solution was prepared by dissolving xyloglucan (MW ¼ 202 000 g mol–1,
I ¼ 1.8) in Milli-Q water under continuous magnetic stirring during 12 h to
make a 10 g L–1 solution. The solution was then filtered using a 0.1 mm mem-
brane. A drop of solution was spin-coated onto freshly cleaved mica substrate
at 2500 rpm for 1 min. The film was dried in a vacuum oven at 45 1C for more
than 5 h.
9.2.3 Preparation of Pectin Thin Films

Poly(galacturonic acid) (PGUA) was chosen as a model of hydrophilic pectins.
Pectin is a family of complex polysaccharides present in all plant primary cell
walls. The chemical structure of pectin is shown in Figure 9.4.37
The most abundant constituent of pectin in flax fibre is homogalaturonan
(HG), which is a (1-4) linked a-D-galacturonic acid and its methyl ester.35
Some homogalacturonans contain a-D-xylose units. Another constituent pre-
sent in pectin is rhamnogalacturonan I, which contains arabinan, galactan,
and arabinogalactan side chains.38 The PGUA used in this study is a polymer
with a backbone chain containing repeating units of a-D-galacturonic acid
(Figure 9.5).
PGUA in powder form (extracted from orange) was dissolved at a con-
centration of 10 g L–1 in Milli-Q water under continuous magnetic stirring for
12 h. The solution was filtered (0.1 mm) and spin-coated onto a freshly cleaved
mica substrate at 2500 rpm for 1 min. The film was dried in a vacuum oven at
45 1C for more than 5 h.
188 Chapter 9
Figure 9.4 Chemical structure of high-methoxy pectin (DM ¼ 78). (Adapted with
permission of the American Chemical Society from Fischer et al.37)
Figure 9.5 Chemical structure of the repeating unit in poly(galacturonic acid)

(PGUA).
9.2.4 Preparation of PLA Colloidal Probes

Non-crystalline PLA microbeads (PDLLA) were manufactured by an emul-
sion–solvent evaporation process.39 This enantiomer was used to ensure the
lowest roughness at the bead surfaces thanks to its amorphous structure. One
gram of PLA polymer was completely dissolved in 20 mL of CH2Cl2. The PLA/
CH2Cl2 solution (5%) was then poured into a special glass vessel containing 4 g
of poly(vinyl alcohol) dispersant in 200 mL of Milli-Q water. The system was
then continuously stirred for 24 h at room temperature. The PLA microbeads
which had formed were then filtered, washed in deionized water, and vacuum
dried at 45 1C for 3 days.
Colloidal probes were made by gluing a single microbead of PLA at the
extreme end of an AFM cantilever tip. The glue was allowed to cure for 48 h
under ambient conditions.
9.2.5 Atomic Force Microscopy
AFM experiments were performed using a Nanoscope IIIa multimode scanning
probe microscope (Veeco, USA). Experiments under controlled humidity
conditions were performed by placing the AFM inside a homemade glove box
equipped with a hygrometer, with an accuracy of B3%. The temperature inside
the glove box was maintained at 25 1C. A relative humidity (RH) of less than
2% was achieved by keeping dried silica gel in the glove box and passing dry
nitrogen gas through the glove box. The N2 flux was stopped and the system
was equilibrated prior to any AFM force measurements.
Surface imaging of the different polymer thin films was performed in
tapping mode AFM (TM-AFM) under ambient conditions (23 1C and 56%
RH). The silicon tapping mode cantilevers were of the LTESP model with a
nominal tip radius of approximately 10 nm. Direct force measurements
between model surfaces, glued on a magnetic stainless steel disk, and colloidal
probes were performed by cycles of approach and retraction of the piezotube.
The exact value of the cantilever spring constant k was determined to be 51 N
m–1 by the thermal noise method that is based on the thermal fluctuation of
the cantilever. The velocity of the probe was kept constant at 3.5 mm s–1 during
all the experiments and the number of data points was set to 1500. The data
analysis was done using the Nanoscope software V6.13. To improve the
measurement accuracy, statistical analysis of at least 600 force–distance
curves at 10 different arbitrary surface sites were performed. The adhesion
force measured was then normalized according to Derjaguin’s approxima-
tion40 by the radius of curvature of the colloidal probe, determined from
SEM, and plotted as frequency distribution curves of normalized adhesion
force (N m–1).
9.2.6 Microbonding Test

The adherence strength between the flax fibres and the PLA matrix was
estimated by calculating the apparent interfacial shear stress (IFSS) values
obtained from microbond tests on a minimum of 10 pull-out experiments.
A homogeneous PLA polymer microdroplet (length o200 mm) was deposited
on the surface of the flax fibre. In order to achieve this microdroplet, a
microknot was made on the flax fibre with a microfilament of PLA. The set-
up was placed in an oven preheated at 190 1C for 10 min. The sample was
then immediately taken from the oven and quenched at room temperature.
Prior to any tests, the diameter of the fibre near the PLA droplet, the
embedded length of the fibre, and the drop height were measured using an
optical microscope. Pull-out experiments were performed on a tensile testing
machine. On the lower clamp, a homemade X–Y translator with two sharp
knife edges was mounted. The microdroplet was brought just under these
knife edges and the knife blades were brought close together so that the
blades just touched the upper end of the droplet. Tensile loading was applied
at the rate of 0.1 mm min–1.
190 Chapter 9
9.3 Results and Discussion
9.3.1 Model Surfaces
Surface force studies require polymer films that are smooth and homogeneous
over a few micrometres square to ensure reproducible measurements and to
allow for a quantitative determination of the adhesion force using a sphere–
plane model. Figure 9.6 presents AFM images of the different polysaccharides
thin films.
The root mean square (RMS) roughness of poly(galacturonic acid) and
xyloglucan thin films were found to be around 0.5 nm (on a scan size of 25 mm2),
whereas cellulose films present a more important RMS, about 4 nm. The film
thicknesses, as determined by AFM, were all around 100–200 nm. A SEM
image of the PLA colloidal probe (Figure 9.7) shows that the PLA particle
(a) (b)
(c) (d)
Figure 9.6 TM-AFM images (4 mm2 scan size) of thin films of different poly-
saccharides present in flax fibre: (a) xuloglucan, (b) cross-section analysis
of (a), (c) cellulose, and (d) poly(galacturonic acid). (Reproduced with
permission of Springer Science þ Business Media from Raj et al.30)
Figure 9.7 SEM image of a PLA colloidal probe. The scale bar in the picture is 15 mm.
(Reproduced with permission of Springer Science þ Business Media from
Raj et al.30)
is spherical, with an estimated diameter of 30 0,5 mm, non-porous, and

smooth.30
9.3.2 Colloidal Force Microscopy

The principle of colloidal force microscopy has been described elsewhere.17
An example of a force–distance curve measured with CFM is presented in
Figure 9.8.30
In the region of large separation, where the probe and sample do not interact,
the cantilever is not deflected by external forces and the resulting force is equal
to zero (position A). This part is called the ‘‘non-contact region’’. On the
approach stage, as the scanner Z position is increased, the cantilever and the
sample start to interact and a cantilever deflection is induced. The attractive
interaction between the two objects triggers an instability of the probe, which
results in a jump of the probe onto the sample called ‘‘jump into contact’’
(position B). Below this point, for incompressible surfaces a repulsive force
appears and linearly increases with the motion of the piezo-stage. This part
(position C), which corresponds to a straight line in the deflection (or force)
versus piezo displacement (or surface distance) curve, is called the ‘‘constant
compliance region or hard-wall contact’’. After the Z scan has reached a spe-
cified value, the deflection of the cantilever is switched in the opposite direction
(attractive). In the absence of plastic deformation, the deflection observed
during retraction of the cantilever follows the constant compliance region of
the approach (position D). When the position of the cantilever reaches point E,
in which the cantilever spring constant overcomes the adhesion force, the
192 Chapter 9
C) retraction
approach
Cantilever deflection A)
D)
B)
Adhesion F)
Force
E)
Z scanner position
Figure 9.8 Example of force–distance curve showing characteristic features of inter-

actions between two surfaces: (A) non-contact region, (B) jump into
contact, (C) constant compliance region, (D) adhesion event, (E) jump-
out, and (F) return to non-contact region. (Reproduced with permission of
Elsevier from Raj et al.29)
particles and surface separate and the probe ‘‘jumps-out’’ from the surface to
its equilibrium position (position F). This transition is defined as the adhesion
force and can be calculated by simply converting the cantilever deflection to the
corresponding force value according to Hooke’s law (eqn 9.1):
F ¼ kDx ð9:1Þ
where k is the accurate cantilever spring constant and Dx is the cantilever

deflection. For electrically neutral and chemically non-reactive surfaces,
the resulting adhesion force under ambient conditions is the contribution
from van der Waals and capillary forces. Capillary forces due to the for-
mation of a thin layer of water molecules condensed on the surface can
be eliminated by performing the experiments under controlled humidity
conditions.
9.3.3 Direct Force Measurements

At first, the reproducibility of the method and set-ups developed here was
tested by measuring adhesion forces in different parts of different PGUA thin
films, as shown in Figure 9.9a.
It can be observed that good reproducibility is obtained for all the samples
studied, as proved by the small deviation of the centre of the different histo-
grams. For xyloglucan, similar results were obtained, whereas for cellulose a
80
Surface 1: area 1
PLA bead-PGUA
Surface 1 : area 2
56%RH
Surface 1 : area 3
60 Surface 2 : area 1
Surface 2 : area 2
Frequency (%)
40
20
0
0.18 0.20 0.22 0.24 0.26 0.28 0.30
Normalised adhesion force, Fadh/R (N/m)
(a)
40
56% RH
30
Frequency (%)
20
10
0
0.16 0.18 0.20 0.22 0.24 0.26 0.28 0.30 0.32
Normalised adhesion force Fadh/R (N/m)
(b)
Figure 9.9 (a) Frequency distribution of the normalized adhesion force measure-
ments between a PLA colloidal probe and a PGUA film under ambient
conditions measured on different areas of the PGUA film. (b) Frequency
distribution of normalized adhesion forces between a PLA colloid probe
and a PGUA film at 56% RH (N ¼ 600). (Reproduced with permission of
Springer Science þ Business Media from Raj et al.30)
194 Chapter 9
Table 9.1 RMS roughness calculated from AFM images of different
biopolymers.
PGUA, Xyloglucan, Cellulose, PLA,
RMS (nm) RMS (nm) RMS (nm) RMS (nm)
Thin films 0.3 0.3 4 –
(scan area 25 mm2)
Microbeads – – – 1.5
(scan area 4 mm2)
broader adhesion force distribution was obtained. This phenomenon was

attributed to the higher roughness of cellulose films. Once the reproducibility of
the method was verified, a final histogram was plotted from at least 600 single
force measurements on each sample (Figure 9.9b). A quantitative comparison
of adhesion forces is only possible if the films’ roughnesses are comparable,
since adhesion force measurements using the colloidal probe technique are
known to be largely influenced by the roughness of the surfaces or particles in
contact.17 The RMS roughness of all the surfaces used in this study is shown in
Table 9.1.
The roughness parameter, even at the nanometre scale, can affect adhesion
forces between particle and substrate by modifying the contact area and
increasing the distance between the two objects.41 In the present study, the same
bead was used during all the experiments and regularly controlled to avoid any
accidental contamination of the probe.
Figure 9.10a represents the distribution in the normalized adhesion force for
the three different systems obtained under ambient conditions (56% RH).
Under such condition, the adhesion force results from the addition of van der
Waals forces, H-bonds, and capillary forces.17,29 We can observe that adhesion
between PLA beads and the different surfaces is nearly the same, with peaks
centred at around 0.22 N m–1. With respect to the similar RMS characteristics
of the films, the tendency is:
Fadh(PLA/xyloglucan)EFadh(PLA/PGUA) at 56% RH
As previously discussed, the adhesion force obtained for the PLA/cellulose

system must be cautiously interpreted because of the difference in roughness
with the other films. The results obtained here indicate that PLA interacts in a
similar manner with pectins and hemicelluloses, and one might expect that
capillary forces play a major role in adhesion between these materials under
ambient conditions.
In the presence of humidity, it is well known that a condensed water meniscus
between the probe and surface may increase the adhesion force. This additional
force, called the capillary force, is related to the wettability of water to the
substrate as well as to the probe surface. For hydrophobic surfaces, the
thickness of a water film is limited and results in a relatively small capillary
60
RH 56% PLA bead-Cellulose
PLA bead-PGUA
50
PLA bead-Xyloglucan
Number frequency (%)
40
30
20
10
0
0.15 0.20 0.25 0.30 0.35
(a) Normailised adhesion force (N/m)
40
PLA-Cellulose 2%RH
PLA-PGUA PLA-XG
30
Frequency (%)
20
10
0
0.10 0.15 0.20 0.25 0.30 0.35
(b) Normalised adhesion force Fadh/R (N/m)
Figure 9.10 Comparison of normalized adhesion forces between a PLA probe and
different polysaccharide surfaces such as PGUA, xyloglucan, and cellu-
lose under (a) 56% RH and (b) 2% RH. (Reproduced with permission of
Springer Science þ Business Media from Raj et al.30)
force. In opposition, if both surfaces in contact are hydrophilic, the water

capillary bridge between the two surfaces is large and gives an important
additional adhesion force. Experimentally, the contribution of the capillary
force can be eliminated, from the overall adhesion force, by operating the AFM
196 Chapter 9
in vacuum or in a dry environment. Using such experimental conditions,
Burnham et al.42 found a good correlation between the calculated surface
energy, derived from experimental adhesion forces measurements in dry air,
and theoretical values. Similarly, Raj et al.29 obtained a good agreement
between the Hamaker constant for the cellulose/PLA system, as determined
from AFM experiments at 2% RH, and the theoretical value as calculated by
the Lifshitz theory.
Figure 9.10b shows the effect of RH on pull-off forces for the different sys-
tems. When the RH is decreased to 2%, the ‘‘PLA/cellulose’’ adhesion peak
shifts to a lower value, whereas in the case of PLA/PGUA and PLA/xyloglu-
can, no variation is observed.30 As already reported in the literature, when the
RH tends to zero, the adhesion force remains simply ensured by van der Waals
forces and H-bonds without capillary forces and therefore decreases.43,44
Surprisingly, for PGUA and xyloglucan films, no such effect is observed. This
failure to observe the humidity dependence has already been reported,45 but it
was specific for hydrophobic surfaces where the amount of water is limited. The
roughness of the surface is also an important parameter to consider, since the
contact angle of water on a rough surface is different from that on a smooth
surface and consequently affects the magnitude of the capillary force.46 In our
case, the main contribution of the capillary force does not come from the
difference in roughness between the films but rather in their characteristic
surface energies, as revealed by contact angle measurements. Water contact
angles on cellulose, xyloglucan, and PGUA films are respectively equal to
40 21, 25 11, and 13 11. Xyloglucan and PGUA are more hydrophilic
surfaces and smoother than cellulose films and, as a consequence, are hydrated
and retain water molecules even at low RH. Therefore, we believe capillary
forces are important in these systems and several assumptions can be proposed
to explain the non-decrease of the adhesion force with RH. The first one is
based on the remaining water inside PGUA and xyloglucan films, which are
known to influence the polymer film structure and properties. We are reminded
here of the important role of water molecules in the fibre structure to ensure
optimized cohesion between its various entities. Hydrogen bonds are known to
be important in vegetal fibres.47 Moreover, it is well known that water, acting
as a plasticizer, affects the glass transition temperature (Tg) of amorphous
hydrophilic polymers.48,49 Water molecules yield a large increase in mobility,
due to increased free volume and decreased local viscosity. Water molecules
plasticize polysaccharide films and therefore decrease the polymer modulus
through the reduction of its glass transition temperature, for example.50
For instance, an amount of water of 0.1 g L–1 will decrease the Tg of pectin
from 45 to 0 1C.
In the JKR theory, the contact area (ac) is given by (eqn 9.2):
1=3
3pR2 Wadh
ac ¼ ð9:2Þ
2K*
where R is the radius of the probe, Wadh is the work of adhesion, and K* is the
reduced elastic modulus. It can thus be observed that if K decreases, the contact
area between the two objects will increase and so consequently will the mea-
sured adhesion force. Since water molecules are difficult to remove from polar
PGUA and xyloglucan, the measured CFM adhesion force will not decrease
from 52% to 2% RH. Some efforts have to be done to measure the water
content inside polysaccharides films.
A second assumption is based on the fact that other adhesion mechanisms,
independent of the RH, may also be present in the interaction mechanisms
between PGUA, xyloglucan, and PLA. The ‘‘diffusion theory’’ of adhesion is
accounted for by the time and temperature dependence of the adherence
between miscible, soft polymers.51,52 In practice, the inter-diffusion process
requires sufficient chain mobility, which can, of course, be related to the glass
transition temperature of the polymers.53 There is, to the best of our knowl-
edge, no evidence of such a mechanism in biocomposites and further experi-
ments should be conducted to verify this hypothesis.
9.3.4 Interfacial Shear Stress and Work of Adhesion

Interfacial shear stress (IFSS) at the flax/PLA interface was estimated from the
microbonding pull-out test using the Kelly–Tyson equation (eqn 9.3):
Fmax
ti;mean ¼ ð9:3Þ
pdle
where Fmax is the maximum pull-off force at debonding, d is the fibre diameter
and le is the fibre embedded length inside the polymer droplet. The above
equation assumes that the force Fmax at the instant of debonding is predicted to
be directly proportional to the joined surface area between the fibre and the
matrix, and the droplet shears off from the fibre surface when the average shear
stress at the interface, ti,mean, becomes large enough to break the interface. The
apparent shear stress was determined from the linear regression of the plot of
debonding force versus bonding area.
The work of adhesion at the flax/PLA interface was estimated, on the one
hand, from contact angle measurements according to the Young–Dupré
equation (eqn 9.4):
Wadh ¼ gl ð1 þ cos yÞ ð9:4Þ
where gl is the interfacial energy of the PLA melt drop that makes a contact
angle of y with flax fibre’s surface. Also, the work of adhesion was determined
from colloidal force measurements between a PLA bead and different
198 Chapter 9
polysaccharide surfaces. The work of adhesion for two surfaces in sphere–plane
geometry contact can be estimated according to the Johnson–Kendall–Roberts
(JKR) theory (eqn 9.5):54
Fadh 3
¼ pWadh ð9:5Þ
R 2
and the Derjaguin–Muller–Toporov (DMT) theory (eqn 9.6):55
Fadh
¼ 2pWadh ð9:6Þ
R
The JKR theory considers that adhesion takes place inside the area of con-
tact and can be applied in cases of large tips and soft samples with a large
adhesion. JKR theory is thus more suited to model colloid probe microscopy
experiments where we use a large tip (colloidal probe) and a soft polymer film.
On the other hand, the DMT theory takes into account the adhesion outside
the contact area and is applied in cases of small tips and stiff samples with small
adhesion.17
The Maugis parameter (l) provides a useful measure of which model is
appropriate and is given by eqn (9.7):56
sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
3
Rg2
l ¼ 2:06 2 D3
ð9:7Þ
pEtot 0
where Etot is the reduced elastic modulus of the two surfaces and D0 is
the separation distance between the two materials. In cases where l41, the
JKR model is valid, whereas if lo1, the DMT model is used. In our case, we
found the Maugis parameter, l, close to 100, thus justifying the use of the JKR
model.
The work of adhesion, as measured by AFM and contact angle measure-
ments, are shown in Table 9.2.
Table 9.2 Comparison of work of adhesion (Wadh) estimated for different

systems from contact angle, and CFM experiments.
CFM, JKR Wadh CFM, JKR Wadh Wadh contact
System (56% RH) (mJ m–2) (2% RH) (mJ m–2) angle (mJ m–2)
Flax fibre/PLA – – 60.78 ( 2.8)
(y ¼ 55.6 41)
PLA bead/cellulose film 40.3 ( 3.0) 30.6 ( 1.9) –
PLA bead/xyloglucan film 42.5 ( 1.7) 42.5 ( 1.7) –
PLA bead/PGUA film 46.7 ( 1.9) 46.7 ( 1.9) –
9.3.5 Verifying the Nardin–Schultz Relation for Flax/PLA
Biocomposites
The semi-empirical relationship between IFSS (t) and the work of adhesion
(Wadh) proposed by Nardin and Schultz15 is shown in eqn (9.8):
1=2
Em Wadh
t¼ ð9:8Þ
Ef d
where Em and Ef are the elastic modulus of the polymer matrix and the fibres,
respectively, and d is the distance equal to 0.5 nm that corresponds to an
equilibrium intermolecular centre-to-centre distance involved in molecular
interactions.
We tried to verify the Nardin–Schultz model for flax fibre/PLA biocompo-
sites, using our experimental data. To plot these experimental points, the elastic
modulus of the fibre and PLA matrix were taken as Ef ¼ 65 GPa57 and Em ¼ 3.6
GPa,58 respectively. These data, which represent macroscopic mean values for
these materials, are well established in the literature. Figure 9.11 plots the
Nardin–Schultz relationship between IFSS and Wadh for flax/PLA bio-
composites, and compares that with the linear relation experimentally verified
for model composite materials.
It can be observed that for flax/PLA composites the work of adhesion cal-
culated from the contact angle technique did not fit the Nardin–Schultz linear
250
Nardin-Schultz relation for model composites
Wadh: Contact angle (Flax fibre-PLA)
Wadh CFM: (PLA bead-PGUA film)
200 Wadh CFM: (PLA bead-XG film)
Wadh CFM: (PLA bead-Cellulose film)
(Ef /Em )(1/2) τapp (Mpa)
150
100
50
0
0 20 40 60 80 100 120
Wadh (mJ/m2)
Figure 9.11 The Nardin–Schultz relationship between IFSS and Wadh obtained for
flax/PLA biocomposites.
200 Chapter 9
relation. However, when the work of adhesion was calculated from more local
techniques such as CFM, we observed that the points were placed close to the
Nardin–Shultz linear relation. It can be seen that the closest agreement was
achieved for the work of adhesion calculated for the PLA/cellulose system. In
an earlier work, we have shown the contribution of van der Waals forces for
PLA/cellulose interactions by calculating the Hamaker constant from pull-off
forces.29 In the Nardin–Shultz model the work of adhesion between two solids
is considered to be the contribution from dispersive interactions (London
forces) and electron acceptor–donor interactions (Lewis acid–base interac-
tions).15 However, in cases where other adhesion mechanisms, e.g. covalent
bonding, mechanical interlocking, or interdiffusion processes, may take part in
interfacial adhesion, then the Nardin–Schultz relation cannot be directly
applied. This was clearly evident in the case of the hydrated polysaccharides
poly(galacturonic acid) and xyloglucan, where the work of adhesion with PLA
was far from simple molecular interactions and involved complex mechanisms
such as capillary forces and interdiffusion phenomena (see Section 9.3.3). The
deviation from the Nardin–Schultz linear relation in the case of hydrated
polysaccharides (Figure 9.11) could be attributed to the complimentary
‘‘adhesion mechanisms’’ that exist at the interface with these materials.
9.4 Conclusion
Colloid force measurements using AFM have been successfully tested to
directly probe interfacial interactions in flax fibre/PLA biocomposite materials.
These direct force measurements, using smooth polysaccharides films and PLA
colloidal probes, allows for qualitative comparison between different systems
and provides an insight into the nature of complex interactions involved at the
biocomposite interface. Adhesion forces were shown to be dominated by
capillary forces under ambient conditions. Force measurements under con-
trolled humidity conditions show that PLA/cellulose is probably the weak
interface in the biocomposite in terms of interactions. It also underlines the
important role of water in the adhesion between the PLA and some hydrated
polysaccharides such as pectin and hemicellulose. Even at low RH, the adhe-
sion force was not reduced for these polymer films. We suggest that the large
adhesion, and its insensitivity of as a function of RH, was due to the high
amount of water within the film, which presents low elastic modulus, and hence
may deform locally upon contact with the PLA bead to give an increased
contact area. Interdiffusion mechanisms may also participate in the overall
adhesion mechanism between these polymers. The importance of water was
emphasized and is clearly interesting since it should be considered for real
systems.
The Nardin–Schultz linear relation between work of adhesion and interfacial
shear stress was tested for flax/PLA biocomposites. The work of adhesion
estimated from colloidal force measurements using AFM was in better agree-
ment with the Nardin–Schultz linear relation, when compared to the Wadh
calculated from global and averaging techniques such as contact angle mea-
surements. A best linear fit was observed for the PLA/cellulose system, where
the work of adhesion was dominated by molecular interactions such as van der
Waals forces. However, for systems in which interfacial adhesion is dominated
though complex interaction mechanisms, a slight deviation from the Nardin–
Schultz linear relation was observed. This was typically the case observed for
systems involving hydrated polysaccharides such as pectin and hemicellulose,
where interfacial interactions were dominated through a combination of
complex ‘‘adhesion mechanisms’’ involving capillary forces and interdiffusion
phenomena.
Acknowledgement
Financial support for this work from Region Bretagne, Cap Lorient (France),
is deeply acknowledged.
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CHAPTER 10
Zein: Structure, Production,

Film Properties and Applications
NARPINDER SINGH,*a SANDEEP SINGH,a
AMRITPAL KAURa AND MANDEEP SINGH BAKSHIb
a
Department of Food Science of Technology, Guru Nanak Dev University,
Amritsar, 143005, India; b Department of Chemistry, Wilfrid Laurier
University, Science Building, 75 University Ave. W., Waterloo,
Ontario N2L 3C5, Canada
10.1 Introduction
Alcohol-soluble storage proteins of corn are also known as prolamines and are
generally referred to as zein. Zein is a naturally occurring protein polymer
obtained from corn gluten meal (CGM), a co-product obtained during the wet
milling of corn to produce cornstarch. Zein is also produced as a co-product of
the ethanol industry. Whole corn grain contains around 39% zein and the
endosperm portion has about 47%. CGM contains around 60–74% protein,1
20% starch and 4% oil. Zein constitutes one-half or more of CGM proteins;
therefore CGM is the main raw material for both commercial and laboratory
scale production of zein. Zein proteins account for about 60% of the total
protein of corn grain. Zein contains 21.4% glutamine, 19.3% leucine, 9.0%
proline, 8.3% alanine, 6.8% phenylalanine, 6.2% isoleucine, 5.7% serine and
5.1% tyrosine.2 Zein is deficient in essential amino acids, such as lysine and
tryptophan, which makes it poor in nutritional quality. Zein has a high pro-
portion of non-polar amino acids (mainly leucine, alanine and proline),

204
Zein: Structure, Production, Film Properties and Applications 205
3
residues that are responsible for its poor solubility in water. Both its insolu-
bility in water and poor nutritional properties reduce its applications in human
food formulations.
The present chapter deals with the isolation, structure, film forming prop-
erties and applications of zein.
10.2 Structural Characteristics

Zein is classified into four classes, viz. a, b, g and d, according to their solubility.
Among the four fractions, a-zein with a molecular weight of 21–25 kD is the
most abundant fraction. The b-, g- and d-zein fractions have molecular weights
of 17–18 kD, 27 kD and 9–10 kD, respectively.4,5 These fractions of zein differ
in their amino acid sequences and surface charges. Paulis et al.6 reported that
35% of the total zein was a-zein, which had two major bands with molecular
weights of 24 kD and 22 kD and an amino acid and peptide composition similar
to whole zein.6–9 Zein varies in composition and concentration depending upon
corn genotypes,5,10 location of protein bodies in the grain11,12 and maturity.13,14
a-Zein has large amounts of hydrophobic residues such as leucine, proline,
alanine and phenylalanine,15 which contribute to its hydrophobic properties.
The average hydrophobicity of zein was reported to be 50 times larger than
albumin, fibrinogen, etc., and hence a higher percentage of an alcohol/aqueous
mixture is required for the dispersion of zein.16 When dissolved in aqueous
alcohol, the protein was reported to be rich in a-helical content (50–60%), with
turn or random coil configurations comprising the remaining structure.17
Several models have been proposed for the 3D structure of a-zein, including
one composed of a series of anti-parallel helices clustered within a distorted
cylinder.17 Argos et al.17 proposed a structural model for a-zein using optical
rotatory dispersion and circular dichroism measurements (Figure 10.1). The
model consists of repeat units forming an a-helix and having three polar seg-
ments on its surface (Figure 10.1a). The a-helices interact with the others by
hydrogen bonds using two of the three polar segments (Figure 10.1b), while the
third polar segment is utilized for intermolecular contacts (Figure 10.1c). a-Zein
behaves in solution as asymmetric particles with an axial ratio from 7:1 to 28:1
as indicated by ultracentrifugation, birefringence, dielectric and viscosity
experiments.18 a-Zein has coiled-coil tendencies, resulting in a-helices with about
four residues per turn in the central helical sections, with the non-polar residue
side chains forming a hydrophobic face inside a triple superhelix.18 The nine
helical segments of the 19 kD protein were modeled into three sets of three
interacting coiled-coil helices, with segments positioned end-to-end. The
resulting structure lengthens with the addition of the N- and C-terminal sections,
to give an axial ratio of B6:1 or 7:1. Structural studies using different techniques
such as circular dichroism, IR, NMR and optical rotatory dispersion suggested
that Z19 (19 kDa) has B35–60% helical character, made up of nine helical
segments of about 20 amino acids with glutamine-rich ‘‘turns’’ or ‘‘loops’’.17,18
Garrett et al.19 developed a model based on hydrophobic membrane pro-
pensities and helical ‘‘wheels.’’ It was suggested that pairs of repeats formed
206 Chapter 10
(a) (b)
(c)
Figure 10.1 Proposed model of zein structure: (a) helical wheel for the 18-residue
consensus repeat sequence showing three polar regions; (b) nine-helical
zein protein structural model; (c) arrangement of zein proteins within
a plane as well as for the stacking of molecular planes. Q ¼ glutamine
residues.17
anti-parallel a-helical hairpins and were arranged in a hexagonal net. X-ray

scattering and viscosity studies have demonstrated that a-zein in solution is a
relatively rigid asymmetric particle with a diameter between 0.7 and 1.5 nm and
a length of B15 nm by small-angle X-ray scattering (SAXS) and 24 nm by
viscosity.20 Matsushima et al.21 suggested another three-dimensional model
consisting of nine (Z19) or ten (Z22) helical segments folded in anti-parallel
fashion linked by glutamine-rich turns and held in place by hydrogen bonds.
They proposed that the helical segments were aligned to form a compact
elongated prism-like shape rather than an open ring. Such an arrangement
allows a-zein molecules to contain different repeat units with greater flexibility
and enhanced intramolecular hydrogen bonding. Momany et al.18 favored the
stick model (Figure 10.1) to represent the N-terminal segment of Z19. The
resulting compact structure indicated that the cysteine residue (C27 or C6 in
the renumbered sequence starting at residue 22) is exposed on the surface
and therefore is available to form a disulfide bridge with the cysteine residue of
the Z22 (22 kDa) protein, creating the naturally found dimer. The N- and
C-termini are close to one another, a result of a large chain reversal in the
middle of the segment and the disruption of secondary structure because of the
nine proline residues in the segment.
10.3 Isolation and Production

Various methods are used for the manufacture of zein. These methods differ in
terms of the raw material, the solvent used for extraction, purification and the
method used for zein recovery. Almost the entire zein present in raw corn comes
in the CGM, which is obtained as a co-product during the wet milling of corn.
About one-half or more of the protein in CGM is zein. CGM is a low-cost raw
material for the production of zein and is primarily used as an animal feed. The
quality of CGM varies widely in protein content, depending upon the method of
corn processing. The recovery of zein from CGM also depends upon its handling
and processing conditions. For example, recovery of zein from CGM decreases
with an increase in drying temperature.22 Raw dry-milled corn gives a low zein
yield when used as starting raw material23 and leads to a higher cost of recovery.
A low recovery of zein makes the process uneconomical, unless the methods used
for solvent recovery and concentration of zein are efficient and cost effective.24
Distiller’s dried grains with ‘‘solubles’’ (DDGS) is another source of zein proteins
and is obtained as a co-product during a dry-grind ethanol process. The dry-grind
ethanol process involves corn grinding followed by saccharification and fer-
mentation of glucose to produce ethanol, leaving concentrated fractions like
cellulosic materials and proteins. DDGS has a protein content of around
27–30%. The quality and composition of the proteins present in DDGS varies
widely, depending upon the drying and processing conditions. Drying at higher
temperature can induce cross-linking of zein proteins and bring about changes in
functional properties. DDGS is not a preferred raw material for producing high-
quality zein since higher temperatures are used during drying.25,26
Zein contains predominantly nonpolar amino acids and therefore the sol-
vents used for the extraction of zein need to possess wide characteristics,
containing both ionic and non-ionic polar groups as well as nonpolar groups,
either structure-wise (in the case of pure solvents) or composition-wise
(for mixed solvents). Zein is considered ‘‘soluble’’ if more than 0.5% (w/v) of
the protein dissolves in the solvent and produces a clear solution at room
temperature. Most of the zein extraction processes involve the use of two
solvents in succession: a polar solvent (aqueous solutions of ethanol or
208 Chapter 10
isopropanol) and a nonpolar solvent (hexane or benzene) for the removal of
lipids and pigments. The solubility of zein in ethanol varies between 2 and
60% (w/w) at constant temperature, and varies with the ethanol concentration.
At lower (o40%) and higher (490%) concentrations of ethanol, two liquid
phases appear; both contain zein, water and ethanol. This phenomenon is
widely used to recover zein after extraction from CGM. Zein has two major
fractions, a- and b-zein, as described by McKinney.27 The a-zein is soluble in
95% ethanol while b-zein is soluble in 60% ethanol and insoluble in 95%
ethanol. The a-zein has a lower content of histidine, arginine, proline and
methionine than the b-zein. The b-zein is relatively unstable, and precipitates
and coagulates readily, and hence is not a constituent of commercial zein.
A variety of laboratory-scale methods has been studied and described in the
literature wherein zein has been fractionated by precipitation, cation exchange
chromatography,28,29 differential solubility,5,30,31 cryo-precipitation,32 charcoal
and gel filtration.33–35 Zein isolation by precipitation involves adding water to
solutions of zein in ethanol or by using cellosolve.31,36 Extraction of zein by the
differential solubility method37 from CGM gives yields up to B90% with iso-
propanol (80%), followed by a petroleum ether treatment. Cation exchange
chromatography28 involves use of defatted ground corn and extraction with 70%
ethanol. The corn extract is refined using an Amberlite IRC-50 column with a
NaCl gradient, which gives a yield of B86%. Gel filtration chromatography33
was used to separate zein from corn extract and processed by a combination of
LH-20 purification and carbon adsorption, resulting in 90% purity.
Commercial methods for the production of zein from corn have been
reviewed by Shukla and Cheryan38 and are covered in several US patents. Most
of the economically viable methods use aqueous solutions of ethanol or iso-
propanol (IPA) for the first extraction of zein. IPA is the preferred solvent for
zein extraction as it has more affinity than ethanol in terms of zein con-
centration, i.e. a higher zein concentration at the same solvent-to-solids ratio.
The subsequent separation (e.g. hexane extraction for pigment and oil removal)
is better because the separation between zein and the hexane layer is easier. IPA
dissolves in the hexane layer to a lesser extent, which results in lower distillation
costs. These common solvents extract several other components (xanthophylls,
polyamines, lipids, etc.) along with zein. Hence, various commercial processes
or patents are distinguished based on methods of separation and purification to
increase the purity of the zein.
Zein was manufactured on a large scale by Corn Products Corporation (CPC)
from 1939 to 1967 and the method of purification by precipitation of zein has
been described in patents.39,40 In the CPC process (Figure 10.2.),41 dried or wet
CGM was contacted with hot 86–88% IPA or 93–95% ethanol at high pH
and temperature (50–60 1C) in either a batch or continuous extractor for about
1.5–2 h. The use of reducing agents has been mentioned in the literature,42 but are
not commonly used in any industrial process. The extract was filtered and/or
centrifuged and the filtrate, containing zein (B6% w/v) and impurities, was then
clarified by standing or vacuum filtration43 and followed by cooling (15 1C).
Swallen44 patented a process of extraction of zein from CGM using 85% IPA in a
HEXANE CHILLED
1:1 WATER
Corn Gluten Meal

+ CENTRIFUGE
95% EtOH / 86% IPA ZEIN
PRECIPITATION
SOLVENT TANK
MIXER
Spent 24% FLASHER
6% zein &
Gluten zein
SEPARATOR
STEAMED 15°C FILTRATION
Hexane, Oil & Pigments GRINDING

VACUUM
FILTER
Residual Meal SOLVENT DRYING
RECOVERY
ZEIN
Figure 10.2 CPC process for production of zein from corn gluten meal.
solute-to-solvent ratio of 1:3.5 at 60 1C. Fats and colour pigments were extracted
from zein by using solvents like hexane or benzene. The zein was finally
precipitated either using excess amounts of cold water or at low temperatures
(–15 to –25 1C). It was then vacuum dried and ground to produce zein of a light
yellow colour. The CPC process was modified to include alkali treatment, where
the pH was raised to 12 with NaOH and held for 30 min for deamidation of amino
acid residues.2 The pH was subsequently lowered with HCl and zein precipitated
in cold water. This procedure improved the stability and gelation properties of
zein.45 The CPC process had some major disadvantages, such as high operating
costs due to its complex solvent recovery systems (primarily distillation), low yield
and high solvent losses during extraction.46,47 Gelation of zein due to the varia-
tion in the pH of the solution and the quality of final product were also some of
the other shortcomings of this process.
A process by Carter and Reck48 known as the ‘‘Nutrilite process’’ involved
extraction of zein from CGM using 88% aqueous IPA in a solute-to-solvent
ratio of 1:4, with 0.25% NaOH for 1 h at 55–65 1C (Figure 10.3). Precipitation
of the zein solution was carried out by chilling to –15 1C. Zein produced by this
method contains 2% oil and xanthophylls, which can be partially removed by
re-extraction with 88% IPA and subsequent chilling. Zein prepared in this way
contains less oil (0.6%) and a lighter colour. This process appears to control
gelation problems because low temperatures impart stability against dena-
turation of zein. However, the process was expensive since large volumes of
solvent and low temperature precipitation were required.
Cook et al.49 patented a process for the purification of zein and removal of
pigments for pharmaceutical use. The CGM was made free of starch and
washed several times with absolute ethanol to remove pigments and oil.
The CGM was then washed with water and extracted with 80% ethanol. The
extract was treated with activated carbon to remove further flavors and
210 Chapter 10
Supernatant 88% Supernatant

(4 volumes) IPA (6 volumes)
CORN GLUTEN
MEAL SETTLING
–15°C TANK SETTLING
+ MIXING
88% IPA (4:1) TANK
TANK
Spent
Gluten
CENTRIFUGE Zein-Alcohol Layer –15°C
(30% Zein) Zein-Alcohol Layer
STEAMED DRYING
DRYING
12% zein
GRINDING
VACUUM GRINDING
Residual Meal 15°C
FILTER
Zein
Zein (0.6% Oil)
(2% Oil)
Figure 10.3 Nutrilite process for the production of zein from corn gluten meal.
pigments, precipitated with water and finally dried. The method extracts zein
devoid of flavor and pigments; however, a large amount of ethanol is used with
about B2% of zein lost during the extraction of pigments. The removal of odor
improves zein’s marketability for applications such as gum.50
A method for refining zein after extraction was patented by Takahashi and
Yanai.51 Zein was extracted from CGM using aqueous acetone (70% v/v) in a
respective ratio of 1:5 at 40 1C for 4 h. The solution was extracted and con-
centrated by evaporation to a syrup-like consistency, followed by precipitation
using absolute acetone. The method gave higher zein yield with minimal colour
than the zein produced by the method of Carter and Reck.48
In most of these methods, organic solvents, high pH, enzymes, etc., were used
for zein extraction and had some effect on the native conformation and com-
position of the proteins, which limit its food and non-food applications.
10.4 Film Forming

Zein is chemically inert and its globular structure poses difficulties in molding
into different articles. Zein film formation occurs through development of
hydrophobic, hydrogen and limited disulfide bonds between zein chains.52
Films made from zein alone are brittle and plasticizers are incorporated to
make them soft and permanently flexible.45,53–57 Parris and Coffin58 prepared
unplasticized casting zein films in aqueous ethanol or acetone. The tensile
properties indicated that films prepared in acetone were stronger but less
flexible than those prepared in ethanol. Both types of films, however, were too
brittle for most applications. Plasticizers are low molecular mass organic
compounds added to soften rigid polymers.59 These compounds act by
reducing the glass transition temperature of polymers, reducing their crystal-
linity or melting temperature.60 Plasticizers dissolve in the polymer that sepa-
rates the chains from each other and thus facilitate molecular movement. These
are applied to increase workability, flexibility and extensibility of polymers.61
Plasticizers also act as internal lubricants by reducing frictional forces between
polymer chains,62 and their action may bring changes in the mechanical
properties of polymers.63,64 Plasticizers cover a wide range of chemical com-
pounds, including esters, hydrocarbons, water, alcohols, glycols, phenols,
ketones and ethers.62,63 A plasticizer is considered ideal if miscible and com-
patible in wider proportions with plastic components. Generally, compatibility
and permanence are said to follow opposite trends; therefore, the more com-
patible and efficient is a plasticizer, the more rapidly it can diffuse out of the
polymer.63 Lai and Padua65 indicated that the addition of palmitic acid at a
ratio of 0.5 g g1 of zein or stearic acid to 0.25 g g1 of zein substantially
increased the tensile strength of the zein sheets. Glycerol has also been tried as a
plasticizer in zein films.66,67 However, Park et al.68 reported that the interaction
between protein molecules and glycerol is weak, and excess glycerol easily
migrates through the film matrix. The processing techniques such as kneading,
blowing and/or extrusion, used for film formation, could strengthen the plas-
ticization effect of free fatty acids in zein films.65,69 Therefore, some input
energy to promote the interactions between fatty acid plasticizers and zein
molecules is required during processing.70
Zein films synthesized through an acylation reaction with lauryl chloride
were reported to be more flexible and tougher than pure zein. The hydro-
phobicity on the surface was further increased because of the attached alkyl
chains, which could potentially improve the water barrier properties of these
zein-based biomaterials. This study suggested a great potential of acylated zein
as food packaging and coating materials, with improved properties. Ghan-
barzadeh et al.71 reported that the films plasticized by galactose had relatively
higher tensile strength than films containing fructose and glucose. This could be
attributed to a more hygroscopic nature of zein films containing glucose and
fructose than those containing galactose. Zein films are generally formed by a
wet-and-dry process. Film formation involves dissolving zein, plasticizers and
other agents in an appropriate solvent and the solution is cast on a flat nonstick
surface. The films are peeled off after the solvent has evaporated. Another
method involves the preparation of a moldable resin of zein and plasticizers,
which are later formed into a film by a heat process.65,72 Resin zein films are
obtained by stretching the resin over circular rims or pressing in a hot press.
10.5 Film Properties

Formation of zein films for food packaging has been extensively reported. The
tensile strength of zein films was reported to be comparable with wheat gluten
films.73 Singh et al.66 reported tensile strength and strain at failure of 27 MPa
and 0.029, respectively, for zein–glycerol films. Alcohol-soluble proteins, such
212 Chapter 10
as zein, wheat gluten and fish myofibrillar proteins, form films of relatively low
water vapor permeability (WVP) when compared to other proteins.74 Such
proteins are required to be modified by different methods to improve their
functional properties. Films made from zein acetate have higher water resis-
tance, strength and flexibility.75 The major problem encountered during mod-
ification of zein is its lower solubility. Zein readily dissolves in an ethanol/water
(90:10 w/w) mixture. However, this solution poses difficulty during chemical
modification, since alcohol/water readily reacts with electrophilic reagents
before zein does. To overcome this, the use of N,N-dimethylformamide as an
inert solvent was suggested for zein modification.76 Plasticization affects the
WVP: zein films without plasticizers had lower WVP values than those plas-
ticized with glycerol or mixtures of glycerol with poly(ethylene glycol) or
poly(propylene glycol).53,68,77 Protein films have low oxygen permeability
at low to intermediate relative humidity. Oxygen permeability values were
comparable to those of modest oxygen barriers, such as polyesters, and
approaching those of the best oxygen barriers, i.e. ethylene–vinyl alcohol
copolymer and poly(vinylidene chloride).74 Low molecular weight molecules
such as some polyols and fatty acids could be used as plasticizers to improve the
mechanical properties of zein films.78 However, phase separation as a result of
immiscibility between zein and such plasticizers take place; this usually results
in instability in the mechanical properties and often heterogeneous surfaces.
Another disadvantage of using polyol plasticizers is that they exacerbate the
sensitivity of zein-based materials to moisture.79–81 Thus, new techniques to
improve the water resistance and mechanical properties of zein-based materials
need to be developed. Oleic acid has also been used to plasticize zein-based
films.45,65,82 Zein films plasticized with oleic acid were reported to exhibit tensile
and moisture barrier properties that make them potentially useful as biode-
gradable packaging materials.65,83 Zein films prepared with drying flax or tung
oils showed higher resistance to water vapor.84 Santosa and Padua69 also
evaluated the plasticizing effect of oleic acid on zein film characteristics. The
WVP of zein films was observed to be lower than that of ethyl- and methyl-
cellulose films. Zein films have lower storage stability and become discoloured
and oxidized. Zein sheets prepared using oleic and linoleic acids as plasticizers
resulted in an increase in the elongation percent, while Young’s modulus and
water absorption decreased.69 Linoleic acid was more effective than oleic acid
in reducing the water absorption of sheets, possibly due to the filling up of the
pores and gaps in the structure and preventing it from swelling.69
10.6 Modifications
The zein structure is modified by physical and chemical methods to enhance its
functional properties such as water solubility, foaming and emulsification.85,86
Moderate modification of zein structures by enzymatic and chemical treatments
is desired for partially unfolding zein molecules to improve the functional
properties. The modifications of zein are done to the desired level because
excessive and uncontrolled modifications lead to reduce functionality87,88 due
89
to fragmentation and truncation of the zein backbone. Protein structures are
influenced by many factors such as temperature, concentration, salt type, pH
and solvent type. High temperature (90 1C) results in denaturation of secondary
and tertiary structures, along with oligomerization of the zein.85 Cross-linking
induced by chemical reagents such as formaldehyde, glutaraldehyde, epi-
chlorohydrin, citric acid, butane-1,2,3,4-tetracarboxylic acid, polymeric dia-
ldehyde starch, 1,2-epoxy-3-chloropropane and dialcohols between zein
molecules has also been reported. The cross-linking of zein molecules caused a
significant increase in tensile strength of zein films.58,89–91 It was also reported
that cross-linking of zein molecules with 1-(3-dimethylaminopropyl)-3-ethyl-
carbodiimide hydrochloride and N-hydroxysuccinimide improved film forming
properties while the aggregation phenomenon in solution was suppressed.92 Wu
et al.93 used polycaprolactone (PCL) to modify zein, where the hydrophobic
chain of PCL besides cross-linking the zein molecule also serves as a plasticizer
and significantly increases the elasticity of zein sheets.
Different cations (i.e. Na1, K1, NH41 and Mg21) have also been reported to
strengthen zein structure selectivity and interaction with non-ionic surfac-
tants.94 Methanol, ethanol and isopropanol have also been tried to modify the
morphology, diameter and distribution of fibres from the electrospinning of
zein.95 Cabra et al.85 reported that different pH values (0.5–2 N HCl or NaOH),
with or without applying heat, would induce zein deamidation, resulting in an
improved emulsifying property. Under similar pH conditions (0.05 N HCl with
heat), Chiue et al.87 found a decrease in zein’s fat-binding capacity, leading to
the loss of radical scavenging ability. Bakshi et al.96 blended zein films with self-
assembled (SA) bovine serum albumen (BSA) conjugated nanoparticles (NPs)
to improve the mechanical properties of zein films. BSA conjugated NPs were
produced through biomineralization of a gold salt by BSA. SA NPs promote
the mechanical properties of zein films, as hydrophobic proteins of zein have
preferential interactions with predominantly hydrophobic SA NPs. Colloidal
SA NPs in the form of soft films thus provide better orientations for unfolded
zein to produce strong and flexible protein films.
10.7 Applications
Zein is generally recognized as safe (GRAS) and is biodegradable97,98 and
biocompatible.99,100 Some applications include free-standing packaging mate-
rial,101 emulsifier,85,86 chewing gum base,102 antioxidant,87,103 etc. Many of
these applications (chewing gum base and antioxidant) are due to higher ali-
phatic indexes, surface hydrophobicity17,85 and the high fatty acid-binding
capacity88 of zein. In the food and confectionary industries, zein is being used
for coating enriched rice, candies such as chocolates and jellybeans, dried fruits,
nuts, nutmeats, and for the encapsulation of flavors and sweeteners. It is used
as a coating on bakery products as a vegetable protein. Other packaging
applications include the use for frozen foods and ready-to-eat chicken.104
Some other applications include tablet coating, drugs encapsulation105,106
and as a tissue scaffold.107,108 Singh et al.66 reported that zein iodine solution
214 Chapter 10
could be used as a film forming medication (ointment) when high anti-micro-
bial activity against pathogenic microorganisms or infections is required. Such
ointments have the potential to replace substantially oily ointments that are
inefficient and might spoil or stain clothes. Zein films containing salicylic acid
and acetylsalicylic acid with or without glycerol for structural, mechanical and
dissolution properties were also evaluated.67 This study reported that random
coils, a-helix and b-sheets mainly governed the secondary structure of zein,
depending on glycerol and the level of model molecules. It was suggested that,
from a bio-medical perspective, zein films could act as drug release devices
provided a thorough understanding of the interactions between zein proteins
and the active ingredients be established.
Zein has a wide variety of other non-food applications, such as the manufacture
of plastics, fibres, adhesives, coatings, binders, etc. Films produced using zein have
excellent water barrier properties because of its hydrophobic characteristics and
stability in acid and alkaline solutions.52 Therefore zein has been employed as a
raw material for waterproofing paper, coating wood, adhering plywood, damp-
proofing and tablets.45 The fibres made from zein are used for garments, hats and
other commercial applications.109. Zein fibres produced by electrospinning have
been suggested to have better applications in the fibre market.95,110
Chen et al.111 produced zein nanofibres by the electrospinning of aqueous
alcohol solutions. The electrospinning process produced nanoparticles, nanofibre
mats or ribbon-like nanofibre mats and they concluded that fibre morphology
was primarily affected by solution concentration. Zein electrospun nanofibre
mats were flexible and lustrous but showed poor mechanical properties. Cross-
linking of zein by hexamethylene diisocyanate significantly increased the tensile
strength of the zein electrospun nanofibre mats. The mechanical properties of zein
nanofibres can be improved by combining electrospinning and cross-linking. Zein
can be further processed into resins and other bioplastic polymers, which can be
extruded or rolled into a variety of plastic products.112 With increasing envir-
onmental concerns about synthetic coatings and the ever increasing price of
hydrocarbon-based petrochemicals, there is a great potential for utilizing zein as a
raw material for a variety of non-toxic and renewable polymer applications,
particularly in the paper industry.113,114 Other reasons for the renewed interest in
zein include concern about the landfill costs of plastics, and the increased demand
by consumers for natural substances.
Acknowledgements
The financial assistance from CSIR to NS is acknowledged.
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CHAPTER 11
Silk Fibre Composites

PANYA WONGPANIT,a
ORATHAI PORNSUNTHORNTAWEEb
AND RATANA RUJIRAVANIT*b,c
a
Faculty of Agricultural Product Innovation and Technology,
Srinakharinwirot University, Bangkok 10110, Thailand; b The Petroleum and
Petrochemical College, Chulalongkorn University, Bangkok 10330, Thailand;
c
Center for Petroleum, Petrochemicals, and Advanced Materials,
Chulalongkorn University, Bangkok 10330, Thailand
11.1 Introduction to Silk Fibres and their

Protein Components
Natural fibres can be divided into two main categories based on their origins:
(i) plant-based lignocellulose fibres, which predominantly contain cellulose,
hemicellulose, and lignin; and (ii) animal-based protein fibres, including silk
and wool.1 Humans have utilized natural fibres as textiles for several thousand
years; however, a recent increase in environmental concerns has led to the use
of natural fibres, which are renewable resources, for the development of
environmentally friendly materials, especially in the form of composites.
Among both types of natural fibres, the fabrication of silk-based composites is
of great interest. In this chapter, the structures and important characteristics of
silk fibres and their protein components are briefly discussed, followed by an
overview of the fabrication of silk-based materials, both in the form of raw silk
and regenerated silk. Finally, the preparation of silk-based composites and
their potential applications are intensively reviewed.

219
220 Chapter 11
11.1.1 Silk Fibres and their Sources
Silk fibres are produced by arthropods, especially by members of the class
Arachnida, including mites and more than 30 000 species of spiders, and those
of the class Insecta, which are the larvae of some Lepidoptera, such as butterflies
and moths.2–4 These animals produce the silk fibres for various purposes. For
example, silkworm moths produce the silk fibres to provide shelters in the form
of cocoons during their metamorphosis, while spiders use the silk fibres in the
form of aerial nets to capture prey, to provide structural support in the form of
egg sag suspension threads, to aid dispersal of neonate spiderlings in the wind,
and to escape from predators.2–5
In silk production, proteinaceous materials are first produced by either special
glands, which are generally modified salivary glands in the larvae of some Lepi-
doptera, or by other structures in some immatures and adults of mites and spiders.3
The silk proteins are initially stored in those structures as a liquid before being spun
into fibres during secretion. Although there are many types of silk in nature, each
species of silk producer, except spiders, produce only one silk type.3,6 In the case of
spiders, they can produce at least seven different silk types, including major
ampullate or dragline silk, minor ampullate silk, flagelliform silk, pyriform silk,
aggregate silk, cylindriform or tubuliform silk, and aciniform silk.5–7
Among various kinds of silk producers, silkworms and spiders are intensively
studied for their silk production. Utilization of silk fibres by mankind has been
done at least since 2600 BC by harvesting Bombyx mori silkworms, a member of
the family Bombycidae in the order Lepidoptera, on mulberry trees.3 Up to now,
the mulberry B. mori silk fibres are of the most commercial importance,
especially in the textile industry, due to their large-scale production and unique
luster appearances.5,8,9 Smaller quantities of silk fibres can also be produced by
other species fed on a variety of their natural hosts, such as the productions of
tasar silk, muga silk, and eri silk by silkworms in the family Saturniidae.3 Silk
production by silkworms is also known as sericulture.3,8 For spider silks, there
has been an attempt to use them as either fishing nets or wound closures for
thousands of years.3,6 However, in contrast to silkworms, it is impossible to
harvest spiders for large-scale silk production because of the territorial and
cannibalistic nature of most spiders.3,6 As a result, the utilization of spider silks
is now restricted, although much effort is used to enable their industrialization,
especially by biotechnological production.6 Based on their widespread use, this
chapter will mainly focus on the silk fibres produced by B. mori silkworms. For
spider silks and other silk types, a number of comprehensive reviews have
already been published.2,3,5–7,9
11.1.2 Physical Structure and Chemical Composition

of Silk Fibres
Silk fibres produced by B. mori and other silkworms possess a ‘‘sheath-around-
two-cores’’ structure, containing two protein monofilaments with triangular
sections, named brins, embedded in a sericin layer, which is a water-soluble
protein responsible for the gum-like sticky coating.7,8,10,11 The brins contain
Silk Fibre Composites 221
several macrofibrils, which are bundles of nanofibrils made of fibroin fila-
ments.7 The silk fibres are composed of fibroin proteins in the range of
70–80 wt% and sericin proteins in the range of 20–30 wt%, depending on their
origin and culture conditions.10,12,13 Other minor components, which can be
exclusively found in the sericin layers, include waxes and fats (1.5 wt%), car-
otenoids and minerals (1 wt%), and small amounts of polysaccharides, known
as glycosylated sericin fractions.10
For the chemical composition, the major amino acids that can be found in the
fibroin proteins of the silk fibres produced by B. mori silkworms are glycine
(43%), alanine (30%), serine (12%), tyrosine (5%), and valine (3%).14,15
Therefore, the silk fibroin predominantly contains hydrophobic uncharged
amino acid residues. The fibroin consists of three components: two protein
chains, named as a heavy chain (H-chain, with molecular weight of about 350
kDa) and a light chain (L-chain, with molecular weight of about 25 kDa), which
are linked together via disulfide bonds, and a glycoprotein, named P25, with
molecular weight of about 25 kDa, that is associated via non-covalently hydro-
phobic interactions.16–18 The molar ratio of H-chain:L-chain:P25 is 6:6:1.16–18
The H-chain is more hydrophobic and is responsible for the formation of crys-
talline structures, while the L-chain is more hydrophilic and relatively elastic.16–18
The fibroin possesses three possible conformations, including random-coil,
a-helix (silk I), and b-sheet (silk II).19–21 The first conformation is an amorphous
state, while the a-helix and b-sheet are crystalline structures of the fibroin. The
random-coil conformation and a-helical structure can easily convert to the
b-sheet structure by mechanical agitation or by exposure to heat or to
hydrophilic organic solvents, like methanol.2,14,19–21 The b-sheet structure is
thermodynamically stable and is water-insoluble due to the presence of extensive
inter- and intra-chain hydrogen bonding as well as hydrophobic interactions.14
In general, the fibroin proteins of the silk fibres produced by B. mori silkworms
contain a-helical and b-sheet structures of about 13% and 56%, respectively.22
Thus, the fraction of crystalline structures of the silk fibroin is more than 60%.15
In the case of the sericin, it contains at least six proteins with molecular weights in
the range of 20–400 kDa.10,23 The silk sericin predominantly contains nearly 40%
serine, with smaller amounts of other amino acid components, such as glycine,
aspartic acid, glutamic acid, threonine, and tyrosine.12,24,25 The hydrogen
bonding between hydroxyl groups of these amino acids are responsible for the
glue-like properties of the sericin layer.10 However, unlike the silk fibroin, sericin
possesses only two conformations: random coil (about 63%) and b-sheet (about
35%).24 A low degree of crystallinity and a large content of strongly polar side
group-containing amino acid components make the sericin coating of the silk
fibres become water-soluble.10,12,25
11.1.3 Characteristics of Silk Fibres and their

Protein Components
The silk fibres exhibit outstanding mechanical properties in terms of toughness,
tensile strength, and extensibility (or strain at break) because of the extensive
222 Chapter 11
hydrogen bonding and hydrophobic interactions of their protein components,
as well as a high degree of crystallinity of the silk fibroin.7,8,10 For the B. mori
silk fibres, their average toughness, tensile stress, tensile modulus, and strain at
break are 70 mJ m3, 700 MPa, 10 GPa, and 20%, respectively.2,7,8,10,13,26 These
mechanical parameters are comparable to those of both other natural fibres
and synthetic fibres, including hemp, jute, ramie, coir, sisal, flax, cotton, glass
fibre, and Kevlar.7,8,13,26 The silk fibres can withstand heat up to nearly 200 1C,
with a glass transition temperature (Tg) of 175 1C, before the side-chain groups
of the amino acid residues and the main chain of the silk molecules begin to
degrade at above 200 1C and 300 1C, respectively.10,19 Moreover, compared to
other protein fibres (wool), silk fibres have lower flammability.10 Silk fibres are
biodegradable at a slow predictable rate and show excellent biocompatibility
after the removal of sericin.7–9,27 The unique mechanical properties, slow bio-
degradabilities, and biocompatabilities of silk fibres are valuable for tissue
engineering applications, which require support and transfer of load from the
scaffolding material to the developing tissue.7 As a proteinaceous material, silk
fibres are amphoteric with an isoelectric point (pI) in the range of 4–5.10,15
Compared to wool, silk fibres are more resistant to alkali but more sensitive to
acid.10 The presence of the side-chain groups of amino acid residues also
provides reactive sites for chemical modification of the silk fibres, such as by
graft copolymerization which can further improve the processabilities of the
silk-based composites (see Section 13.3).
11.2 Fabrication and Potential Applications

of Silk-based Materials
Silk fibres have been used as biomedical sutures for decades and as textiles for
centuries.8 However, the original form of silk is unsuitable for use in some
applications. To expand their potential exploitation, the silk proteins have to be
denatured by dissolving the silk fibres in a solvent system prior to processing
into other alternative forms, such as microtubes, films, spheres, hydrogels, or
three-dimensional porous materials. The fabrication of silk proteins into new
morphologies with desirable shapes and structures is called regeneration.
Normally, the regenerated silk-based materials require post-treatments in order
to induce the recrystallization of the silk proteins.
11.2.1 Raw Silk Threads and Spun Silks

After the mature silkworms completely spin their cocoons, which are made of a
single continuous filament of silk, the cocoons are collected and the larvae are
killed by overheating, either by boiling in hot water or exposure to steam.5,9,10
Since the sericin is water soluble, it can be partially removed at this step. When
the sticky sericin coating dissolves in water, the cohesion between the silk fibres
begins to reduce. Hence, the silk fibres in the outer region of the cocoons
are loosened and can be caught on a turning spool.3 Since a single silk filament
is very fine, it has to be wound together, or reeled, to form a continuous
silk thread.3 After being reeled, the raw silk threads, or silk yarns, are
obtained. Normally, a single silk thread is still too fine and too fragile for
end-use, so about nine threads (each thread containing two silk fibres) are
twisted together in order to obtain a silk thread with enough strength to
handle.9 In general, a good quality cocoon can produce silk fibre of length in
the range of 300–1000 m, and about 1500 cocoons are required to manufacture
a kilogram of silk.3 It should be noted that the raw silk threads are often called
raw silk fibres in most research. Furthermore, not all silk fibres are successfully
wound. The remaining unreeled silk fibres as well as shorter or broken filaments
can be used to produce spun silks.
In the textile industry and in some specific applications (especially in bio-
related fields), the raw silk has to be further subjected to a degumming step. In
biomedical applications, the remaining sericin in silk sutures can cause wound
inflammation and allergic reactions, consequently limiting their clinical use.8,9
The degumming step takes advantage of the solubility of the silk sericin in
boiling aqueous solutions containing different degumming agents, such as
surfactants, alkalis, and organic acids.10 However, aqueous alkaline solutions
are the most widely used to degum the raw silk fibres.7,9,10 The degummed silk
fibres are sometimes called the fibroin fibres because it is considered that the
sericin coating is almost completely removed.
Currently, unlike the fibroin which is the core of the silk fibres, the sericin is a
major waste material in raw silk processing. It is estimated that average cocoon
production is one million tonnes worldwide and raw silk processing from this
amount of cocoons can yield sericin waste as high as 50 000 tonnes.25 Although
the sericin protein is unutilized in the textile industry, extensive research has
been carried out to find possible exploitation in other fields, especially in
pharmacological, cosmetic, biomedical, and biotechnological applications. An
important reason why sericin has gained attention in recent years is that the
discarded sericin in wastewater can cause extreme environmental problems, so
the recovery and recycling of sericin are expected to yield economic and social
benefits.12,25 The potential uses of silk sericin have been intensively discussed in
a number of review articles.12,25 Although the sericin proteins can be prepared
in different forms, such as films, hydrogels, and porous materials, the majority
of the sericin-based material is classified as polymer blends;12,25 only a few are
composite materials. Therefore, this chapter will focus on the fabrication of the
regenerated silk fibroin.
11.2.2 Regenerated Silk-based Materials

Figure 11.1 shows an overview of the fabrication of regenerated silk-based
materials into various forms. The degummed silk fibres are first dissolved in a
solvent system to prepare silk fibroin solution for reprocessing into desired
shapes and structures. However, the silk fibres are insoluble in most
aqueous solutions, including dilute acids and bases, due to the highly ordered
orientations of silk fibroin chains in combination with intermolecular hydrogen
224
Chapter 11
Figure 11.1 An overview of the fabrication of regenerated silk-based materials into various forms.
10,11
bonds and hydrophobic interactions. Therefore, to effectively dissolve
the silk proteins, the compact fibrous structure should be swollen and the
hydrogen bond network as well as the hydrophobic interaction should be
interrupted. Moreover, an ideal solvent should penetrate into the silk fibres
without inducing any undesirable reactions, such as depolymerization and
derivatization.10
Normally, the most common solvents used to dissolve the silk fibres can be
categorized into two main groups: aqueous-based and organic-based solvent
systems. The aqueous-based solvent systems are concentrated aqueous salt
solutions, such as lithium bromide28–33 and a ternary solvent of calcium
chloride/water/ethanol.34–38 After complete dissolution, the metal ions have to
be removed from the as-prepared silk fibroin solution, perhaps by dialysis
against distilled water, because the remaining metal ions can further hinder the
regeneration process of the silk fibroin.9 In general, the fibroin proteins in
aqueous condition should be kept at a low temperature (4 1C) in order to
maintain the solution lifetime before gelation occurs (which can be accelerated
by heat and high fibroin concentration under static conditions). Practically, to
increase the concentration of aqueous silk fibroin solution by water evapora-
tion, the solution is slowly stirred in the temperature range of 50–60 1C for the
disruption of inter- and intramolecular forces of the fibroin proteins without
providing strong shear force.39 The presence of physical shear can consequently
induce b-sheet formation, which is the water-insoluble form of silk fibroin.19,21
For organic-based solvent systems, the silk-based materials (either the
degummed silk fibres or the freeze-dried silk fibroin) are able to be dissolved in
organic acids, such as formic acid,40–42 or fluorinated solvents, like 1,1,1,3,3,3-
hexafluoropropan-2-ol (HFIP),43–46 prior to fabrication into desired forms. It
should be noted that the freeze-dried fibroin materials dissolve in these organic
solvents easier than the degummed silk fibres. Another effective solvent system
for the preparation of the fibroin solution is ionic liquid-based, including
1-butyl-3-methylimidazolium chloride.47
After the degummed silk fibres are completely dissolved in the selected sol-
vent, the insoluble materials are removed from the as-prepared silk fibroin
solution by centrifugation and the clear supernatant is subsequently used to
regenerate the silk fibroin into diverse forms by various processing techniques,
as shown in Figure 11.1. For example, the silk fibroin microtubes can be pre-
pared by the dipping and dehydrating method,48 while the micro- and ultrathin
silk fibroin films are prepared using the solution casting procedure38,49 and
spin-assisted layer-by-layer (LBL) assembly,47 respectively. The silk fibroin
micro- and nanospheres are fabricated by spray drying,50 vesicle templating,51
LBL deposition on soluble templates,52 and precipitation in a water-miscible
organic solvent.52 Regenerated silk fibroin hydrogels can be formed with the
aid of heat and sonication.53 For three-dimensional porous structures of the
regenerated silk fibroin, nonwoven mats are prepared by electrospinning,39,54,55
while sponges, or scaffolding materials, are fabricated with the use of either salt
leaching43–46 or freeze drying.37 Other alternative forms of the regenerated silk
fibroin are microfibres, which can be prepared by wet spinning.56,57
226 Chapter 11
Since the dissolution of the silk proteins is achieved at the expense of their
crystalline structure, the as-prepared regenerated silk-based materials are water
soluble. With the use of an aqueous-based solvent system, the silk fibroin
possesses either the random coil conformation (at low fibroin concentrations)
or the a-helical structure (at high fibroin concentrations and in the absence of
physical shear).19,21 Therefore, after being regenerated, post-treatments are
often required to induce the recrystallization (the formation of the b-sheet
structure) of the silk fibroin. An example of the widely used post-treatments is
methanol treatment. After the post-treatment, the regenerated-silk based
materials become water insoluble because of a change in crystallinity and can
be effectively used as biomaterials in a wide range of applications, such as
controlled-release drug delivery systems and scaffolds in tissue engineering
applications. Interestingly, regenerated silk-based materials are now being used
in clinical trials as an anterior cruciate ligament (ACL) replacement. It is also
expected that such material should achieve medical approval for clinical use in
the near future and will be commercially available in both Europe and the USA
within 2014.9
11.3 Silk-based Composites: Fabrication and

their Potential Applications
Conventionally, composites are defined as materials consisting of two or more
distinct and identifiable components, which subsequently provide desirable
properties that cannot be found in the original materials.58,59 However, this
definition is quite ambiguous because, in the case of a polymer/polymer system,
it also covers copolymers and blends. Hence, composites have recently been
defined as materials that can be prepared by aligning strong and stiff con-
stituents, such as fibres and particulates, in a continuous phase that acts as a
binder.59 In general, a filler phase is responsible for the mechanical strength of
the composites, so it is called a reinforcing phase, while a binder phase is called
a matrix. Materials that can be used as reinforcements include fibres, particles,
microspheres, whiskers, and flakes, while those serving as a composite matrix
are metals, ceramics, and polymers. It should be noted that the fillers can be
incorporated into the matrix not only for providing mechanical strength but
also for specific purposes, such as for providing either thermal stability or
electrical conductivity to the resulting composite.
Since the properties of composite materials are determined by the properties
of their components, by the morphology or shape of the filler phase, and by the
nature of the interface between two or more distinct phases, the composite
performance can be greatly varied by simply altering their components.
However, in some cases, the composite fabrication has to be compromised and
the processabilities of the composites should be taken into account because the
combination of some components can possibly lead to undesirable properties.
Another important aspect is that the interfacial adhesion between two or more
discrete phases can play an important role in the mechanical performance of the
fabricated composites.
Composites can be divided into many categories, based on the morphology
or shape of the incorporated fillers, but two intensively studied composites
are particulate-filled composites, which contain discrete particles as a dis-
continuous filler phase in a continuous matrix phase, and fibre-filled compo-
sites. The fabrication of the particulate-filled composites involves suspension of
rigid particles in liquids or molten polymers, while the fibre-filled composites
can be prepared by several techniques, including impregnation of fibre mats,
lamination of impregnated sheets of oriented fibres or woven fabrics, impreg-
nation of three-dimensional woven fabric, and injection or compression
molding of polymers containing reinforcing fibres.58
Silk materials can be used to fabricate both types of composites, particulate-
filled and fibre-filled. Moreover, in silk-based composites, both silk fibres and
regenerated silk fibroin can serve as either reinforcement or matrix. Different
kinds of fillers can be incorporated into silk materials in order to provide not
only mechanical strength but also new favorable properties, such as anti-
microbial activities, biological activities, optical properties, magnetism, and
electrical conductivity. Industrial scale production of silk and its unique
characteristics, especially its degradability and high biocompatibility, are the
key factors that promote intensive study on the development of silk-based
composites for possible exploitation in various fields, ranging from traditional
clothing to highly innovative devices and biomedical end-uses.
Figure 11.2 shows an overview of the fabrication of silk-based composites.
Among a variety of processing techniques, the most widely used methods for
the fabrication of silk-based composites include injection molding, compression
molding, freeze drying, and salt leaching.
The first two methods (injection and compression molding) frequently
involve molten polymers or heated viscous materials. In a conventional injec-
tion molding, either a pre-blended or a physical mixture of polymer pellets/
granules and reinforcing fibres is fed into the hopper before being conveyed by
the flights of a rotating screw to the heated barrel.60,61 As the screw rotates, the
fed materials are accumulated at the front of the screw. At a specific time
interval, the screw stops rotating and moves forward in order to inject the
heated viscous material under high pressure into the mold cavity. After soli-
dification, the mold is open and the fabricated products are ejected.60,61 In
compression molding, a mixture of either polymer pellets/granules or a resin
dough and reinforcing fibres are generally placed in the fixed bottom mold half
before being compressed at an elevated temperature as the top mold half moves
down, usually with the aid of a hydraulic press. After solidification, the top
mold moves up and the composite product is removed from the bottom mold
with the aid of ejector pins.62 Compared to injection molding, compression
molding shows a number of advantages.62 For example, it produces a lower
amount of waste because of the use of a simply designed mold. Moreover, since
compression molding involves a much low shear rate during the composite
fabrication, no severe degradation is observed in the resulting composites. In
addition, high reinforcing fibre volume fractions can be used to fabricate the
composite materials with compression molding, so the obtained compression
228
Chapter 11
Figure 11.2 An overview of the fabrication of silk-based composites.
62
molded products possess better physical and mechanical properties. The
compression molding is also more suitable for the production of composite
materials with a large surface area.62
Freeze drying and salt leaching techniques are generally used for the fabri-
cation of composite materials with three-dimensional porous structures, and
both of them involve either the dispersion of solid particles or the impregnation
of structured materials in a polymer solution. In the freeze-drying technique, as
an aqueous polymer solution is frozen at a temperature below its freezing point,
ice crystals begin to form; this phenomenon is called nucleation.63 These ice
crystals in the frozen polymer solution are subsequently removed by sublima-
tion, yielding the three-dimensional porous structures. Typically, porosity, pore
size, and pore structure of the freeze-dried materials depend on several factors,
such as freezing temperature, freezing rate, and polymer concentration. Nor-
mally, the lower the freezing temperature, the smaller the obtained pore size.
Another important aspect is that the pore shape can be affected by heat transfer
during freezing, because uniform heat dissipation in all directions causes uni-
form ice crystal formation in the frozen polymer solution.63 In the case of salt
leaching, salt granules are used as a porogen instead of ice crystals in the freeze
drying. A desired amount of salt is added to the polymer solution before the
solvent is evaporated, resulting in the embedment of salt granules in a solid
polymeric material. After that, the salt granules, a water-soluble porogen, are
leached out with an excessive wash with water. Thus, the pore size and shape of
the final product are dependent on the size and shape of salt granules. Com-
pared to the freeze-drying technique, the salt leaching has some limitation due
to the leaching step, especially when small molecules, like bioactive compounds
or drug molecules, have to be retained in the final products. However, no
expensive instrument is required in the salt leaching technique.
11.3.1 Silk Fibres as Composite Reinforcement

Recent emphasis on environmental awareness has led to the consideration of
natural fibres as potential alternatives to synthetic fibres for being used as
composite reinforcement. Among two types of natural fibres, namely plant-
based lignocellulose fibres and animal-based protein fibres, the silk fibres
possess several interesting characteristics, such as mechanical properties, ther-
mal stability, biodegradability, and high biocompatibility. As a result, the silk
fibres are used to reinforce various kinds of matrices, including synthetic
polymers, biopolymers, polymer blends, and inorganic matrices. For the silk
fibre-reinforced polymer matrices, the fabricated composites can be called
biocomposites, which are defined as composite materials consisting of natural
fibres as reinforcing elements and biodegradable or non-biodegradable poly-
mers as matrices.13,64,65 The reinforcing silk fibres can be either the raw silk
fibres, which are partially degummed during fibre processing, or the degummed
silk fibres, which are sometimes called the fibroin fibres. The incorporation of
silk fibres in the composite matrices can be in the form of a complex structure,
like knitted silks or woven fabrics. Moreover, in some researches, waste silk
230 Chapter 11
fibres obtained from the textile industry are also used as composite reinforce-
ment. The silk fibres can even be combined with other fibres, e.g. plant-based
fibres or synthetic fibres, before being used as reinforcement because it is
believed that a combination of two or more different types of fibres in a matrix
offers a wide range of properties that cannot be obtained from a single kind of
fibre reinforcement. Since short fibres can be aligned in the composite matrix
with a higher degree of orientation, leading to greater composite performance,
short silk fibre-reinforced composites are extensively studied. Regardless of the
fibre length, the silk fibre content is another key factor affecting the composite
performance. The silk fibre-reinforced composites are good candidates to be
potentially used in diverse fields, from general-purpose plastics to scaffolding
materials in tissue engineering applications. Table 11.1 shows examples of
silk-based composites with silk fibres as composite reinforcement in various
matrices, their improved properties, and their proposed applications.
11.3.1.1 Silk Fibre-reinforced Synthetic Polymers

Waste silk fibres obtained from cocoon reeling, which are undesirable in the
textile industry, can be chopped prior to incorporation into nitrile rubber.66 To
improve the adhesion between the chopped silk fibres and the rubber matrix, a
‘‘resorcinol/hexamethylenetetramine/silica’’ bonding system was used. All
components were mixed by a conventional laboratory open mill before being
compression molded. It was reported that the composite performance was
strongly dependent on the fibre content. An increase in the silk fibre content
increased hardness, tensile strength, tear strength, modulus, heat build-up,
abrasion loss, restriction to solvent swelling, and processability of the fabricated
composite, but simultaneously decreased resilience and elongation at break.
After ageing at 100 1C for 48 h, the silk fibre-reinforced nitrile rubber composite
was able to retain its mechanical strength in terms of tensile strength, tear
strength, and elongation at break. In contrast, the nitrile rubber without silk fibre
reinforcement deteriorated and its strength was expected to reduce after ageing.
Chopped natural silk fibres were used to reinforce poly(butylene succinate)
(PBS), a biodegradable, thermoplastic, and aliphatic polyester.65 The bio-
composite was prepared using the compression molding method and its
mechanical and thermal properties were investigated. The chopped raw silk
fibres were found to markedly improve tensile and flexural properties and
thermomechanical stability of the PBS, while the thermal stability of the silk
fibre/PBS biocomposite was intermediate between the reinforcement and the
matrix. Therefore, the natural silk fibre was an effective reinforcement and
might be a potential candidate for enhancing the performance of biodegradable
polymer matrix resins. These findings are also consistent with the work of
Han et al., who reported that the use of chopped waste silk fibres, which are
industrially available, as reinforcement improved both static and dynamic
mechanical properties of the PBS matrix.67 Moreover, above the glass transi-
tion temperature, the storage modulus of the composite was significantly higher
than that of the PBS matrix.
Table 11.1 Examples of silk-based composite with silk fibres as composite
reinforcement.
Silk-based Fabrication Proposed
compositea method Improved properties applications Ref.
Silk fibre-reinforced synthetic polymers
Silk/PP Compression Mechanical properties, Semi-structural 1

molding degradabilities and structural
applications
Silk/nitrile rubber Compression Mechanical properties – 66
molding
Silk/PBS Compression Mechanical properties, Automobile 67,68
molding thermal properties, applications
water absorption
abilities,
degradabilities
Silk/PLA Extrusion and Mechanical properties, Biomedical and 69–71
injection thermal properties, bioengineering
molding dimensional stabilities, applications,
water absorption environmental
abilities, engineering
degradabilities applications
Silk/PCL Melt mixing Mechanical properties Biomedical 72–76
materials
CNTs-coated silk/ Compression Mechanical properties Commodity 77
PBSA molding materials
Silk/epoxy resin Microbond Interfacial strength Tough composites 78
method
Hand lay-up Interfacial strength, Applications in 79,80
mechanical properties, chemical
chemical resistance surroundings
Silk/PMMA, silk/ Hand lay-up Mechanical properties, Applications in 81–83
PC, silk/ABS chemical resistance chemical
surrounding
Silk/sisal fibres/ Hand lay-up Mechanical properties, Materials for 84–87
unsaturated chemical resistance making chemical
polyester resin, storage tanks
silk/coir fibres/
unsaturated
polyester resin,
silk/glass fibres/
epoxy resin
Silk fibre-reinforced biopolymers
Silk/collagen Freeze drying Mechanical properties, Ligament and 88,89

biocompatibilities tendon tissue
engineering
Silk/collagen/ Freeze drying Biocompatibilities Ligament and 90
hyaluronic acid tendon tissue
engineering
Silk/gelatin Compression Mechanical properties, Single-used 91
molding degradabilities operating
theater trays
232 Chapter 11
Table 11.1 (Continued )
Silk-based Fabrication Proposed
compositea method Improved properties applications Ref.
Silk fibre-reinforced polymer blends
Silk/PP/NR Extrusion and Mechanical properties, Semi-structural 92

compression degradabilities and structural
molding applications
Silk fibre-reinforced inorganic matrices
Silk/MPAHC Sol/gel method Mechanical properties Biocements for 93

ostepedic, ortho-
paedic, or dental
surgery
Silk/PVOH/MMT Freeze drying Mechanical properties Environmentally 94
benign materials
a
PP ¼ polypropylene; PBS ¼ poly(butylene succinate); PLA ¼ poly(lactic acid); PCL ¼ poly(e-
caprolactone); CNTs ¼ carbon nanotubes; PBSA ¼ poly[(butylene succinate-co-(butylene adi-
pate)]; PMMA ¼ poly(methyl methacrylate); PC ¼ polycarbonate; ABS ¼ acrylonitrile/butadiene/
styrene; NR ¼ natural rubber; MPAHC ¼ magnesium-bearing phosphoaluminate/hydroxyapatite;
PVOH ¼ poly(vinyl alcohol); MMT ¼ montmorillonite.
A subsequent study further investigated the long-term stability of the silk

fibre/PBS biocomposite in a wet environment in a comparison to that of the
PBS reinforced with chopped henequen fibres, lignocellulose-based fibres.68
Both PBS biocomposites were fabricated using the compression molding
technique and the hygrothermal effects on silk fibre- or henequen-reinforced
PBS biocomposites were observed at 60 1C and 85% relative humidity (RH). At
the exposure duration of 1000 h, it was found that different degradabilities
of these two reinforcing fibres led to different mechanical performances of
the biocomposites. Although the incorporation of the silk fibres in the PBS
enhanced the water absorption of the biocomposite and consequently caused
a higher hydrolysis degradation of the PBS matrix, the silk fibre/PBS bio-
composite still showed superior mechanical properties because the reinforcing
silk fibres did not degrade. On the contrary, although the henequen/PBS bio-
composite exhibited much lower water absorption, the reinforcing henequen
fibres degraded rapidly and the mechanical properties of the reinforced bio-
composite could not be maintained. As a result, the origin of the natural fibres
significantly affected the biocomposite degradability in the long-term dur-
ability, so the selection of natural fibres as reinforcements must be carefully
carried out and depends on the intended biocomposite applications. These
findings are in accordance with another study which revealed that the origin of
the natural fibres not only affected the degradability of the fabricated com-
posite but also influenced its mechanical properties in terms of bending
strength, bending modulus, tensile strength, tensile modulus, and impact
strength.1 In that work, either silk fibres or jute fibres, lignocellulose-based
fibres, were used to reinforce polypropylene (PP). Compared to the neat PP, the
incorporation of either silk fibres or jute fibres into the PP matrix remarkably
improved its mechanical properties. However, when the two composites were
compared, after 12 weeks of the soil degradation test the silk fibre/PP com-
posite still possessed its excellent mechanical properties while the jute fibre/PP
composite degraded much faster. Again, the results showed that the silk fibre-
reinforced composite retained its mechanical performance longer than the
lignocelluloses-based fibre-reinforced composite because of its slower
degradation.
Natural silkworm silk fibres were gently cut into short fibre fractions before
being used as a reinforcement of poly(lactic acid) (PLA), a synthetic biode-
gradable polymer which is widely used as an implant in medical applications, to
prepare a biodegradable biocomposite.69 The silk fibre/PLA composite was
fabricated using extrusion and injection molding techniques and its mechanical
and thermal properties were studied. It was observed that the silk fibres were
well bonded with the polymer matrix. Compared to the pure PLA, the
mechanical properties, in terms of elastic modulus and ductility, and thermal
stability of the biocomposite were substantially improved. To evaluate the
potential use of the silk fibre/PLA biocomposite in tissue engineering appli-
cations, its biodegradability was subsequently studied in vitro at specific time
periods.70 In that study, the reinforcement of silk fibres increased both stiffness
and ductility of the PLA, and a biodegradation rate of the silk fibre/PLA
composite was faster than that of the pure PLA, indicating that the incor-
poration of silk fibres not only improved the mechanical properties of the PLA
but also altered its biodegradability.
Regardless of the tissue engineering applications, the silk fibre/PLA bio-
composite is potentially useful as a high-performance material for environ-
mental engineering applications.71 Compared to pure PLA, although the
incorporation of the silk fibres in the PLA matrix reduced the thermal stability
of the fabricated composite, the stiffness at high temperature (in the range of
70–160 1C) and dimension stability were markedly enhanced. The biodegrad-
ability of the biocomposite was carried out with the use of a well-known pro-
tein-digesting enzyme: proteinase K. The silk fibres increased the water
absorption of the biocomposite, so the silk fibre reinforcement enhanced
the hydrolysis of the PLA matrix. Moreover, the enzymatic degradation of the
reinforcing fibres also promoted the biocomposite degradability. Since the
degradation of the silk fibre/PLA biocomposite increased with increasing the
silk fibre content, it was suggested that the degradability of the PLA matrix
should be controlled by altering the amount of the silk fibres in the fabricated
composite.
The degummed silk fibres were used to reinforce a film of poly(e-capro-
lactone) (PCL), a biodegradable polyester.72–76 The silk fibre-reinforced PCL
composite was prepared by melt mixing. The influence of the reinforcing silk
fibres on the mechanical properties, viscoelastic properties, and crystallization
behavior of the resulting biocomposites was investigated. The results showed
that the incorporation of the silk fibres into the PCL matrix improved the
mechanical properties of the biocomposites in terms of tensile and flexural
234 Chapter 11
strengths. Moreover, both the storage modulus and loss modulus also increased
with increasing fibre content. The silk fibres improved the crystallization, both
isothermal and non-isothermal processes, of the PCL matrix. Since the silk
fibre-reinforced PCL biocomposite has a potential use as a biomedical material,
the effect of electron beam irradiation (widely used sterilization in the medical
industry) on the properties of the biocomposite was further studied. It was
observed that the irradiation induced both the cross-linking and the degrada-
tion of the polymer components and the mechanical properties of the bio-
composite were directly dependent on the irradiation dose. The tensile and
flexural strengths moderately increased with increasing irradiation dose before
dramatically decreasing as the irradiation dose increased above 150 kGy. The
irradiation was also found to reduce the overall crystallization rate of the PCL
matrix due to the cross-linking. This negative effect became more dominant in
the non-isothermal crystallization, compared to isothermal crystallization.
Biodegradable composites containing poly[(butylene succinate)-co-(butylene
adipate)] (PBSA), an aliphatic polyester, as a polymer matrix and silk fibres
coated with carbon nanotubes (CNTs) as a reinforcement were prepared by
the compression molding method.77 Compared to the uncoated silk fibres, the
CNT coating provided better interfacial interaction between the matrix and the
reinforcement, leading to greater mechanical performance of the resulting
CNT-coated silk fibre/PBSA composite, which should be beneficial for use as a
commodity material, such as a general-purpose plastic.
A degummed silk fibre/epoxy composite was prepared and the interaction
between the silk fibre and the resin matrix was studied.78 With the use of the
microbond test, the mean interfacial shear strength was found to be about
15 MPa, indicating that the silk fibre efficiently reinforced the epoxy resin, one
of the most important thermosetting polymers. Although epoxy resin is widely
used as either a structural material or a structural adhesive, it is brittle and has
poor resistance to crack propagation. Since the silk fibres possesses high tensile
strength and high extensibility, comparable to those of glass fibres or synthetic
polymers, the determined interfacial shear strength suggests another exploita-
tion of the silk fibre in tough composites.
Not only silk fibres but also waste silk fabric obtained from the textile
industry has been used as composite reinforcement.79–83 The waste silk fabric-
reinforced epoxy resin was prepared by a hand lay-up method. The incor-
poration of the waste silk fabric into the epoxy resin increased the mechanical
performance of the fabricated composite in terms of tensile, flexural, impact,
and compression strength. The waste silk fabric-reinforced epoxy resin showed
good chemical resistance to acids, alkalis, water, and organic solvents. More-
over, good interfacial adhesion between the reinforcing silk fabric and the
epoxy resin also resulted in a lower void content of the fabricated composite.
However, it was suggested that interfacial bonding should be improved by
surface treatment of the silk fabric, probably with the use of an appropriate
coupling agent.
In later work,81–83 the influence of tougheners and waste silk fabric content
on mechanical properties and chemical resistance of the fabricated composites
were studied. In the presence of toughener, the epoxy resin was prior mixed
with three thermoplastics, including poly(methyl methacrylate) (PMMA),81
polycarbonate (PC),81 and an acrylonitrile/butadiene/styrene (ABS) copoly-
mer,82,83 before being reinforced with the waste silk fabric. Both in the absence
and presence of toughener, the mechanical performance of the composites in
terms of tensile, flexural, and impact properties were found to increase with
increasing in the fabric content. For their resistance to moisture, the water
absorption capacity of the silk fabric-reinforced epoxy resin decreased in the
presence of toughener but increased with an increase in the fabric content. In
addition, the toughened epoxy resin composites also displayed good chemical
resistance to acids, alkalis, and organic solvents, indicating good efficacy for
applications in chemical surroundings.
Recently, it has been reported that either the silk fibres or the silk fabric can
be combined with other types of reinforcing fibres, including sisal fibres,84,85
coir fibres,86 and glass fibres,87 prior to use as reinforcement of resin matrices,
either unsaturated polyester resin84–86 or epoxy resin.87 The composites were
prepared with the use of the hand lay-up technique. Again, the fabricated
composites exhibited an increase in mechanical performance in terms of tensile,
flexural, and compressive properties, and with good chemical resistance, indi-
cating their potential use for making chemical storage tanks. It was also
reported that the interfacial adhesion between the natural fibres and the resin
matrix was able to be improved by treating the fibre surface with an alkaline
solution.84–86
In conclusion, although the silk fibres are effectively used as reinforcement
for various kinds of synthetic polymer matrices, the silk fibre content in the
polymer matrix should be optimized in order to obtain the desired composite
performance. Another important aspect is that the hydrophilic nature of the
silk fibres can enhance the water absorption of the fabricated composite and
subsequently promote hydrolysis of the biodegradable synthetic polymer
matrix. Compared to plant-based fibres, the silk fibres (animal-based fibres) are
better composite reinforcements from the viewpoint of long-term stability
because of their slower biodegradability.
11.3.1.2 Silk Fibre-reinforced Biopolymers

Since both silk fibres and some biopolymers possess excellent biocompat-
ibilities, most silk fibre-reinforced biopolymers are prepared for possible use in
the biomedical field, particularly for tissue engineering applications. However,
a slow in vivo degradation of the silk fibres makes the fabricated composites
valuable for longer regenerating engineered tissues, such as bones, cartilages,
ligaments, and tendons.
The incorporation of knitted silk fibres in a collagen matrix has been
reported.88,89 The raw silk fibres were used to prepare a warp knitted scaffold
with the use of a knitting machine. After being degummed, the knitted silk was
immersed in an acidic collagen (type I) solution and then freeze dried. The
fabricated knitted silk/collagen composite scaffold exhibited optimal internal
236 Chapter 11
space, good biocompatibility, and good mechanical strength. The composite
scaffold not only increased structural and functional ligament repair, but also
enhanced the differentiation of human embryonic stem cell-derived mesench-
ymal stem cells (hESC-MSCs), the most widely used stem cells in tendon tissue
engineering. Thus, the knitted silk/collagen composite scaffold should be a
promising material for practical utilizations in both ligament and tendon tissue
engineering.
In another study, the knitted silk-based composite scaffold was prepared by
immersing the knitted silk in an acidic mixture solution of collagen and hya-
luronic acid, or hyaluronan, one of the glycosaminoglycans.90 The composite
scaffold was then fabricated by the freeze-drying technique. The biocompat-
ibility of the composite scaffold was evaluated in vitro by assessing anterior
cruciate ligament (ACL) and T-lymphocyte cultures. The utilization of the
composite scaffold as artificial ligaments in ACL reconstruction and its influ-
ence on angiogenesis was investigated in vivo. The initial attachment and
proliferation of human ACL cells on the composite scaffold were higher than
those on the knitted silk; however, the in vitro immune responses between the
two scaffolds were not different. After being grafted in the knee joint of a dog
for 6 weeks, both granulation tissue and new blood vessels were observed in the
dog grafted with the composite scaffold, while implantation of the knitted silk
scaffold did not provide the formation of reparative tissues. These findings
indicated that the knitted silk scaffold with a lyophilized collagen/hyaloronic
acid mixture was biocompatible in vitro and promoted the formation of new
blood vessels as well as the cell migration in vivo, suggesting its potential for
ligament and tendon tissue engineering.
A natural silk fibre-reinforced gelatin composite was fabricated in order to
prepare a biodegradable composite to serve as a single-used operation theater
(OT) tray because traditional OT trays, which are made of steel, require labor-
and time-consuming processes for cleaning and sterilization after operations.91
The composite was fabricated by undirectionally arranging three layers of silk
fibres among four layers of gelatin film before being compression molded.
Compared to the pure gelatin matrix, the silk fibre-reinforced gelatin composite
showed higher tensile strength, tensile modulus, bending strength, bending
modulus, and impact strength. The fabricated composite rapidly degraded after
being buried in compost soil by losing more than 50% of its original weight
within 24 h.
11.3.1.3 Silk Fibre-reinforced Polymer Blends

Short silk fibres were used to reinforce a thermoplastic elastomer blend of
polyethylene (PE) and natural rubber (NR).95 To obtain a uniform dispersion
of silk fibres in the PE/NR blend, the silk fibres were first added to the molten
thermoplastic before blending with the NR. With a proper mixing sequence, it
was observed that each silk fibre was coated with a thermoplastic layer. The
‘‘resorcinol/hexamethylenetetramine/silica’’ bonding system was also used to
promote the adhesion between the reinforcing fibres and the matrix. Although
the bonding agent was able to improve composite performance, a longer curing
time was required for full adhesion development. An increase in the silk fibre
content substantially increased both the tensile and the tear properties of the
fabricated composite; however, its elongation sharply reduced. Further exam-
ination revealed that the silk fibre acted as a mechanical anchor between the
thermoplastic phase and the NR. Nevertheless, the effect of the silk fibre
content on the mechanical properties of the composite became less dominant at
high thermoplastic blend ratios.
A recent study reported on the preparation of a silk fibre-reinforced PP/NR
blend.92 The NR lumps were cut into small pieces and blended with PP in an
extruder before being cold pressed. The obtained PP/NR blend film was used to
fabricate a silk fibre-reinforced composite by arranging four layers of the blend
films among three layers of the silk fibres before being compression molded.
The silk fibre-reinforced PP/NR blend exhibited good degradability, but the
NR in the blend dramatically decreased the mechanical properties of the
fabricated composite in terms of tensile strength, tensile modulus, bending
strength, bending modulus, and hardness (except impact strength). Hence,
g-radiation was employed to improve the mechanical properties of the com-
posite. It was found that the mechanical properties of the composite increased
with increasing radiation dose to a certain limit before starting to decrease. At
low radiation doses, the radiation not only removed moisture from the
composite but also induced cross-linking between the silk fibre and the blend,
leading to better fibre/matrix adhesion and an increase in mechanical properties
of the composite. In contrast, high radiation doses possibly induced polymer
chain scissions of silk fibre, PP, and NR, hence consequently reducing com-
posite performance. The silk fibre-reinforced PP/NR composite was degradable
so it was an environmentally friendly material which should be suitable for
semi-structural as well as structural applications.
11.3.1.4 Silk Fibre-reinforced Inorganic Matrices

Raw silk fibres were used as reinforcement for a composite cement of magne-
sium-bearing phosphoaluminate/hydroxyapatite (MPAHC).93 The silk fibres
were treated with nitric acid and subsequently soaked in an aqueous ethanol
solution. After drying, the treated silk fibres were mixed with the cement before
molding. Among the test specimens, the composite cement containing 0.5 wt%
of the treated silk fibres possessed the highest splitting strength and Vickers
micro-hardness. The incorporation of treated silk fibres in the cement enhanced
the mechanical strength of the studied system while maintaining its bio-
compatibility. As a result, MPAHC reinforced with the treated silk fibres
should be a preferential candidate for biocement purposes, which might find
applications in the fields of osteopedic, orthopedic, or dental surgery.
The silk fibres and a number of biologically based fibres, such as hemp,
ramie, and bamboo top, were used as reinforcements of clay aerogel to fab-
ricate environmentally benign materials for a wide range of end-uses.94
Poly(vinyl alcohol) (PVOH) was also incorporated in the clay aerogel to
238 Chapter 11
improve its mechanical performance. An aqueous PVOH solution was mixed
with a suspension of sodium montmorillonite (Na1-MMT) before adding
chopped fibres to the PVOH/clay mixture at various fibre contents. The fibre-
reinforced PVOH/clay composites were prepared via the freeze-drying proce-
dure. The chopped fibres formed a woven-like structure in the clay aerogel.
The mechanical properties in terms of compressive strength and moduli of the
fabricated composites depended on the type of reinforcing fibres, but increased
remarkably with increasing the fibre content.
11.3.2 Silk Fibres as a Composite Matrix

The unique fibril structure, the good mechanical properties, and the presence of
amino acid components containing strongly charged and reactive side groups
make silk fibres an attractive substrate for the embedment of different kinds of
inorganic nanoparticles. The embedded nanoparticles provide new biological
activities and physical properties to the silk fibres, such as antimicrobial
activities, optical properties, magnetism, and electrical conductivity, hence
further expanding their possible use as smart textiles in various fields. Table
11.2 lists examples of silk-based composites with silk fibres as a composite
matrix containing various types of fillers, their improved properties, and their
proposed applications.
Cubic silver chloride nanocrystals were synthesized on silk fibres via sequential
dipping of the silk fibres in alternating solutions of silver nitrate and sodium
chloride.96 It was proposed that the negative charge at the surface of the silk
fibres provided the immobilization of Ag1 prior to reaction with Cl, leading to
the formation of an initiator seed of AgCl, and the sequential dipping further
resulted in the growth of nanocrystals. The AgCl-incorporated silk fibres should
be used as either a photocatalyst in water-splitting applications or as an anti-
bacterial agent. Another attempt to develop silk nanocomposites with anti-
bacterial properties as potential textiles for hygienic clothing, wound healing, and
Table 11.2 Examples of silk-based composite with silk fibres as composite

matrix.
Silk-based Fabrication Improved
composite method properties Proposed applications Ref.
AgCl/silk Sequential Optical properties, Photocatalyst in water- 96
dipping antimicrobial splitting applications,
activities antibacterial materials
AgBr/silk Sequential Antimicrobial Hygienic clothing, wound 97
dipping activities healing, and medical
applications
Ag/silk Biotemplate Optical properties Chemical sensing, separation, 98,99
redox catalysis, data storage, and
photonic materials
CdS/silk Bio-inspired Optical properties Compartment in nanodevices 100
process for bio-related fields
medical applications was the incorporation of silver bromide nanoparticles in silk
yarns via sequential dipping in alternating solutions of silver nitrate and potas-
sium bromide under ultrasound irradiation.97 Compared to the traditional
synthesis procedure, the sonochemical method yielded smaller AgBr nano-
particles with narrower particle size distribution on the silk yarns.
Silver nanoparticles were successfully incorporated into degummed silk fibres
using a biotemplate redox procedure at room temperature, a green and mild
technique to fabricate functional hybrid nanocomposites for possible utiliza-
tion in the fields of chemical sensing, separation, catalysis, data storage, and
photonic materials.98,99 The tyrosine in the silk fibroin was found to mediate
the reduction of Ag(I) ions to Ag(0) under alkaline conditions, resulting in the
in situ formation of Ag nanoparticles in the silk fibroin fibres, an inorganic/
organic hybrid nanocomposite.
Regardless of the nanoparticles of Ag and its complexes, there is a report on
the synthesis of cadmium sulfide (CdS) strings and hexagons on the degummed
silk fibres through a room temperature bio-inspired process.100 Unlike the Ag
which provided an antibacterial effect, the CdS, a typical semiconductor, gave
optical properties to the fabricated composites. In that study,100 the silk fibres
which acted as reactive substrates provided in situ formation sites of CdS
nanocrystals and simultaneously controlled the arrangement of the resulting
nanocrystals into specific patterns. The CdS/silk nanocomposite displayed
photoluminescence, suggesting its utilization as a compartment in nanodevices
for bio-related fields.
Although most studies focused on the use of the silkworm silk fibres as a
composite matrix, the preparation of spider silk fibre composite was also
reported.101,102 Dragline spider silks from Nephila edulis were impregnated with
different kinds of inorganic nanoparticles, such as superparamagnetic magne-
tite (Fe3O4), gold, and CdS, by immersing the spider silk fibres in the as-pre-
pared nanocolloidal sol.101,102 Compared to bare dragline spider silk fibres, the
impregnation of inorganic nanoparticles showed a negligible effect on the
mechanical properties of the resulting composite fibres in terms of tensile
strength, elongation at break, and tensile modulus. However, the combination
of the natural strength and elasticity of the spider silk with physical properties
of the added inorganic nanoparticles (including magnetism, electrical con-
ductivity, and semiconductivity) should be useful as smart structural fabrics in
a wide range of applications.
11.3.3 Regenerated Silk as a Composite Reinforcement

Several research studies aimed to use the regenerated silk fibroin either in the
form of microspheres or microparticles as reinforcement for various kinds of
biodegradable synthetic polymers in order to develop new composites with
desirable properties, such as biocompatibility, biodegradability, and envir-
onmentally friendly characteristics, for specific purposes. Since silk fibroin
powder is already commercially available, the widespread utilization of the
regenerated silk as a composite reinforcement is now being promoted. Table 11.3
240 Chapter 11
Table 11.3 Examples of silk-based composites with regenerated silk as com-
posite reinforcement.
Silk-based Fabrication Improved Proposed
compositea method properties applications Ref.
Silk fibroin/PLA Supercritical Foam cell size, Bioplastic foams 103
CO2 foam cell density
Freeze drying Biocompatibilities Hepatic tissue 104,105
engineering
applications
Silk fibroin/poly- Solution Mechanical – 106
(e-caprolactone- casting properties
co-D,L-lactide)
Silk fibroin/WPU Solution Mechanical – 107
casting properties
Silk fibroin/chicken Melt Thermal Biodegradable 108
egg shells/SEBS processing properties applications
a
PLA ¼ poly(lactic acid); WPU ¼ waterborne polyurethane; SEBS ¼ poly(styrene-b-ethylene/
butylene-b-styrene).
lists some examples of silk-based composites with regenerated silk as composite

reinforcement in various matrices, their improved properties, and their proposed
applications.
Microcellular biodegradable silk fibroin/PLA composite foam was fabricated
using supercritical carbon dioxide to develop a new bioplastic.103 The silk
fibroin powders were mixed with a PLA solution of various powder content
prior to foaming. Compared to the neat PLA foam, the composite foam pos-
sessed a larger cell size and a higher cell density. In addition, both the cell size
and the cell density increased with increasing powder content. Regardless of the
silk fibroin powder content, the cell size and the cell density of the composite
foam were also affected by the saturation temperature and pressure during the
fabrication. Both the cell size and the cell density were simultaneously reduced
with an increase in the saturation temperature and pressure. Therefore, the
microcellular structure of the composite was able to be controlled by altering
either the silk fibroin powder content or the processing conditions.
A composite scaffold composed of silk fibroin, either in the form of micro-
spheres or microparticles, and a PLA matrix was prepared using the freeze-
drying technique.104,105 The hepatocellular compatibility and the inflammatory
response of the silk fibroin/PLA composite scaffold were subsequently eval-
uated with the use of human hepatocellular liver carcinoma cells (HepG2) and
mouse RAW264.7 macrophage cell lines, respectively. The scaffold possessed
interconnected pore structures and the silk fibroin uniformly dispersed in the
PLA. The HepG2 cells attached, spread, and proliferated on the composite
scaffold much better than those on the PLA scaffold, indicating that the
embedded silk fibroin improved the cell–sponge interactions. The inflammatory
response of the macrophages to the composite scaffold was also found to be
lower than that to the PLA scaffold. Thus, the silk fibroin/PLA scaffold was a
good candidate for hepatic tissue engineering due to its higher hepatocellular
compatibility with lower inflammatory response.
The silk fibroin particles were also incorporated into poly(e-caprolactone-
co-D,L-lactide), which is a biodegradable polymer.106 The composite film was
fabricated using the solution casting procedure and its thermal and mechanical
properties were investigated. The silk fibroin particles were homogeneously
dispersed in the polymer matrix. Although the decomposition temperature and
the degradation rate of the fabricated composite decreased with an increase in
the silk fibroin content, its mechanical properties (such as storage modulus and
hardness) were improved.
Waterborne polyurethane (WPU) reinforced with silk fibroin powders has
been reported.107 To achieve a good dispersion of the silk fibroin powder in the
WPU matrix, both the negative charges in the periphery and the small particle
size were key factors. It was found that the dispersion of the silk fibroin
powders in the WPU matrix was strongly dependent on the powder content. At
high silk fibroin powder content, the silk fibroin/WPU dispersion exhibited
lower negative zeta potentials and larger particle sizes, leading to poorer
mechanical properties of the resulting composite film. On the other hand, at
lower powder content, the added silk fibroin dispersed well in the WPU matrix
and the silk fibroin/WPU composite film showed a remarkable increase in
mechanical properties in terms of tensile strength and modulus.
In a recent study, silk fibroin powders and chicken eggshells were simulta-
neously used as bio-fillers to reinforce poly(styrene-b-ethylene/butylene-b-
styrene) (SEBS), a member of the styrene tri-block copolymers, in order to
prepare a composite for biodegradable applications.108 The fibroin/eggshell/
SEBS composite was fabricated via a melt processing method with the use of a
co-rotating twin-screw extruder. The fibroin played a key role in improving the
interfacial interaction between the reinforcements and the polymer matrix.
Compared to the pure SEBS, the thermal stability of the resulting composite
was found to increase.
11.3.4 Regenerated Silk as a Composite Matrix

Traditionally, when the regenerated silk fibroin was used as a matrix, the
composites were prepared using two main procedures. In the first method, the
silk proteins were processed into the desired morphology before the fillers/
reinforcements were impregnated into the as-prepared regenerated fibroin.
Another preparation technique involved mixing of the filler dispersion with
the silk fibroin solution prior to fabricating the composite material into the
favorable form. However, a spin-assisted layer-by-layer assembly has recently
been proposed as a novel route to prepare regenerated silk-based nano-
composite films.
Since the regenerated silk fibroin is one of the most promising biomaterials
for bone tissue engineering applications because of its appropriate degrad-
ability, several research studies have tried to provide an osteogenic environment
by the incorporation of biominerals that can be found in native bone.
242 Chapter 11
In addition, although the regenerated silk fibroin has several favorable
biological characteristics, including excellent biocompatibility with low
inflammatory and immunological response, the important drawbacks of the
silk proteins are poor mechanical properties and low dimensional stability,
especially for highly porous structures. As a result, much effort has been
expended to improve its physical performance by the fabrication of composite
materials with the use of organic nanofillers (whiskers obtained from bio-
polymers). Table 11.4 shows some examples of silk-based composites with
regenerated silk as a composite matrix containing various types of fillers, their
improved properties, and their proposed applications.
11.3.4.1 Regenerated Silk with Inorganic Fillers

Titanium dioxide (TiO2) nanoparticles were embedded in a silk fibroin matrix
with the use of either the sol/gel technique109,110 or the freeze-drying method.111
The TiO2 nanoparticles dispersed well in the fibroin matrix without reagglo-
meration and it was also reported that the embedment of TiO2 was able to
induce the structural transition of silk fibroin from silk I to silk II. Compared to
the fibroin film, the TiO2/fibroin nanocomposite showed higher mechanical and
thermal properties but lower water solubility. However, the excessive TiO2
nanoparticles tended to disrupt the crystal structure in the nanocomposite.
Since the fabrication of the nanocomposite via these techniques avoided the
reagglomeration of the TiO2 nanoparticles, it was proposed that the fabricated
TiO2/fibroin nanocomposite should find potential applications in the biome-
dical field.
Hydroxyapatite (HA), an important constituent component of bone, was
incorporated into silk fibroin materials, including nonwoven net, sheet, film,
and scaffold, with the use of various techniques, such as a wet-mechan-
ochemical route,112,113 a co-precipitation method,114–119 an alternating lami-
nation,120 and an alternate soaking method.121–123 Moreover, a thin film of
HA/silk fibroin was successfully prepared by using matrix-assisted pulsed laser
evaporation (MAPLE).124 Since the presence of the calcium ion plays a key role
in the formation of HA crystals on polymeric surfaces, the HA/silk fibroin
composite was fabricated by simply soaking the fibroin film containing calcium
chloride in simulated body fluid (SBF).125
It was reported that the thermal stabilities of the HA/silk fibroin composites
were improved while their microstructures and tensile properties were
dependent on the HA content in the composite.117,126 Since HA is a brittle
compound, an excess HA content can presumably make the resulting compo-
site become more brittle. Another possible reason why the silk fibroin com-
posite at high HA content exhibited a reduction of tensile properties was
inorganic/organic phase separation due to low interfacial adhesion. As a result,
a number of research studies focused on the enhancement of the interfacial
adhesion between the HA and the fibroin. For example, the silk fibroin was
pretreated with different kinds of chemical reagents, including an aqueous
solution of inorganic salts,127 an aqueous alkali solution,128–130 and the enzyme
Table 11.4 Examples of silk-based composite with regenerated silk as com-
posite matrix.
Silk-based Fabrication Improved Proposed
compositea method a properties applications Ref.
Regenerated silk with inorganic fillers
TiO2/silk Sol/gel method, Mechanical Biomedical 109–111

fibroin freeze drying properties, applications
thermal properties
HA/silk Wet-mechan- Mechanical Bone tissue 112–143
fibroin ochemical route, properties, engineering
co-precipitation, thermal applications
lamination, properties,
alternative soaking, osteoconductive
MAPLE, soaking properties
in SBF
Gelation Enhanced wound Wound dressing 144
healing properties materials
Silica/silk Protein fusion, Osteoconductive Bone tissue 145–147
fibroin deposition properties engineering
applications
Silicon/silk Co-precipitation Induction of HA Bone tissue 148
fibroin formation engineering
applications
CaSiO3/silk Freeze drying Mechanical Tissue engineering 149
fibroin properties, water applications
absorption
abilities, induction
of HA formation
MMT/silk Solution Mechanical Environmentally 150
fibroin intercalation properties friendly materials
from tissue engi-
neering scaffolds
to biodegradable
structural
materials
LBL assembly Mechanical Biomedical 151
properties applications
Ag/silk LBL assembly Mechanical Photonic and 152
fibroin properties, optical optical
properties utilizations
Regenerated silk with organic fillers
Chitin Freeze drying Dimensional stabi- Tissue 37

whiskers/ lities, mechanical engineering
silk fibroin properties applications
Cellulose Solution casting Mechanical Biomedical 152
whiskers/ properties applications
silk fibroin
Alginate beads/ Porogen leaching, Sustained drug Drug delivery 153,154
silk fibroin, freeze drying release applications
PLGA/
silk fibroin
a
HA ¼ hydroxyapatite; MAPLE ¼ matrix-assisted pulsed laser evaporation; SBF ¼ simulated body
fluid; MMT ¼ montmorillonite; LBL ¼ layer-by-layer; PLGA ¼ poly(lactide-co-glycolide).
244 Chapter 11
130,131
proteinase K, to disentangle surface fibrils for an effective contact between
the inorganic and the organic phases. The structural modification was also done
to improve the interaction between the HA and the fibroin by grafting the silk
fibroin with other polymers, namely poly(g-methacryloxypropyltrimethox-
ysilane) [poly(MPTS)],132–134 poly{2-[O-(1 0 -methylpropylideneamino)carbox-
yamino]ethyl methacrylate},135 and poly(4-methacyloyloxyethyl trimellitate
anhydride) [poly(4-META)].136 An alternative strategy to improve the inter-
facial adhesion was to use a blend of silk fibroin and various kinds of polymers,
such as hyaluronic acid137 and chitosan,138 a deacetylation derivative of chitin,
as an organic matrix for the HA. The HA/fibroin composites showed potential
use as either bone in growth or implant fixation in bone tissue engineering
because a number of cells, including human bone mesenchymal stromal
cells (BMSCs),119 osteoblasts (MC3T3-E1),120 human bone marrow stem cells
(hMSCs),121 osteosarcoma SaOs2 cells,124 and rat marrow mesenchymal cells
(MMCs),134 were able to attach, spread, and differentiate well on the composite
surfaces, indicating the osteoconductivity. To promote the osteogenesis, the
HA/silk fibroin composites were combined with bone morphogenetic protein-2
(BMP-2).121,139 Another study reported that the HA/silk fibroin composite
scaffold also promoted the healing of mandibular bone defects in canines.140 It is
important to note that although B. mori silk fibroin was widely used as a matrix
for the HA composites, the HA was successfully embedded in the film of
genetically engineered Nephila clavipes spider silk.141 In addition, the HA was
not only deposited on regenerated silk material but also on silk fabric, or silk
cloth, by using either co-precipitation or soaking in order to prepare bioactive
materials for orthopedic implants.142,143 It was further reported that the
deposition of the HA was able to be promoted by the immobilization of urease
onto the surface of silk cloth.143
Although most research has mainly focused on the osteoconductive prop-
erties of the HA/silk fibroin composite, one study showed that the addition of
the HA to the silk fibroin gel enhanced epidermal recovery from full thickness
porcine skin wounds.144 The HA/fibroin composite promoted the wound
healing process, re-epithelization, and matrix formation, indicating its potential
use as effective wound dressing material.
Regardless of the HA, the incorporation of silica particles into either the silk
fibroin film or the spider silk film also yields osteoinductive properties.145–147
Furthermore, the growth of HA in the HA/silk fibroin composite was found to
be accelerated in the presence of silicon, one of the components in bone,
although the morphology of the resulting HA crystal depended on the silicon
content.148
A composite scaffold of silk fibroin with bioactive wollastonite (CaSiO3), a
classic example of calcium silicate-based ceramics, was fabricated via freeze
drying.149 Compared to the neat silk fibroin scaffold, the composite scaffold
showed superior properties in terms of compressive strength, compressive
modulus, surface hydrophilicity, and water-uptake capacity. The wollastonite/
fibroin composite scaffold possessed good bioactivity by inducing HA forma-
tion on its surface after being soaked in the SBF. The composite scaffold was
compatible with cell line L929 of mouse connective tissue, a fibroblast-like cell,
indicating its utilization in tissue engineering applications.
MMT clay was used as nanoscale reinforcement for the fabrication of MMT/
silk fibroin nanocomposite via the solution-intercalation technique.150 An
aqueous fibroin solution was added to a clay suspension before being lyophi-
lized to obtain a nanocomposite powder. It was reported that the clay dis-
persion in the fabricated nanocomposite was dependent on the solution pH.
The MMT/silk fibroin nanocomposite prepared under acidic conditions, or at a
pH below the pI of fibroin protein, exhibited better clay dispersion compared to
that prepared at neutral pH or at a pH above the pI, due to a stronger clay–silk
fibroin interaction. Regardless of the clay dispersion, the solution pH also
influenced the conformation of the fibroin protein. Under acidic conditions, the
incorporated clay acted as an efficient nucleator of b-sheet formation of silk
fibroin. On the other hand, the silk fibroin in the nanocomposite prepared at
neutral pH preserved the random coil conformation. Since both the clay dis-
persion and the fibroin conformation play an important role in the composite
performance, the solution pH should be properly adjusted. It was further
suggested that the MMT/silk fibroin nanocomposite should be an envir-
onmentally friendly material which was suitable for a large number of uses,
ranging from a tissue engineering scaffold to biodegradable structural
materials.
A recent study reported on the preparation of MMT/silk fibroin nano-
composite film by using the spin-assisted LBL assembly.151 Compared to the pure
fibroin, the composite film was highly transparent and possessed superior
mechanical properties in terms of strength, modulus, and toughness. As the
MMT was replaced by a Langmuir monolayer of densely packed silver, the
resulting composite film was found to be highly reflective, or mirror-like,
although the silver nanoplates improved the mechanical properties in the same
trend as the MMT. It was also proposed that this LBL strategy should be a new
route for the preparation of nanocomposite films with exceptional mechanical
properties for biomedical applications, and that a mirror-like silk-based nano-
composite should possibly be beneficial for both photonic and optical
utilizations.
11.3.4.2 Regenerated Silk with Organic Fillers

A silk fibroin film was reinforced with cellulose whiskers in order to improve its
mechanical performance.152 The composite film was fabricated by using the
solution casting technique and its mechanical properties were determined in terms
of tensile strength, ultimate strain, and tensile modulus. Compared to pure fibroin
film and pure cellulose film, the cellulose whisker-reinforced fibroin film possessed
much higher mechanical properties. It was also reported that a highly ordered
structure of cellulose whiskers was able to induce the structural transition of silk
fibroin from random coil to b-sheet structure at the whisker/matrix interface.
In the case of highly porous materials like sponges or scaffolds, the induction
of b-sheet formation of the regenerated silk fibroin in order to prepare
246 Chapter 11
water-insoluble materials usually causes high shrinkage, so it is difficult to
control their size and shape. To improve the dimensional stability of the silk
fibroin sponge, there was an attempt to utilize chitin whiskers as a nanofiller.37
The chitin whisker-reinforced silk fibroin nanocomposite sponge was fabri-
cated by adding a fibroin solution to a chitin whisker suspension with
mechanical stirring before being freeze dried. The fabricated nanocomposite
sponge was then treated with an aqueous methanol solution to induce b-sheet
formation of the fibroin. The chitin whiskers homogeneously embedded in the
fibroin matrix and the incorporation of the chitin whiskers successfully reduced
shrinkage of the nanocomposite sponge after methanol treatment. It was also
reported that the added whiskers not only improved the dimensional stability
of the nanocomposite sponge but also increased its compression strength. To
further evaluate the feasibility of the nanocomposite sponge for tissue engi-
neering applications, the cytocompatibility test was done against the L929 cell
line. The test cell attached well to the surface of the nanocomposite sponge, and
the incorporation of the chitin whiskers into the fibroin matrix significantly
promoted cell spreading. As a result, the fabricated nanocomposite sponge was
not cytotoxic and should have potential as a scaffolding material.
Various kinds of microparticles, including alginate beads and poly(lactide-
co-glycolide) (PLGA), were embedded in silk fibroin scaffolds via either the
porogen leaching or the freeze-drying method, not for reinforcement but for
control-release purposes.153,154 The microparticles were loaded with desired
substances, such as insulin-like growth factor I (IGF-I), bovine serum albumin
(BSA), or insulin, prior to the embedment. Compared to the free micro-
particles, those embedded in the fibroin scaffold provided a superior sustained
release rate, indicating their possible use as effective vehicles for drug delivery
applications.
11.3.5 Miscellaneous
13.3.5.1 Silk-reinforced Silk Composites
Silk-based composites are being developed to fabricate new types of bio-
composites with unique characteristics. For example, all-silk-reinforced silk
composites have good mechanical properties because of the high interfacial
adhesion between the reinforcement and the matrix. Moreover, the high bio-
compatibility also makes these silk-based composites one of the most promising
materials for biomedical applications.
An electrospun-knitted silk composite scaffold for ligament tissue engineer-
ing has been reported.155 The raw silk fibres were fabricated into a knitted
scaffold before being subjected to the degumming process. After that, silk
fibroin nanofibres were electrospun onto the degummed knitted silk scaffold to
cover its macropores. With the use of an inclined electrospinning technique, the
electrospun silk fibroin nanofibres adhered better on the knitted silk scaffold.
The potential of the electrospun-knitted silk composite for ligament tissue
engineering applications was examined in a comparison with a PLGA scaffold
(one of the leading biomaterials used as tissue engineered ligament replace-
ment). The cell culture test was performed against fibroblast cells for 28 days
and it was found that the fabricated silk composite showed a higher cell
adhesion with good cell proliferation and higher collagen content but possessed
a lower in vivo degradability.
A uniaxially aligned silk fibre was used to reinforce the silk fibroin matrix in
order to prepare a fibre-reinforced composite with good interfacial adhesion.156
The silk fibre was degummed and treated with an aqueous LiBr solution before
being impregnated in the silk fibroin solution. The composite film was then
fabricated using the solution casting technique. The LiBr pretreatment was
found to improve the impregnation of the silk fibre in the fibroin matrix by
creating loosened fibroin segments on the silk fibre surface which were able to
interact with the fibroin matrix and acted as mechanical interlocking between
the reinforcement and the polymer matrix. Thus, the interfacial adhesion was
increased and the composite performance, in terms of mechanical and thermal
properties, was consequently enhanced. It was suggested that the high-perfor-
mance all-silk composite material is a promising candidate for various
applications.
A silk fibroin sponge was reinforced with silk fibroin microparticles to fab-
ricate a protein/protein composite scaffold with desirable mechanical properties
for in vitro osteogenic tissue formation.157,158 The composite scaffold was
prepared by using the salt leaching technique and it was reported that the silk
microparticles dramatically increased the compressive modulus of the fabri-
cated scaffold. An increase in the scaffold stiffness better mimicked the
mechanical properties of native bone and enhanced the osteogenic differ-
entiation of hMSCs, indicating the feasibility of the composite scaffold for bone
repair.
11.3.5.2 Silk-based Conductive Composites

A recent emergence of interactive and smart textiles has led to the development
of silk-based conductive composites. Either conductive polymers or CNTs can
be incorporated into the silk material in order to provide the electrical con-
ductivity. The fabricated silk-based composites should be beneficial for a wide
range of applications, from traditional clothing to highly innovative electronics
and biomedical end-uses.
A number of electrical conductive polymers, including polypyrrole (PPy),
polyaniline (PANI), and poly(3,4-ethylenedioxythiophene) (PEDOT), were
used to coat silk materials, such as nonwoven web, fibres, and fabric, via in situ
polymerization in order to fabricate conductive polymer/silk composites.159–161
The conductive polymers entirely covered the surface of the silk materials
without damaging their physical morphology, indicating the high affinity
between conductive polymers and silk. The conductive polymer coating did not
affect the mechanical properties of the silk materials, but remarkably increased
their thermal stabilities and provided electrical properties. The application of
248 Chapter 11
the conductive polymer/silk composites should not be limited to only the
clothing industry but should also include the fields of technical textiles, from
interactive and smart textiles to bio-based conductive composites for biome-
dical applications.
Regardless of the conductive polymer coating, the silk-based conductive
composites were fabricated by the incorporation of CNTs, either single-walled
carbon nanotubes (SWNTs) or multi-walled carbon nanotubes (MWNTs), in
silk fibroin nanofibres via the electrospinning technique.162–164 The CNTs were
successfully embedded and well-aligned in the fibroin nanofibres, resulting in
drastically increased mechanical strength and electrical conductivity. There-
fore, the electrospun nanocomposite showed good efficacy in bone tissue
engineering in which load-bearing tissue requires electrical conductivity for cell
growth.165
11.3.5.3 Silk-based Composites as Biosensors

In most research, the regenerated silk fibroin is used as an important compo-
nent in biosensors to provide a biocompatible environment for the enzyme
immobilization. Regardless of the regenerated silk, the biosensors also consist
of inorganic components, such as metal nanoparticles and CNTs.
A sensitive and stable amperometric biosensor was developed via the
immobilization of acetylcholinasterase (AChE) on a platinum electrode mod-
ified with gold nanoparticles and silk fibroin for the determination of carba-
mate and organophosphate pesticides, such as carbofuran, phoxim, and methyl
paraoxon.166 A colloidal mixture of gold nanoparticles and silk fibroin was
used as a composite matrix for the immobilization of AChE, an important
enzyme for the proper functioning of the central nervous system in humans
which is quite sensitive to pesticides,167 prior to deposition on the platinum
electrode. It was reported that the gold nanoparticles improved the sensitivity
and response of the biosensor while the fibroin not only stabilized the immo-
bilized AChE but also prevented its leakiness from the platinum electrode. The
fabricated biosensor showed good sensitivity to carbofuran, phoxim, and
methyl paraoxon, with good reproducibility and acceptable stability, indicating
its efficacy as a promising tool for pesticide analysis.
In another study,168 a colloidal mixture of gold nanoparticles and silk fibroin
was also used as a composite matrix for enzyme immobilization to fabricate a
third-generation biosensor on the basis of direct electrochemistry of redox
enzymes for hydrogen peroxide detection. Horseradish peroxidase (HRP), a
heme-containing enzyme, was immobilized in the composite matrix before
being used to modify a glassy carbon electrode (GCE). The gold/silk fibroin
composite matrix was found to effectively maintain the bioactivity of the
immobilized HRP. The fabricated biosensor exhibited fast amperometric
response, low detection limit, and wide linear range to H2O2, with good
reproducibility and long-term stability, suggesting its potential utility as an
electrochemical biosensor for H2O2 detection.
The preparation of amperometric biosensors based on immobilization of
tyrosinase on a MWNTs/cobalt phthalocyanine (CoPc)/silk fibroin composite
modified GCE for determination of 2,2 0 -bis(4-hydroxyphenyl)propane
(bisphenol A, BPA), a key component in both PC and epoxy resin which has
been identified as a harmful pollutant, has been reported.169 A suspension of
MWNTs functionalized with CoPc was used to modify the surface of the GCE.
After drying, a mixed solution of silk fibroin and tyrosinase was dropped on the
surface of the MWNTs/CoPc/GCE in order to prepare the biosensor. The silk
fibroin maintained the bioactivity of the tyrosinase while the MWNTs and the
CoPc gave excellent inherent conductivity and good electrocatalytic activity to
the composite, respectively.
Instead of enzyme immobilization, a subsequent study reported another
potential procedure to fabricate an electrochemical BPA biosensor based on
dendrimer immobilization.170 It is known that dendrimers are highly branched
and monodisperse macromolecules with a globular structure containing many
terminal functional groups and cavities for hosting other molecules.171 In that
previous work,170 poly(amidoamine) (PAMAM) was immobilized on a gold/
silk fibroin/GC electrode and the results showed that the modified electrode
displayed better electrochemical response to BPA, with a higher adsorption
capacity of BPA, compared to the unimmobilized GCE. Regardless of its high
sensitivity, the biosensor also possessed good reproducibility and acceptable
stability, indicating its possible use for tracing BPA amounts either in plastic
products or in the environment.
11.4 Conclusions
Although the silk-based composites can be prepared in a diversity of forms,
including fibres, films, and sponges or scaffolds, with several interesting prop-
erties, most research still focuses on the utilization of the silk fibres produced by
mulberry silkworm moths due to their large-scale production. Advances in
biotechnology perhaps can enhance silk fibre production from alternative
sources, such as non-mulberry silks, spider silks, and recombinant silks, thus
resulting in further development of the silk-based composites with desirable
properties. It is also believed that the silk-based composites are good candidates
for replacing synthetic materials as well as conventional composites in the near
future because of their environmentally friendly characteristics. In addition, the
excellent biocompatibilities of the silk-based composites might lead to their
end-uses in high value-added applications, especially in bio-related fields.
Hence, silk-based composites can be considered as promising materials for both
environmental engineering and bioengineering applications.
Acknowledgements
The authors would like to acknowledge the Ratchadapiseksomphot Endow-
ment Fund, Chulalongkorn University, for providing a postdoctoral fellowship
to O.P.
250 Chapter 11
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CHAPTER 12
Hybrid Composite Structures

from Collagenous Wastes and
Environmental Friendly
Polymers: Preparation,
Properties and Applications
M. ASHOKKUMAR, P. THANIKAIVELAN* AND
B. CHANDRASEKARAN
Advanced Materials Laboratory, Centre for Leather Apparel & Accessories

Development, Central Leather Research Institute (Council of Scientific &
Industrial Research), Adyar, Chennai 600020, India
12.1 Introduction
Biomass is a non-fossil biological material derived from living organisms such
as plants, animals and microorganisms.1–3 Different sources of biomass include
wood,4 crops,5 animal wastes,6 agriculture wastes,7 food wastes,8 industrial
wastes,9 etc. The effective utilization of biomass or its wastes as valuable hybrid
biocomposite materials has received considerable attention due to environ-
mental concerns and global warming. There is a dire need to source and
develop appropriate technology to satisfy this requirement. Global livestock
animal biomass meant for food, fibre and labor accounts for nearly 200 million

257
258 Chapter 12
tonnes (dry weight), with approximately 3 billion counts. Livestock animals such
as cattle, goats, sheep, pigs, etc., are slaughtered by humans primarily for their
meat. The skins, hides and other organic matter such as bones and offal con-
taining high level of proteins are typically disposed off as wastes from the
slaughterhouse.10 On the other hand, the leather industries utilize a huge amount
of solid proteinaceous biomass wastes, skins and hides, for making leather.
The transformation of skin/hides into leather requires a series of chemical
and mechanical operations and generates solid, liquid and gaseous wastes.11,12
Figure 12.1 shows a schematic of conventional leather processing, associated
solid wastes generation and some of their major applications. The unavoidable
solid wastes generated from the leather industry can be broadly classified
as untanned collagenous, tanned collagen–chromium complexes and non-
proteinous wastes. The preliminary stage in the leather making process gen-
erates an enormous quantity of proteinaceous biomass wastes such as raw
fleshings, hide or skin trimmings, limed split wastes and limed fleshings.13 These
wastes contain high-value proteinaceous material, namely collagen. The other
form of proteinaceous solid wastes predominantly contain heavy metals, such
as trivalent chromium, or vegetable tannins bound with collagen and include
wet blue trimmings, chrome-tanned leather shavings, wet blue split wastes,
vegetable tanned leather shavings, a combination tanned leather shavings,
buffing dust, crust trimmings, finished leather trimmings, leather scrap wastes,
etc. Traditionally, these wastes are either disposed off, dumped or burned in
suburban fields, thereby producing toxic chromium compounds and resulting
in a significant environmental impact as well as destroying an economically
valuable and protein-rich biomass.14–18 The deposition of huge quantities of
tanned and untanned proteinaceous solid wastes in land or by burning in an
open area is a potential danger to public health due to the possibility of oxi-
dation of chromium(III) to toxic chromium(VI).19 Recent environmental reg-
ulations and escalating landfill costs have encouraged both researchers and
industrialists to seek appropriate solutions for the utilization of these wastes.
Presently, the untanned protein wastes are commercially utilized for the pro-
duction of gelatin,20 partially hydrolyzed collagen or glue.21 In some parts of
the world, glue production from animal bones and hides has been a lucrative
business.22 The chromium-containing protein wastes are utilized for making
different low-value products, such as the formation of paste board,23–25
sheets,26–29 insulators,30,31 biogas generation,32 inner soles for shoes, etc.33,34
Among the different technologies employed for the conversion of waste
biomasses, the preparation of hybrid composite materials plays an important
role. Recent advances in composite science and technology and natural fibre
research and development offer significant opportunities for the preparation of
new and improved biocomposite materials from renewable resources that are
optionally recyclable, biocompatible and biodegradable, thereby enhancing
global sustainability. The development of biocomposite materials is consistent
with the principles of green chemistry and engineering, which pertain to the
design, commercialization and use of processes and products that are techni-
cally and economically feasible while minimizing the generation of pollution.
Hybrid Composite Structures from Collagenous Wastes
259
Figure 12.1 Schematic showing the generation of different types of solid wastes from a conventional leather processing chain and their present
utilization areas.
260 Chapter 12
This chapter provides detail on the recent research, development and future
directions of hybrid biocomposites from natural proteinaceous biomass wastes
such as skin trimmings and leather wastes employing environmentally friendly
biopolymers.
12.1.1 Eco-benign Polymers

Environmentally friendly polymers have been defined as polymers that degrade
in the environment by several mechanisms and culminate in complete biode-
gradation so that either no residue remains in the environment or only non-
toxic residues are left. In recent years, there has been growing interest in the use
of biodegradable polymers for different applications in order to reduce different
forms of environmental pollution. Eco-benign polymers and their derivatives
are diverse, abundant and important for life. These polymers are mostly pro-
duced from living organisms or through non-toxic precursors and reactions.
They exhibit fascinating properties and are of increasing importance for dif-
ferent applications. These polymers are broadly classified into two main areas:
renewable and non-renewable polymers. Essentially, renewable biodegradable
polymers utilize a renewable resource (i.e., animal or plant by-products)35 in
the development of the polymer, rather than a non-renewable (i.e., petroleum-
based) resource.36 Obviously, long-term research and development focuses on
the utilization of various renewable/biodegradable polymers for a variety of
applications, including as composites. Some of the environmentally friendly
polymers include cellulose, starch, chitin, chitosan, proteins, peptides, alginate,
gelatin, poly(lactic acid), locust bean gum, pectin, plant exudate gums, poly(3-
hydroxybutyrate), polyamide, etc. Originally, biopolymers were intended to be
used in packaging,37 farming38 and other industries with low strength
requirements. The performance limitations and high cost of biopolymers are
major barriers for their widespread acceptance as substitutes for traditional
non-biodegradable synthetic polymers. The high cost of some eco-benign
polymers compared with traditional polymers is not due to the raw material
costs for biopolymer synthesis but mainly to the low volume of production.
New and advanced synthetic procedures for preparing biopolymers need to be
developed for their high-value applications. The challenge for the development
of biodegradable polymers lies in the fact that they should be stable during
storage or use and again should degrade only when they are disposed of after
their intended lifetime. Eco-benign polymers reinforced with natural biomass
can produce novel biocomposites to replace and substitute different synthetic
polymer fibre-reinforced composite materials for various applications.
12.1.2 Bio-based Materials versus Environment

Interest in eco-friendly or green polymeric materials is growing rapidly owing
to concerns with the toxicity of degraded products and the limited availability
of petroleum-based polymers. Biodegradable polymers from natural or biolo-
gical resources have received much attention due to the fact that they are
Hybrid Composite Structures from Collagenous Wastes 261
biocompatible and biodegradable. When a biodegradable material (neat
polymer, blended product or composite) is obtained completely from renewable
resources, it may be termed as a green polymeric material. The lifecycle of the
renewable polymeric materials is a carbon-neutral process, and their use may
reduce carbon dioxide accumulation in the atmosphere and dependence on
valuable raw materials. Within the past few years, there has been a dramatic
increase in the utilization of environmental wastes, including plant and animal
biomasses. The animal biomass wastes, such as skin, hides, hairs, bones, etc.,
and the plant wastes, such as leaves from flax, jute, hemp, pineapple leaf, sisal,
etc., have been utilized for making new types of environmentally friendly
composite materials.39 However, animal biomass waste, if not properly man-
aged or utilized, can adversely impact the environmental stability by suppres-
sing water quality through surface runoff and erosion, direct discharges to
surface waters, spills and other dry-weather discharges, and leaching into soil
and ground.40 Natural biomass wastes are now emerging as viable alternatives
to form multifunctional composites either alone or in combination with dif-
ferent polymers for various applications in automotive parts, building, struc-
tural and packaging materials. The advantages of using natural biomass waste
are low cost, easy availability, sustainability, recyclability, biodegradability and
waste minimization.41 Another important application for the natural biomass
waste is in the area of biomaterials. A biomaterial intended for biological
applications must possess certain specific characteristics. The most fundamental
requirement is being biocompatible, in other words, not having any adverse effect
on the host. Therefore, in biomedical applications, the traditional composite
structures with a non-biocompatible matrix or reinforcement are being sub-
stituted by engineered biocomposites.42 Hence, it is imperative to consider the
environment and lifecycle aspects of a material, including bio-based materials or
composites, before attempting their development and application.
12.1.3 What are Biocomposites?

Biocomposites are made from a biodegradable polymer as a matrix material
and natural or biodegradable fibres as reinforcing elements. They generally
have low cost, low density, high toughness, significant strength, thermal
properties, enhanced energy recovery and biodegradability. Biocomposites are
capable of undergoing decomposition, primarily through the enzymatic action
of microorganisms, to carbon dioxide, methane, inorganic compounds or
biomass in a specified period of time. Matrix biodegradable polymers may be
obtained from renewable resources, especially from plants and animals, and
have gained much importance over petroleum-based biodegradable polymers
in recent years. Biodegradable reinforcement fibres comprise renewable agri-
cultural, animal and forestry feedstocks (biomass), including wood, agri-
cultural waste, grasses and natural plant fibres composed of carbohydrates
such as sugars, starch, lignin and cellulose,43 as well as animal protein fibres.
Biocomposites prepared by the combination of biomass and biopolymers
provide the necessary performance either alone or in combination with
262 Chapter 12
petroleum-based polymers and offer a sustainable path to achieve eco-friendly
materials. However, the need to produce 100% bio-based composites as sub-
stitutes for synthetic materials is not immediate. Here, different proteinaceous
animal biomass wastes generated from the leather industry can be utilized as a
matrix polymer or reinforcement fibre for making hybrid biocomposites that
can substitute and compete with traditional synthetic polymer-based compo-
sites, thereby reducing the environmental impact and increasing their economic
viability.
12.1.4 Value Added Products from Skin and Leather Wastes:

Current Trends
A great deal of attention from various researchers is being paid to the valor-
ization of chromium-free and chromium-containing proteinaceous wastes from
the leather industry. The chromium-free solid proteinaceous wastes such as skin
trimmings, fleshings and limed trimmings are predominantly used for the
preparation of gelatin,20 poultry feed,44 biodiesel45,46 and fertilizer.47,48 Gelatin
is a proteinaceous material obtained by the partial hydrolysis of collagen
sources. Most of the methods of gelatin manufacture involve the utilization of
bovine hide, cattle bones, pig skin, fish skins, tendons, etc. Hence, skin trim-
ming wastes form an important source for gelatin manufacture. Gelatin pos-
sesses a wide range of applications due to its functional properties such as
biodegradability, biocompatibility and physiological functions.49 Fleshings are
mainly used for vermicomposting to form nutrient-enriched organic manure50
as well as biodiesel production.44,51 Except gelatin, all the other products
obtained from proteinaceous wastes of skins are found to be less proficient,
having low-value applications. Hence, there is a great opportunity to convert
the skin wastes into new innovative products for high-value applications. Along
this line, several initiatives have been made in the recent past. Isolation of
valuable protein products from untanned leather wastes by thermal and
enzymatic treatments has been attempted for food applications.52 Enzyme-
based hydrolysis is becoming popular to extract collagen or its polypeptides
from skin or limed trimmings.44,53–55 Although these initiatives open up new
avenues for the utilization of untanned proteinaceous wastes, there is still
scope for high-value applications such as biomaterials, pharma products, etc.
A major concern is the purity of collagen for biomedical applications; however,
it has been shown that the collagen extracted from bovine hide wastes is pure
and has increased biological and physiological properties.55,56
Another form of leather industry waste, chromium-containing proteinaceous
wastes generated during the process of transformation of hides and skins into
leather, is also being investigated by several groups for their utilization.57,58
Most methods aim at recovery and isolation of the protein fraction, and are
generally based on alkaline (using CaO, MgO or NaOH),59–62 acidic63 or
enzymatic hydrolysis.64,65 The chromium-containing proteinaceous wastes are
also utilized for the formation of composites such as pasteboards,23–25
sheets,26–29 glue,22,66 etc. Among various products reported for the utilization
of chromium-containing proteinaceous wastes, the formation of composite
sheets plays a distinctive role due to their ease of processability and improved
mechanical and thermal properties. Moreover, the developed composite sheets
find wide applications in leather goods, footwear, automotives, furnishings, etc.
In the past, some researchers used chrome-tanned leather shaving wastes to
prepare a powder, after multistep disintegration, which was used as a filler for
butadiene–acrylonitrile rubber.67 Several patents were filed for converting the
leather wastes into semi-continuous sheet or boards for a variety of applica-
tions.23–29 Chrome-tanned collagen wastes were used, beside silica, as a filler for
rubber mixes containing synthetic cis-1,4-polyisoprene rubber and as a dis-
persing agent.68 The polymerization of poly(methyl methacrylate) onto leather
wastes to form composite sheets has been reported and the modification is
claimed to improve the finishing, dyeing and stability of the formed composite
sheets.28,69,70 Composite sheets have also been prepared by the addition of
synthetic polymers like poly(vinyl chloride) (PVC) to leather waste.71,72 Leather
wastes were also used as a filler in thermoplastic polymer composites to increase
their mechanical and thermal properties.73 Chemically modified short leather
fibres were compression molded into a plasticized PVC matrix74, and dioctyl
phthalate plastisols75,76 were further added to form composites with improved
properties. Efforts have been made to form polymer composites by using
chromium-containing wastes with butadiene rubber77 and PVC, which were
found to have improved properties.78 Most of these methods employ expensive
and toxic chemicals and involve noxious reactions, leading to unsatisfactory
composite materials. To overcome these deficiencies, eco-friendly polymers and
materials can be employed to form multifunctional composite sheets from
chromium-containing protein wastes. In this chapter, we have discussed briefly
the development of hybrid composite materials with multifunctional properties
by utilizing collagen-containing protein wastes generated from the leather
industry and environmentally benign polymers.
12.2 Biomaterials from Chromium-free

Proteinaceous Wastes
Collagen, a widely used biopolymer, has been found to possess numerous
biomedical applications. Collagen is extracted from different forms of living
organisms and is classified accordingly in different types. Type I collagen is the
most abundant protein present in animals. It has outstanding mechanical
properties and is present in virtually every extracellular tissue. Some of the
major applications of collagen include wound healing,79 drug delivery, con-
trolled transfer of therapeutic agents,80 substrate culture of living cells,81 gene
delivery82 and replacement/substitutes for human skin,83 owing to its excellent
biocompatibility and biodegradability. Collagen is a natural substrate for cel-
lular attachment, growth, differentiation and promotes cellular prolifera-
tion.84,85 However, the fast biodegrading rate and the low mechanical strength
of the untreated collagen scaffold are crucial problems that limit the further use
264 Chapter 12
of this material. Glutaraldehyde-based cross-linking of the collagen-based scaf-
folds is a widely used method to modify the biodegradation rate and to improve
the mechanical properties.86 Alternatively, different cross-linking agents, such
as formaldehyde, chromium, acyl azide and dimethyl suberimidate,87–89 as
well as synthetic polymers such as polyurethane,90 poly(ethylene oxide)91 and
poly(glycolic acid),92 have been used to obtain optimized hybrid biomaterials
based on collagen. However, these biomaterials have some limitations regarding
their stability, cytotoxicity and degradation properties. The cross-linking agents
or synthetic polymers have an exacerbating effect on the calcification of pros-
thesis materials93 and are also cytotoxic due to post-implantation depolymer-
ization and monomer release.94 To overcome these limitations, it has been shown
that the collagen can be blended with suitable natural or biopolymers instead of
synthetic polymers or cross-linking agents to improve the biomaterial stability
and biological properties. Incorporation of human growth hormone or curcumin
into collagen films has been shown to enhance the wound reduction property and
cell proliferation at the dermal wound site.80 Dermal skin substitutes (mem-
branes) made of collagen and glycosaminoglycan were found to be suitable
substrates for the culture of human epidermal keratinocytes.81 The scaffold
formed by blending collagen with chitosan is a potential candidate for dermal
applications with enhanced biostability and biocompatibility.95 Composite
materials were formed by blending collagen with polycaprolactone96 and
poly(L-lactic acid)97 for cell proliferation and their applications in tissue repair.
The chromium-free proteinaceous wastes, such as skin trimmings, fleshings and
limed trimmings, contain a valuable protein, collagen. There is great potential to
extract collagen from these proteinaceous waste materials and utilize it as a
biomaterial for a number of biomedical applications. This calls for the devel-
opment of hybrid biomaterials for biomedical applications utilizing the collagen
extracted from proteinaceous wastes and natural polymers without employing
toxic cross-linking agents.
12.2.1 Formation of Hybrid Films Using Collagen and

Natural Polymers
The process for the formation of hybrid films is shown schematically in
Figure 12.2. The hide trimming wastes collected from a local tannery at
Chennai were soaked, limed, dehaired, relimed, fleshed and delimed completely
following a conventional leather processing procedure to remove unwanted
proteins and materials.98 The delimed hide pieces were subjected to gradient
dehydration using acetone and methanol to completely remove the moisture.
Finally, the hide pieces were thoroughly dried in a vacuum drier and made into
fine powder using a Wiley mill of mesh size 2 mm. A known weight of hide
powder was dissolved in 0.5 N glacial acetic acid by heating at 45 3 1C to
obtain a known concentration of collagen solution. The formed collagen
solutions (C) were blended with different biopolymers, such as 2-hydroxyethyl
cellulose (HEC),99 starch (ST) or soy protein (SP),100 to obtain viscous
homogeneous solutions. These homogeneous solutions were poured into Petri
dishes and air-dried at room temperature to obtain the desired biopolymer/
collagen hybrid films. All the developed hybrid films had an average thickness
of 30 10 mm.
12.2.2 Characteristics of Collagen/Biopolymer Hybrid Films

The prepared collagen/biopolymer hybrid films have been characterized for
their mechanical, biological and thermal properties. As seen from Figure 12.3a,
the C/HEC hybrid films exhibit a linear increase in the tensile stress as the HEC
concentration increases when compared to collagen/starch/soy composite films.
The percentage elongation or strain of the C/HEC hybrid films decreases
slightly as the concentration of the HEC increases. Similar trends were
observed for C/HEC hybrid films prepared from different concentrations of
collagen, such as 10, 6.7 and 5 mg mL–1.99 The tensile stress values of the C/ST/
SP hybrid films increase with an increase in the concentration of starch (up to
75 wt%) but decrease with an increase in the concentration of soy protein
(see Figure 12.3b). C/ST/SP hybrid films display an inverse relationship
between tensile stress and strain, depending on the type of biopolymer.100
Figure 12.2 Schematic of formation of hybrid biocomposite films.99,100
(a) (b)
Figure 12.3 Stress–strain plots of the developed collagen/biopolymer hybrid films:

(a) C/HEC hybrid films;99 (b) C/ST/SP hybrid films.100
266 Chapter 12
The elongation or strain values of C/ST/SP hybrid films increase with the increase
in the concentration of soy protein and decrease with the increase in the con-
centration of starch. In other words, an increase in the concentration of soy protein
provides more extendibility, while an increase in the concentration of starch pro-
vides more strength in the hybrid films when compared to pure collagen film.
The thermal stability of all the collagen/biopolymer hybrid films significantly
improved as a function of polymer concentration, as evidenced from the
increase in their degradation temperature. The swelling and the in vitro bio-
degradation of the formed hybrid biomaterials reduced significantly as the
concentration of biopolymer (HEC/SP/ST) increased. Further, it has also been
shown using mouse embryofibroblast cells and MTT [3-(4,5-dimethylthiazol-2-
yl)-2,5-diphenyltetrazolium bromide] assay that the addition of biopolymers
(HEC or starch or soy protein) along with collagen assists cell growth and
proliferation.99,100 The morphology of the formed hybrid films as visualized
through scanning electron microscopy is shown in Figure 12.4. The cross-
section of the hybrid films formed using a higher proportion of biopolymer
shows the formation of tiny micro-pores and increased roughness, in com-
parison to the pure collagen film. The presence of pores and rough structure are
expected to assist in the adhesion and growth of the cells, thereby increasing the
cell proliferation and tissue generation.
Hence, the developed hybrid films exhibit superior mechanical, structural
and thermal properties and are also biocompatible without employing toxic
cross-linking agents when compared to the pure collagen films. Thus, the col-
lagen sourced from skin trimming wastes is shown to be pure and not cytotoxic
and hence suitable for various biomedical applications.
12.3 Flexible Composite Sheets from Chromium-

containing Proteinaceous Wastes
Increasing environmental regulations and escalating landfill costs have
forced researchers to look for alternative methodologies for converting
(a) (b) (c)
Figure 12.4 Scanning electron micrographs showing the cross-sectional images

of the developed hybrid films: (a) pure collagen film; (b) 100/100 wt%
C/HEC hybrid film; (c) 100/50/50 wt% C/ST/SP hybrid film.
chromium-containing proteinaceous wastes into useful products. Although
several application avenues have been reported for the valorization of
chromium-containing proteinaceous wastes, the formation of composite sheets
plays a distinctive role due to their easy of processability and improved
mechanical and thermal properties. Numerous articles and patents can be
found on the conversion of these leather wastes into semi-continuous sheets
or boards for a variety of applications. However, previous methods of sheet
formation could not provide sheets with the desired characteristics. Hence,
our group focused on the formation of flexible composite sheets with
multifunctional properties using chrome-tanned leather wastes and a variety
of environmentally friendly polymers such as 2-hydroxyethyl cellulose,101
poly(dimethylsiloxane) (PDMS)102 and poly(vinylpyrrolidone) (PVP).103 All
the chosen environmentally friendly polymers are water soluble and non-toxic.
HEC is a derivative product of cellulose, a natural organic polymer from
plants, widely used in cosmetics, pharmaceutical and household products.104
PDMS is a non-toxic organosilicon compound and its applications range from
cosmetics and medical devices to food additives.105 Similar to PDMS, PVP is
also a non-toxic polymer with applications ranging from cosmetics, medical
devices and food additives to pharmaceutical and personal care products.106
The amount of polymer has been optimized for achieving desired properties on
the derived sheets. The formed composite sheets have been studied for their
structural, physical and thermal properties.
12.3.1 Flexible Composite Sheet Formation

Figure 12.5 shows a schematic for the formation of flexible composite sheets by
utilizing chromium-containing proteinaceous wastes and environmentally
friendly polymers. The chrome-tanned leather wastes were initially ground into a
fine powder of 2 0.5 mm size in a Wiley mill and stored at room temperature.
The powdered chrome-tanned leather wastes were partially hydrolyzed using
dilute sulfuric acid followed by microwave heating to form a homogenous mass.
The homogenous mass was mixed with environmentally friendly polymers such
as HEC,101 PDMS102 or PVP103 in different weight ratios ranging from 0 to
40 wt%, based on the weight of chrome-tanned leather shaving wastes. In the
Figure 12.5 Schematic of flexible composite sheet formation.

268 Chapter 12
case of PVP, pure collagenous paste (C), prepared by dissolving the skin powder
using 0.5 N acetic acid, was used in conjunction with chrome shaving mass (CS)
in a ratio of 1:10 (on raw weight basis) to form a homogenous mixture (CCS).
The pure collagenous paste was used as a binder for the effective binding of PVP
with chrome shaving mass. The corresponding mixtures were heated at 50 2 1C
for 10 min with stirring and transferred to a glass plate and spread uniformly or
hot pressed. The dried composite sheets were removed from the plates and stored
for further analysis.
12.3.2 Properties of the Flexible Composite Sheets

The mechanical properties of all the developed composite sheets show satis-
factory improvement as the composition of the polymer increases, as shown
in Table 12.1. It is seen that the tensile stress values of PVP- and HEC-
incorporated composite sheets increase as the concentration of the PVP or
HEC increases.101,103 On the other hand, PDMS incorporation in the compo-
site sheets leads to a mixed trend, with an initial increase but a sudden
decrease of tensile stress after 20 wt.% polymer concentration.102 In general,
PVP-incorporated sheets were found to have improved mechanical properties
when compared to that of cellulose- or siloxane-incorporated composite sheets.
The order of mechanical properties of the developed composite sheets can be
arranged as a function of the chosen environmentally friendly polymers as
PVP4HEC4PDMS. The greater mechanical property of PVP-incorporated
composite sheets compared to HEC or PDMS may be due to the strong binding
between the collagen–chromium complex and oxygen functionalities of PVP.
Akin to this, the thermal properties of the composite sheets exhibit a similar
performance, as analyzed through thermogravimetric analysis. In general, a
moderate improvement in the thermal properties of the composite sheets has
been observed as the composition of the polymer increases. The softness of all
the composite sheets is in inverse relation to the tensile property of the devel-
oped composite sheets, as evidenced from Table 12.1. Hence, it is possible to
Table 12.1 Tensile and softness properties of the developed composite

sheets.101–103
Composition of Tensile stress (MPa) Softness (mm)
CS/CCS/polymer
composite sheets
(wt%) PVP HEC PDMS PVP HEC PDMS
100/0 7.37 2.2 – 0.52 0.4 2.13 0.2 – 6.5 0.0
100/2.5 9.28 1.8 0.17 0.08 – 3.51 0.1 6.73 0.0 –
100/5.0 17.47 0.7 0.73 0.28 0.63 0.2 1.95 0.1 6.73 0.0 6.5 0.2
100/7.5 18.90 1.2 1.30 0.21 0.93 0.8 1.69 0.0 6.63 0.1 6.5 0.1
100/10 21.31 5.9 2.10 0.30 1.14 0.5 1.52 0.0 5.40 0.2 5.1 0.1
100/20 – 3.14 0.45 1.72 0.9 – 3.80 0.2 4.1 0.0
100/30 – – 0.58 0.01 – – 6.5 0.1
100/40 – – 0.54 0.07 – – 6.5 0.2
develop multifunctional composite sheets with tunable properties from
chromium-containing proteinaceous wastes using environmentally friendly
polymers.
Scanning electron micrographs of the cross-section of fractured surfaces after
tensile strength analysis of selected composite sheets are shown in Figure 12.6.
The CCS/PVP and CS/HEC composite sheets show a reduction of fibre
pull-out (Figure 12.6a–c) or the presence of pores (Figure 12.6d–f) as the
concentration of PVP or HEC increases, indicating better binding and for-
mation of the hybrid composite sheets. The CS/PDMS composite sheets
(a) (b) (c)
(d) (e) (f)
(g) (h) (i)
Figure 12.6 Scanning electron micrographs of the cross-section (d–f) or tensile frac-
tured surfaces (a–c; g–i) of select composite sheets: (a) 100/2.5 wt% CCS/
PVP; (b) 100/7.5 wt% CCS/PVP; (c) 100/10 wt% CCS/PVP; (d) 100/2.5
wt% CS/HEC; (e) 100/7.5 wt% CS/HEC; (f) 100/20 wt% CS/HEC;
(g) 100/5 wt% CS/PDMS; (h) 100/20 wt% CS/PDMS; (i) 100/40 wt%
CS/PDMS.101–103
270 Chapter 12
(Figure 12.6g–h) have increased cracks and roughness on their surface as the
concentration of the PDMS increases up to 20 wt%, which may be due to
strong binding of PDMS with the chrome-tanned leather wastes. On the other
hand, the presence of a very high concentration of PDMS (40 wt%) resulted in
a collapsed surface without any cracks (Figure 12.6i). These results are in
agreement with the mechanical properties and softness data.
12.4 Conclusion
In this chapter, we have discussed a possible solution for the valorization of
different forms of proteinaceous solid biomasses arising from leather industries
as wastes. The chromium-free proteinaceous wastes were blended with different
natural polymers such as cellulose, soy and starch without employing any toxic
cross-linking agents to form hybrid biomaterials. It has been shown that soy
protein provides elastic properties to the hybrid films while the incorporation of
starch provides more strength when compared to pure collagen film. The
developed hybrid biomaterials were found to have promising mechanical,
thermal, degradation, swelling and biocompatibility properties. Moreover, one
of the biopolymers, collagen, is sourced from collagenous wastes and hence the
developed hybrid films have more commercial viability. Chromium-containing
leather wastes were combined with environmentally friendly polymers such as
HEC, PVP and PDMS to form flexible composite sheets by a simple, efficient
and scalable technique. It has been found that the tensile strength of PVP- and
HEC-incorporated composite sheets increases gradually as a function of con-
centration, while the PDMS sheets show a mixed trend. For all the composite
sheets, the softness property had an inverse relation with the tensile strength.
The presence of pores and fibre pull-outs is found to decrease gradually as the
concentration of polymer increases, indicating better binding and sheet for-
mation, as evidenced by SEM analysis. Attempts are also being made to fab-
ricate multifunctional nanobiocomposites from proteinaceous skin/leather
wastes using nanomaterials such as carbon nanotubes, graphitic nanocarbon
materials derived from leather industry wastes and inorganic nanoparticles and
other environmentally friendly polymers for a variety of applications. The
emergence of nanotechnology as well as biotechnology will provide viable and
profitable solutions for the valorization of these protein-rich wastes in the near
future.
Acknowledgement
The authors gratefully thank the Council of Scientific and Industrial Research
(CSIR), India, for providing financial assistance under the YSA project scheme.
The authors also wish to thank Dr A. B. Mandal, Director, Central Leather
Research Institute, India, for his encouragement. The authors also thank
Mr R. Murali and Ms A. Anumary for their valuable help in carrying out some
of the experiments.
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CHAPTER 13
Spider Silk: The Toughest

Natural Polymer
GANGQIN XU,a GUOYANG WILLIAM TOH,a
NING DUa,c AND XIANG YANG LIU*a,b
a
Department of Physics, National University of Singapore, Singapore
117542; b Department of Chemistry, National University of Singapore,
Singapore 117542; c Singapore-MIT Alliance for Research and Technology,
S16-07, 3 Science Drive 2, Singapore 117543
13.1 Introduction to Natural Spider Silk Fibres

In order to catch prey, many spiders possess different silk types to construct
disparately shaped webs, among which orb-webs are the best studied. In gen-
eral, there are four different kinds of silk in the construction of orb-webs:
dragline silk [also termed as major ampullate (MA) spidroin silk], minor
ampullate (MI) spidroin silk, flagelliform silk and piriform silk. Dragline silk is
used in the building of the framework (frame and radii) and lifeline, as shown in
Figure 13.1, which demonstrates a schematic illustration and real photograph
of an established orb-web of the spider Nephila pilipes. MI silk is utilized to
form a temporary auxiliary spiral to stabilize the web structure and to perform
as a template for the succeeding capture spiral, which is formed by flagelliform
silk. The gluey piriform silk is used as ‘‘attachment cement’’ to interconnect the
different structures in an orb-web.1,2 Dragline silk is the most extensively
characterized spider silk among the miscellaneous types.

275
276 Chapter 13
Figure 13.1 Schematic (left) and photograph (right) of the orb-web of the spider
Nephila pilipes.
Spider dragline silk, used both as the web framework and safety line, is one of
the most intriguing natural material marvels, owing to its exceptional strength
and extensibility as well as light weight and biodegradability.3–6 In contrast to
current petrochemical-based synthetic polymers, the spider spins its ultra-
strong, tough and totally recyclable fibres at ambient temperatures, low pres-
sures and with water as solvent.7,8 Spider silk provides a ‘‘green’’ alternative to
high-performance synthetic fibres because of its environmental and human-
friendly nature. Owing to the extraordinary properties of silk fibres, material
scientists have generated increasing interest in designing and developing nature-
inspired synthetic polymers with novel functions, as well as silk-based
biomaterials.
13.2 General Structure of Silks

Spider dragline silk is a semicrystalline polymer, similar to silkworm silk,
consisting of crystalline and non-crystalline (amorphous) regions.9 The anti-
parallel b-pleated sheet crystals, acting as the interlocks, are inter-dispersed in
the amorphous region containing a range of conformations such as ‘‘random
coil’’, b-turns and a-helices or the more compact and left-handed 3(1)-helices10–16
to form a type of molecular network. The primary structures of silk proteins in
both spider and silkworm are composed of repetitive sequences that can be
separated into small blocks.17 The amino acid sequence in the crystalline region
of Bombyx mori silk fibroin is considered to be (GAGAGS)n (G, glycine; A,
alanine; S, serine) and the sequence in the amorphous region contains Tyr-rich
domains.17 On the other hand, N. pilipes spider dragline silk is composed
mainly of two proteins, spidroin I and II (SP I and SP II).18 These proteins can
be divided into block copolymers with GAGA(A)n blocks and non-crystalline
glycine-rich domains.19
Spider Silk: The Toughest Natural Polymer 277
Table 13.1 A comparison between the structures of the spider N. pilipes
dragline silk and the silkworm B. mori cocoon silk.25
Crystallite Inter-crystallite
size (nm) distance (nm)
Content of
Overall intramole-
Sample content of Crystal- cular Meridional Equatorial
name b-sheets (%) linity (%) b-sheets (%) a b c direction direction
Spider 51 (100%) 22 (43%) 29 (57%) 2.1 2.7 6.5 17.8 13.5
dragline
(10 mm s–1)
Silkworm 49 (100%) 40 (82%) 9 (18%) 2.3 4.1 10.3 4.8 7.2

cocoon silk
Two types of b-structures can be identified in silks: the intramolecular b-

sheets (non-crystalline b-sheets)13 and intermolecular b-sheets (b-crystallites).
The b-crystallites (orthogonal unit cell, a ¼ 1.03, b ¼ 0.944, c ¼ 0.695 nm for
the spider Nephila dragline silk,20 and a ¼ 0.938, b ¼ 0.949, c ¼ 0.698 nm for the
silkworm B. mori silk21), constructed by several neighboring silk protein
molecules, crosslink individual silk protein molecules22 so as to form the
molecular network.13 The intramolecular b-sheets, on the other hand, are
merely normal b-sheets in individual silk protein molecules. As shown in Table
13.1, for the dragline filaments of N. pilipes spiders, 57% of the total b-sheets
are in the amorphous region, as characterized by Fourier transform infrared
spectroscopy (FTIR). The remaining 43% of the b-sheets are b-crystal-
linites,13,23–25 as measured by X-ray diffraction (XRD). In contrast, the
amount of intramolecular b-sheets in B. mori silkworm silk is much less. A
comparison of the primary and crystalline structures between silkworm and
spider silk proteins may explain the difference. B. mori silkworm silk proteins
have significantly longer repetitive b-structural motifs than N. pilipes spider
silk proteins. The b-structure sequence is GAGAGSGAAS(GAGAGS)n,
where n ¼ 1–11 for B. mori silkworm silk, and GAGA(A)n, where n ¼ 4–6 for
N. pilipes spider silk. This is in accordance with the larger crystallinity and
crystallite size along the c direction (i.e. backbone direction) given by XRD
measurements (Table 13.1). Although silkworm silk has larger crystallites,
their inter-crystallite distance is much shorter than that of spider dragline silk,
indicating that the crystallites in silkworm silk are packed much closer com-
pared to spider draglines. The larger crystallites along the c direction and the
shorter inter-crystallite distances may provide the clue why the silkworm silk
contains much less intramolecular b-sheets than spider silk. Since protein
chains containing larger portions of regular b-structures [polyA or poly(GA)]
have a higher possibility to interact with each other to form intermolecular b-
crystallites when the adjacent molecular chains are parallel aligned during silk
spinning, this will suppress to a large extent the occurrence of intramolecular
b-sheets.
278 Chapter 13
13.3 The in vivo Formation of Silks
The unique mechanical properties of spider silk threads and an inability to
domesticate spiders have induced numerous attempts to artificially manu-
facture spider dragline silks from recombinant/reconstituted silk protein26–36
for industrial and medical applications.37 However, the mechanical properties
of the native spider silks have not been reproduced with either reconstituted or
recombinant spider silk proteins,31,38 mainly due to the difficulty in duplicating
a spider’s unique protein dope and spinning process in its natural way.
The difference of rheological behavior between native silk dope and repro-
cessed silk protein solution exhibits different structures of the spinning gel
formed in vivo, in contrast to the one formed in vitro.39,40 Holland et al.40 also
showed remarkable similarity between the rheologies for native spider dragline
and silkworm cocoon silk, despite their independent evolution and substantial
differences in protein structure.
Figure 13.2 shows spider silk processing in the spinneret in its natural way.
Spider dragline silk proteins are stored at concentrations up to 50% (w/v)
known as the silk spinning dope, a highly viscous liquid crystalline solution, in
the MA gland until they are processed into fibres. Rheometry enables us to
examine, in vitro, the spinning forces exerted on the dope in vivo along the
spinning duct, thus imparting an analytical window into the silk spinning
process. It also impresses us with the ability of both spider and silkworm to
store silk proteins over long periods and stand by to spin a fibre instantly.
As displayed in Figure 13.2, during the natural spinning process the proteins
move distally through the tapering duct which is divided into three limbs, where
Figure 13.2 Schematic illustration of the spinning process of spider silk in its natural
state. Silk dope is stored in the gland, then moved through the tapering
duct with three limbs, encountering elongational flow, shear force and
changes in their biochemical environment (ion exchange, decrease in pH
and increased concentration). The preliminary fibre is reached after an
internal drawdown process in the third limb and active evaporation of
water after the valve. Finally, the silk thread exits the spider spigot with
post-spin drawing and further evaporation of water.
2
they encounter elongational flow, shear force and, simultaneously, changes in
their biochemical environment, including ion exchange, decrease in pH and
increased concentration.41,42 The pH reduction improves the shear sensitivity of
the dope to induce its gelation.43 Thus, simultaneous acidification together with
elongational flow are essential factors influencing the transformation from
dope solution to fibres.44 A simple protein denaturation model raised by Porter
and Vollrath proposes that the spin stress and pH gradient as factors to
unstable the amide–water hydrogen bond, and hence facilitate the replacement
of stronger amide–amide interaction.45 In addition, metallic cation changes
also contribute to promote the formation of silk fibre.46 For example, the K1
content is considered to induce the formation of b-sheet and nanofibril struc-
ture in spider silk gland dope.43,47 The liquid-crystalline spinning technology of
spiders significantly reduces the viscosity of the silk dope. Hence, it enables the
alignment of the protein molecules finely in a highly concentrated spinning
dope ,with high energy efficiency.3,42 The preliminary fibre is reached after an
internal drawdown process in the last limb of the duct, with active transport of
water after the valve.41,42 The biochemical and physical changes are accom-
panied by a liquid–liquid phase separation followed by a liquid–solid phase
transition that results in a preliminary silk fibre. Finally, the silk thread exits the
spider through the spigot with flexible lips, which places the fibre under tension
for the post-spin drawing, where the remaining solvent evaporates in air.48
Nature has created and refined protein structures through billions of years of
evolution for various specific purposes. Amino acid sequences and their folding
patterns make concerted efforts in creating elastic, rigid or tough materials,
making nature’s intricately designed products challenging examples for materials
scientists to develop bio-inspired materials. To mimic spider dragline silk suc-
cessfully, we need to duplicate the crucial features of feedstock dope as well as the
spinning process. The key issues include how the spider stores protein dope in a
highly concentrated liquid crystalline state and how it controls the processes in
the spinning duct to form a supremely tough thread. Molecular biology helps us
to extract, synthesize and assemble artificial genes to provide feedstocks for silk
production,27,49–51 whereas classical morphological studies discover the details of
the spider’s extrusion system.48,52–55 Combining these two biological disciplines
with process engineering can envision the design of advanced and benign fibre
extrusion technology, and eventually allow commercialization.
13.4 Mechanical Properties

One of spider silk’s advantages that attracts the increasing interest of scientists
is its outstanding mechanical properties. Natural silks typically have a larger
extension and higher toughness, although lower strength, compared to syn-
thetic fibres. Strength and extension respectively are the stress and strain at
which failure occurs. Toughness is calculated from the area under a stress–
strain curve, representing the sum of the energy absorbed by the material before
failure.9 As shown in Table 13.2, most synthetic fibres have restrained tough-
ness, although with high modulus and strength. In contrast, spider dragline
280 Chapter 13
Table 13.2 Tensile mechanical properties of spider silk and other well-known
natural and synthetic fibres.
Stiffness Strength Extensibility Toughness
Materials (GPa) (GPa) (%) (MJ m3) Ref.
Araneus diadematus MA 10 1.1 27 160 9
silk
Nephila edulis MA silka 7.9 1.15 39 165 58
B. mori cocoon silk 7 0.6 18 70 9
Nylon 6.6 5 0.95 18 80 9
Kevlar 49 130 3.6 2.7 50 9
Carbon fibre 300 4 1.3 25 9
High-tensile steel 200 1.5 0.8 6 9
Drawn at 20 mm s–1.25
a
silks with moderate modulus show toughness more than three times higher than
that of Kevlar 49 fibre, a type of synthetic fibre for bullet-proof vests. This
unrivalled toughness benefits from a combination of strength and extensibility.
Apart from its classical mechanical properties, dragline silk has the ability to
undergo supercontraction. This phenomenon will be elucidated in Section 13.6.
Furthermore, spider silk also has a torsional shape memory, which allows the
spider dragline thread, after being twisted, to oscillate only slightly, and by this
means to totally recover its initial form.56,57 This unique property allows spi-
ders to rapidly descend using dragline silk as a lifeline in case of danger.
13.4.1 Tensile Properties of Natural Silks

The mechanical properties of silks are determined from their response to tensile
deformations, which are conducted in machines such as the Instron Micro-
Tester,15,25 deriving stress–strain curves. As shown in Figure 13.3, typical
stress–strain curves show that both spider and silkworm silk fibres exhibit a
linear stress–strain relation until a yield point. The linear section of the curve is
the elastic region and the slope is defined as Young’s modulus.59 After the yield
point, the spider dragline initially exhibits a characteristic J-shaped behavior of
‘‘strain hardening’’ (the slope of the stress–strain curves increases with strain)
until point H, followed by so-called ‘‘strain weakening’’ behavior (the slope of
stress–strain curves decreases with strain) until the ultimate break. Other spe-
cies of spider draglines demonstrate similar stress–strain profiles,60 indicating
that strain hardening could be a general feature of spider draglines. On the
other hand, the silkworm silk acts only in the ‘‘strain weakening’’ mode in its
nonlinear portion of the stress–strain curve.
In addition to its capability of absorbing an enormous amount of impact
energy for a soft dragline filament, the impact force, on the other hand,
increases in a very slow manner owing to its marvelous extendibility and gra-
dually hardening mode. In contrast, Kevlar comprises very rigid molecules
Figure 13.3 A comparison of typical stress–strain curves of silk fibres from the spider
N. pilipes (by forced silking at 10 mm s1, curve a) dragline silk and
silkworm B. mori cocoon silk (curve b), Kevlar 49 (curve c) and rubber
(curve d). Inset: rescaled stress–strain curve of rubber. The stress–strain
curves of silk fibres (curve a and b) are divided into two parts: linear
(elastic) and non-linear regions. The mechanical characterizations were
performed at 22 1C and the humidity was kept at 50–55%. (Reproduced
with permission of Wiley-VCH from Du et al.25)
which incline to form mostly planar sheet-like structures61 whose filaments are
very rigid and brittle.62 If Kevlar 49 filaments become a substitute for the
draglines in a spider web, the collision of a flying prey into it would result in a
tremendous shock. As a consequence, the spider colonizing the web would have
been knocked off upon impact.
13.4.2 Factors Influencing the Mechanical Properties

of Natural Silks
The mechanical properties of silks can be utilized to probe the structures of
these materials. However, the mechanical testing results of silks are influenced
by several factors, such as forced reeling conditions, testing conditions, vapor
or solvent treatment, to different extents. Such factors are regarded as valuable
experimental and analytical tools to understand the structures of silks, instead
of simply as impediments to obtain reproducible results.
Most natural spider silks used in research were obtained by forced silking
technology, which is drawing the fibre from the spinneret of the immobilized
spider and simultaneously winding it on a computer-controlled spindle. This
282 Chapter 13
method is capable of controlling the structure and thus mechanical properties
of silks by tuning several key extrusion parameters, such as reeling speed, force,
temperature and reeling medium (e.g. into air or solvents such as water63).
Firstly, the spider dragline silks collected by forced silking at increased spinning
speeds would possess higher breaking stress and initial modulus, but lower
breaking strain.15,64 It is found that this can be attributed to the varied crys-
talline alignment in the silks reeled at different speeds, as observed by X-ray
diffraction.15,65 Besides, the silking speed has also been reported to influence the
size of the b-crystallites, which can also impact the mechanical perfor-
mance.15,66 In addition, the tensile performance of forcibly reeled silks can also
be correlated to the reeling force, because the silks exhibit distinct mechanical
properties when the reeling forces are different with all other parameters such as
reeling speed kept the same.63,67,68 Furthermore, the draw-down medium and
the length of the aqueous post-draw duct also play important roles in mod-
ifying the mechanical properties of a silk. For instance, spider dragline silk spun
into water displays greater strength, stiffness and resilience compared with silk
drawn into air, owing to the better alignment of the polypeptide backbones, as
discovered by Raman measurements. The resilience is defined as the ratio
between energy recovered and energy input in a loading–unloading cycle. The
difference in structure can be ascribed to the higher mobility and aligning time
that the silk proteins gain in an aqueous environment.63 Finally, the body
temperature would affect the mechanical properties of forcibly reeled spider
silks. An elevated silking temperature leads to an enhanced breaking strain and
almost constant breaking stress, hence an improvement in the silk toughness.58
This toughening effect may result from the temperature-induced changes in the
viscosity of the spinning dope, thus inducing a more relaxed conformation in
the amorphous regions when the spider silk forms.69
Testing conditions such as strain rate and measuring temperature also play
important roles in influencing the mechanical properties. Normally, when tes-
ted under higher strain rates, viscoelastic synthetic polymers such as nylon are
inclined to display increased stiffness and strength but decreased extensibility
and toughness. It is interesting to observe that the stiffness, strength, extensi-
bility and toughness of spider silk all increase with increasing strain rate.9 Even
measured at an extreme strain rate reaching 20–50 mm s1, Araneus diadematus
MA silks exhibit extensibility at 20–50% with the initial modulus at 25–40 GPa
and breaking stress at 2.0–4.0 GPa, thus obtaining toughness as high as
500–1000 MJ m3. According to the time–temperature equivalency principle,
increasing the strain rate and decreasing the temperature play similar roles in
affecting the mechanical properties of viscoelastic materials, such as silks. The
conditions of dynamic mechanical temperature analysis on MA dragline silk of
N. edulis spider at –60 1C enhance stiffness and strength, and breaking elon-
gation, compared with that measured at room temperature.70 Fractographic
studies show that spider silk behaves as a ductile material at temperatures as
low as liquid nitrogen temperature.70 When changing the measuring tempera-
ture in the opposite direction, increasing from room temperature to 150 1C, the
stiffness of the dragline silk decreases while the strength and extensibility almost
keep the same. Higher temperatures induce spider dragline silks to contract,
reaching up to 20% of their original length, indicating that the internal
hydrogen bonds are modified at higher temperatures.71 Further increasing the
temperature above 200 1C causes thermal denaturation of spider dragline silks
and hence reduces the intensity of the crystal diffraction and irreversibly
changes the lattice constants.71
Treating silks with polar solvents or vapor always greatly reduces the stiff-
ness and improves the breaking strain significantly.72–74 Polar solvents with
small molecules seem to outperform those with large molecules in influencing
the mechanical properties of silks. This is because the small molecules can
penetrate the silk more easily and also interact with polar groups of polypeptide
chains inside the silk more strongly, owing to their high polarity. Therefore,
water is regarded as a most active softener, followed by less active methanol,
while ethanol and butanol show almost no softening effect on spider dragline
silks which are immersed in them.75
Lee et al. revealed that metals can be successfully infiltrated into internal
protein structures of biomaterials such as spider silk through multiple-pulsed
vapor-phase infiltration (MPI) performed with equipment that is conventionally
used for atomic layer deposition (ALD). In their research, zinc, titanium or
aluminum, combined with water from corresponding ALD precursors, were
infiltrated intentionally into spider dragline silks, with the toughness of the
resulting silks greatly improved. The energy dispersive X-ray (EDX) and nuclear
magnetic resonance (NMR) spectra were used to confirm the presence of the
infiltrated metals such as titanium or aluminum inside the treated silks.76
In conclusion, factors such as speed, force, medium and temperature of the
forced reeling, strain rate, temperature of testing and vapor/polar solvent
treatment, can all influence mechanical properties of silks to different extents.
However, by tuning these factors, we can change the structure of the material,
leading to a difference in mechanical properties. In next section, we will discuss
the property–structure relationship to reflect a general view on how to control
the mechanical properties of silks by intrinsically tuning their structure.
13.5 Structure–Property Relationship

A deeper insight into the underlying structure–function correlation of silk
proteins is fundamentally important for the potential biomimetic production of
spider silk from recombinant and reconstituted silk protein with controlled
mechanical properties. Such knowledge also benefits novel synthetic polymer-
based materials, as spider silk organization can serve as a bio-inspired model
for designing sophisticated composite materials.
13.5.1 Structure Related to Mechanical Properties

Generally, the structural origin of the extraordinary extensibility of silk has not
been fully explained. Some ascribe the extensibility of the silk thread to the
284 Chapter 13
ability of the amorphous regions to stretch and move from their random coil/
other conformation into a more extended helix or b-strand configuration.9
Another model proposes the nanofibrillar morphology of the thread (i.e. in a
helical or a zigzag manner) as a major feature contributing to the extensibility
of silkworm silk.77 The morphology in a helical manner could contribute
because these helical fibrils would straighten parallel to the fibre as the original
silk threads extend and break. The morphology in a zigzag manner would play
a role because the zigzag fibrils would compress like a spring when the silk fibre
undergoes buckling or shear failure. Both effects could help to reduce failure
under a sharp bend while the former tensile effect could contribute to the well-
known large tensile strains of spider silks at breaking points.
Strength has normally been attributed to the presence of b-sheets packed into
crystalline regions and hydrophobic interactions within those crystalline
regions, which bind molecules together in the silk fibre.78,79 Crystalline regions
consisting of poly-Ala b-sheets are found to form more hydrophobic interac-
tions, and thus have a higher binding energy, than poly-Gly-Ala b-sheets.78
Given similar numbers and lengths of repeats, silks with poly-Gly-Ala repeats
with fewer interactions seem to have lower tensile strengths relative to those
with poly-Ala repeats. This is in accord with the lower tensile strength of MI
silk (with poly-Gly-Ala) compared with MA silk (with poly-Ala). It also agrees
with the lower tensile strength of silkworm silk (rich in poly-Gly-Ala blocks) in
contrast to spider dragline silk (rich in poly-Ala blocks). The higher crystal-
linity of silkworm silk compared to spider dragline silk may also account for its
brittle feature.
It appears that the overall toughness of silk threads is connected to its
structure at all hierarchical levels, including the secondary structure of poly-
peptide chain folding as well as the higher order structure of the protein con-
stituents. For instance, recent studies have proposed that spider silk filaments
can possess a complex ‘‘coat–skin–core’’ organization80.81 that is likely to
impart energy-dispersive properties.82 This structural complexity with micro-
fibrils83–85 enclosing fine channels80 is likely to contribute significantly to the
toughness of spider silk. Du et al. have illustrated how the intramolecular
b-sheet-induced strain-hardening contributes to the breaking energy, which will
be described in detail in the following section, covering many models raised to
address the issue of the structure–property relationship.25
13.5.2 Models
Many models have been proposed to interpret the mechanical properties of
natural silks from the perspective of structure. In Termonia’s model,12 the
dragline silk is described as many small stiff crystallites embedded in amor-
phous regions, which are made of rubber-like chains. The crystallites serve as
multifunctional interlock sites and create a thin layer of high modulus in the
amorphous region. The simulated properties of dragline silk are in accord with
the experimental results, while assuming the modulus of b-sheet crystals as for
fully extended crystals, i.e. 160 GPa. b-Spiral secondary structures formed by
the motif GPGXX in MA and flagelliform silk are suggested to act as springs,
being responsible for the extensibility of the silk.78 The proline residue would be
the focal point for the retraction energy after stretching. By forcing the proline
bonds to torque in response to extension, a large force can be generated for
retraction. It is proved by the evidence that the GPGXX motif only exists in
MA and flagelliform silks, which are also the stretchiest of spider silks. A
further support for the GPGXX motif providing the elasticity modulus is the
correspondence between the number of tandemly arrayed GPGXX repeats and
the different extensibilities of the two silks. MA silk, with up to 35% exten-
sion,86 has at most nine b-turns in a row before interruption by another motif.87
Flagelliform silk with 200% extensibility86 has at least 43 contiguously linked
b-turns in its spring-like spirals.88
In addition, mean field theory for polymers has been applied to predict
structure–property relationships in terms of chemical composition and mor-
phological structure, as well as the dynamic mechanical properties of silk,
derived in terms of the ordered fraction.89 This model predicts the range of silk
tensile properties in quantitative agreement with experimental results, and
suggests a numerical relation between the ordered fraction and in-water
shrinking capacity of spider silk. Zhou et al. has proposed a hierarchical chain
model to explain the nonlinear force–extension response of spider silks, and the
prediction well agrees with the experimental observations.90,91 In this model, a
polymer consists of many structural motifs, organized into structural modules
and supra-modules in a hierarchical manner, with their own characteristic force
for each module. The repetitive patterns in the amino acid sequence of the
flagelliform protein of spider capture silk give support to this model. A hairpin-
turn model is proposed based on studies of the molecular structure in silk,
which suggested that silk has a folded hairpin structure13,78,92 with about six
peptide segments per fold. In this model, three types of material in the silks are
defined as a ‘‘string of beads’’ and their approximate fractions are suggested for
silk nanofibrils, which determine the strength of natural silks.39,93 Typical
predicted stress–strain profiles at different reeling speeds for spider and silk-
worm silk are in good agreement with experimental measurements by imple-
menting the ‘‘b-sheet splitting’’ mechanism, and varying the secondary
structure of the protein macromolecules.94 The work by Buehler’s group95
highlighted the important role of hydrogen bonds contributing to the
mechanical property of silk. The work well explains why the nano-crystallites
can achieve high strength.
The molecular origin of strain hardening of spider dragline silk, compared
with rubber, Kevlar and silkworm silk, has been addressed by Du et al.25 The
occurrence of strain hardening originates from the unfolding of the intramo-
lecular b-sheets inside spider silk, which perform as ‘‘molecular spindles’’ to
store long molecular chains in space in a compact manner. With continuous
unfolding and alignment of proteins, the protein backbones and nodes of the
molecular network start to be stretched to support the load during fibre
stretching. As a result, the dragline filaments become progressively hardened,
286 Chapter 13
which enables efficient energy buffering as an abseiling spider tries to escape
from its predator through the safety line.
Specifically, during stretching, the intramolecular b-sheets inside the amor-
phous matrix will first unfold to release lengthy protein chains, with the b-crys-
tallites being unaffected. This will lead to a high extension of the dragline silk,
with the intermolecular linkages exempt from breakage. This process occurs as
the stretching reaches the yield point S in Figure 13.3. At the beginning stage of
protein unfolding, the modulus declines to nearly zero as the breaking of weak
intramolecular hydrogen bonds makes the major contribution to fibre elonga-
tion. As the elongation continues, the progressive unfolding and alignment of
protein chains result in stretching of the protein backbones and nodes of the
molecular network to support the load. As a consequence, the dragline silk thread
becomes stiffer owing to the contribution coming from enthalpic component.96
These processes exhibit how strain hardening happens in spider dragline silk.
Similar unfolding behavior in recombinant spider silk protein97 and some other
elastic proteins98 are also shown by single protein molecule stretching of these
proteins using atomic force microscopy (AFM). The further stretching exceeding
the inflection point H will generate breaking of the b-crystallites, destroying the
interlocks of the molecular networks with the silk threads, thus giving rise to the
weakening of the silk filaments (also termed as strain weakening). The process of
b-crystallite breakage in the spider dragline silk is verified by the XRD mea-
surements, demonstrating that once a spider dragline silk is stretched beyond
point H, the total b-crystallinity of the dragline will decrease correspondingly
(Figure 13.4). In contrast, there are much fewer intramolecular b-sheets in the
amorphous region of B. mori silkworm silk. Therefore, silkworm silk only exhi-
bits strain weakening after the yield point, with inferior extensibility.
The response of spider dragline silk upon stretching is found to deviate
greatly from what occurs in rubbers.99 As revealed in Figure 13.4, instead of
inducing crystallization like that in rubber, the tensile deformation of spider
draglines results in breaking of the crystallites, giving rise to the decrease of
crystallinity shown by the XRD measurements. The distinction between the
behavior of silk and rubber can be credited to their disparity in molecular
structure and intermolecular connections established upon the formation of the
material. The nano-sized b-crystallites together with the orientated amorphous
region3,100 in spider silk are formed during the silk-spinning process in the
animal’s sophisticated spinning organ known as the spinneret.3,101 During the
silk-spinning process, substantial shearing is applied to the silk protein mole-
cules to promote the alignment of the adjacent protein molecules.3,101–103 The
b-structures (polyA or polyGA regions) can thus be crystallized into nano-b-
crystallites.3,15 Once the silk fibre is formed, the protein chains inside are
unlikely to form new b-crystallites by stretching. On the contrary, rubbers,
consisting of entirely random-oriented molecular chains possessing the same
subunit,104 can smoothly form crystallites while the amorphous chains are
aligned under stretching.99
The different structural characteristics in spider dragline silk and rubber also
result in their remarkably different recovery dynamics from a stretch. Rubber
Figure 13.4 Effect of tensile deformations on the b-crystallinity of spider N. pilipes

dragline fibres. The lower curve is the first derivative line of the stress–
strain curve of the spider draglines (upper line, 12 samples averaged).
(Reproduced with permission of Wiley-VCH from Du et al.25)
will oscillate severely, with the amplitude declining to zero at a slow speed after
being stretched.25 Spider dragline fibre, on the other hand, decays rapidly to
equilibrium, with small oscillations. This demonstrates that spider dragline silk
rarely oscillates after a disturbance like elastomers such as rubbers, with similar
behavior also observed in a torsional stability study of spider dragline silk.56
The different recovery responses between spider dragline silk and rubber
indicate an optimized viscoelasticity in silk protein molecules, which can be
attributed to the special organization of molecular chains in spider dragline
silks. The features encompass a more compact packing of molecules by folding
into intramolecular b-sheets or crystallizing into b-crystallites, and a higher
degree of molecular orientation in comparison to rubber. As a result, spider silk
is capable of absorbing energy without generating a drastic oscillation like the
way elastomers behave.
The dragline silk fibres extruded at faster reeling speeds have larger strain-
hardening-induced strain, because they contain an increased content of the
intramolecular b-sheets compared with those drawn at lower speeds.25 The
strain-hardening-induced strain is defined as the strain produced in the strain-
hardening curve segment (the strain difference between points S and H). The
strain-hardening strain ratio is defined as the strain-hardening induced strain
over the total strain of the non-linear region. The strain-hardening strain ratio
is found to grow substantially with the ratio of intramolecular b-sheets to the
total content of b-sheets.25 As shown in the experimental results, the crystal-
linity decreases from 0.22 to 0.11 when the reeling speed increases from
10 mm s–1 to 200 mm s–1, as is indicated from the XRD measurements. The
content of the intramolecular b-sheets rises from 0.29 to 0.45, while the overall
288 Chapter 13
content of the b-sheets keeps almost the same (from 0.51 to 0.56), as shown by
FTIR spectroscopy.25
The underlying mechanism could be explained by the well-controlled
formation process of spider dragline silk.25 An escaped nematic liquid
crystalline phase exists in the silk gland and in the first half of the duct of the
spider, in a tight hexa-columnar packed arrangement of compactly coiled
molecules.23,105 The flow elongation and the exchange of ions along the duct
are thought to encourage the molecules to partly unfold. The retention of
some hairpin loops results from the incomplete extension of the molecules. It
would help to provide an anti-parallel arrangement,96 which could facilitate
the formation of b-structures with extension flow. More of such hairpin
loops are retained in the duct when the silk fibres are extruded from a spider at
high speeds, as they do not have sufficient time to respond to the fast external
drawing. As a consequence, the content of the intramolecular b-sheets rises,
and thus the strain-hardening effect becomes more significant with the faster
drawing of the fibre, while the overall content of the b-sheets keeps almost
the same.25
13.5.3 Techniques to Study the Structure of Silk Fibres

In this section we summarize and compare several techniques that are widely
used to characterize silk fibres, revealing the general morphological and
structural characteristics in their static or dynamic process.
Firstly, the microscopic morphology can be observed by scanning electron
microscopy (SEM). AFM can be utilized to scrutinize the topography of fibres
such as B. mori silkworm silk and spider silk,106 offering details on the nan-
ometer scale. Analytical transmission electron microscopy (TEM) can
demonstrate the diffraction patterns from organized structures in a fibre and
assess the crystallite content in dragline silk.107
Secondly, advanced imaging techniques such as Raman spectroscopy, FTIR
spectroscopy, XRD and NMR are important tools for analyzing the secondary
and tertiary structure of silkworm and spider silk threads.
Raman and FTIR spectroscopy are non-invasive techniques used to inves-
tigate the dominant conformational content of the fibre, by monitoring the
amide I, II and III frequencies values, which are indicative of a-helix, b-sheet,
random coil and turns in the fibre.75,108,109 The advantages of Raman spec-
troscopy over FTIR involves the low interference by water in the natural bio-
fibres and the chance to acquire spectra of single fibres with a diameter of less
than 10 mm.110 Raman microspectroscopy was utilized to obtain the orientation
of the carbonyl groups in silks via the amide I band by recording polarized
spectra, therefore determining the orientation of the b-sheet crystallites.110
Conformational and orientation changes in fibres under stress and strain can
also be observed by Raman spectroscopy.111–113 Raman spectroscopy on single
silk threads during gradual stretching shows that the silk structure under-
goes uniform stress during deformation,72,111,114 suggesting the presence of
micro-fibrillar structures within the silk thread that evenly distribute any
applied stress.
XRD can also be applied to characterize the structure of silk threads during
gradual stretching together with the structure in the static state. Wide-angle
X-ray diffraction (WAXD) and small-angle X-ray scattering (SAXS), when
combined with synchrotron radiation, can be used to characterize small sam-
ples such as single fibres. The interference patterns coming from diffraction of
the radiation can be related to the distance between domains in the fibre.
Therefore the patterns can reflect the relative arrangements of the domains.
A length scale similar to the radiation wavelength (B1 Å) is probed by this
technique, discovering information of fibre structure such as crystallinity,
crystal size, inter-crystalline distance and orientation distribution. Varying
alignments of crystalline structure, crystallinity and crystal size in the silks
reeled at different speeds were observed by XRD studies.15,115–118 XRD pat-
terns obtained from single threads being spun119 also revealed that the thread
consists of small crystalline blocks in a matrix containing both oriented and
disoriented amorphous material, with the crystalline fraction having a
b-poly(L-alanine) structure.117
Solid-state NMR (SS-NMR) can characterize the dynamics and reveal the
local conformation where specifically labeled amino acids are discovered. The
structural information gained by SS-NMR depends on the measurement of
chemical shifts, torsion angles and inter-nuclear distances. The packing
arrangement and size are acquired through experiments which probe the
dynamics and manifest information on the crystalline and non-crystalline
content. For example, proton-driven 13C two-dimensional NMR spin-diffusion
experiments on dragline silk thread suggested that the poly-Ala segments adopt
a highly ordered b-sheet structure13,120 and the glycine-rich segments have 3(1)
helical structures,14,121 both oriented in line with the thread. However, it
demands a greater amount of sample and isotopic labeling of the fibre in most
situations. For a biological fibre, SS-NMR does not allow sequencing a specific
amino acid assignment by itself, nor does it provide a complete structure.
In comparison, microscopy-related techniques are used to image fibre mor-
phology, while Raman and FTIR spectroscopy can give a global portrait
conformational content in the fibre. The XRD techniques display information
in the crystalline regions of a material.
SS-NMR, as a powerful technique to study natural protein fibres, allows the
study of the molecular structure as well as the dynamics of semi-crystalline and
amorphous material.120,122–130
13.6 Supercontraction
Supercontraction, as its name suggests, is a phenomenon describing the large
shrinkage in the length of unrestrained spider dragline silk fibre when it is
immersed in water or exposed to high relative humidity environments at room
temperature.131–133 It is a distinctive feature of the MA and flagelliform silk
290 Chapter 13
3,134
fibres spun by orb web-building spiders. When wetted in an axially
unrestrained conditions, spider dragline silk retracts up to 50% of its original
length, while increasing in overall volume, decreasing in stiffness by several
orders of magnitude and causing a dramatic change in its mechanical beha-
vior.131,132 This process generates substantial stress in restrained fibres, owing
to restricted supercontraction, in excess of 50 MPa131,133,135 and amounts up to
4.7% of the breaking strength.131,136 Wetting-induced supercontraction of
spider dragline silk is presumed to be exploited to take up slack in webs and
restores web shape and tension after deformation by prey capture, precipitation
or wind as they become loaded with dew or rain.131–133,135,137–139 This shape
memory behavior of dragline spider silk is unique in that it is brought about
totally in a benign condition at room temperature when wetted with moisture.
Similar shape memory behavior can also be observed for some polymers, but
only induced by external stimuli such as high temperature or high pressure or
harsh solvent conditions.100,140,141
Dry fibres, either forcibly silked or naturally spun fibres, behave as glassy
polymers: there is an initial large modulus followed by a linear-elastic regime up
to a yielding point, with a subsequent decrease in the slope of the stress–strain
curve to a minimum and, lastly, accompanied by a gradual increase in the slope
until breaking point.133 In contrast, the tensile behavior of supercontracted silk
tested in water seems to correspond to an elastomeric material: an initial low
modulus and large ultimate strain. This difference in the mechanical properties
between dry and supercontracted spider silk fibres has been modeled through a
double network of hydrogen bonds and protein chains.142 The model assumes
that supercontracted fibres correspond to a state where the hydrogen bond
network is disrupted and the observed elastomeric behavior tensile properties
are controlled by the alignment of the protein chains. The elastomeric behavior
only appears in wet supercontracted fibres since drying freezes the conforma-
tion of the chains by re-establishing the hydrogen bond network.12,142,143 These
conformational changes are reversible when the humidity is increased and when
chain reorientation is allowed.139
Research carried out by Gosline10 revealed that supercontraction has pro-
found implications on the mechanical behavior of spider silk, even hypothe-
sizing a relationship between supercontraction and tensile properties.12,142 It is
found that the whole range of variability of the tensile behavior of spider silk,
either naturally spun or forcibly silked, can be predictably and reproducibly
tailored through a combination of supercontraction and stretching, suggesting
that the variability corresponds to different degrees of alignment of the protein
chains.133 Controlled supercontraction may thus be exploited as a mechanism
to tailor the properties of spider silk, allowing it to be implemented in the
operation of spinning artificial silk fibres in large quantities for different bio-
mimetic applications, as well as improving the reproducibility of the fibre
tensile properties.133,144
Some authors found that supercontraction stress was transient and that silk
fibres lose tension within 200 s when the humidity remains high, suggesting that
supercontraction is ephemeral and supercontraction stress could not maintain
135
tension in wet webs. They argued that this stress relaxation is a major
impediment to technological applications of spider silk that are based on the
protein and network structure of spider dragline silk. Subsequently, Savage
et al.145 replied that the stress relaxation was largely an artifact of Bell et al.
using forcibly silked material, rather than naturally spun silk, since spiders
impart a large frictional force on forcibly drawn silk through their internal
friction brake.67 In contrast, others demonstrated no signs of stress relaxation
in forcibly silked dragline silk, despite it belonging to both different and similar
species.133,145,146 They showed that supercontraction tension can be sustained
for long periods of time, suggesting a permanent change in the molecular
structure within the silk fibre. Therefore the hypothesis that supercontraction
maintains tension in the web after deformation by water restoring its shape
is clearly supported in these studies. However, the supercontraction stress
generated varied widely between studies by an order of magnitude
(10–140 MPa133,136,145–147 and 300–400 MPa135), perhaps due to the different
species sampled,64,136,148–150 possibly related to the differences in content of the
critical amino acids such as proline,22,59 spinning effects resulting from varia-
tion in forces applied to the fibre by the spider during artificial silking,67
spinning conditions on fibre orientation100 and external factors. Agnarsson
et al. demonstrated that the observed phenomenon in the variability of
supercontraction force depends upon the rate of increase in relative humidity147
to the threshold level of humidity causing supercontraction. Rapid increases in
humidity (41% s–1), over a few seconds, generated supercontraction stresses
exceeding 100 MPa, whereas slower increases in humidity, over several minutes,
led to relatively low supercontraction stresses not exceeding 80 MPa in super-
contracted forcibly silked dragline fibres.
It is known that spider dragline silk can be modeled as a semicrystalline
material, consisting of multiple fibroins joined by polyalanine nano-b-crystal-
lites embedded in a non-crystalline amorphous network.10,15,16,78,86 Spider
dragline silk is composed mainly of two proteins, spidroin I and II, which can
be divided into block copolymers with GAGA(A)n blocks and glycine-rich
domains.18 The amorphous network consists of relatively ordered glycine-rich
linker domains and proline-containing random coils. The glycine-rich linker
domains that retain fibroin orientation before supercontraction mostly occur in
the major ampullate spidroin I (MaSp I), whereas the random coils are pro-
duced by the greater abundance of proline in MaSp II.151 These two proteins
are partially phase-separated during fibre formation, such that they are dis-
persed heterogeneously throughout the silk, with MaSp II occurring largely in
the interiors of threads and MaSp I dispersed throughout.152 Infiltration of
water during supercontraction is hypothesized to plasticize the silk fibres by
disrupting hydrogen bonding within the amorphous region of the proteins,
thereby increasing molecular mobility, allowing a subsequent entropy-driven
reorientation and coiling of silk molecules into a less organized configura-
tion.10,73,121,133,145,153,154 Water molecules exert a similar plasticizing effect on
silkworm silk.143 Recent studies have focused on the importance of disrupting
secondary structure in the glycine-rich blocks for mobilization of the proteins
292 Chapter 13
22,92,149
within the amorphous network. Hydrogen bonding within the random
coils is relatively weak and disrupted by small amounts of water, while the
stronger hydrogen bonds of the glycine-rich linker regions are partially dis-
rupted at high humidity.133,145,146 This disruption in secondary structure is
sufficient to reconfigure the silk protein network, driving the reorientation of
the silk molecules and allowing the silk proteins to move rapidly to more dis-
ordered and higher entropy configurations. This causes a sudden contraction in
the entire unrestrained silk fibre while expanding in overall volume, behaving
like a filled rubber with a relatively low modulus, or generates substantial
stresses in the restrained silk.146
Dragline silk exhibits a cyclic contraction phenomenon distinct from
supercontraction: it extends and contracts repeatedly like a muscle, by cycli-
cally raising and lowering the relative humidity (Figure 13.5).146 Cyclic con-
traction is repeatable both before and after supercontraction. This cyclic
response produces high forces, exceeding supercontraction forces, which are
completely reversible and that can be precisely controlled through humidity
alone. Agnarsson suggested that spider silk emerges as an attractive model for
biomimetic muscle fibres as it generates a power density 50 times higher than
biological muscle.155 Water binds to silk permanently during supercontraction,
resulting in a change in its structure, whereas cyclic contraction results from a
reversible loss of water during drying. This cyclic phenomenon is not explained
by molecular models developed to explain supercontraction.10,22,154,156 Initial
exposure to humidity results in infiltration of water molecules into the silk fibre.
The water molecule first binds itself to the random coil within the amorphous
Figure 13.5 Cyclic contraction of spider dragline silk. (Reproduced with permission
of the Company of Biologists from Blackledge et al.146)
region, disrupting the relatively weak hydrogen bonding. This relaxes the silk
and explains the initial dip in the stress–strain curve when silk is first expose to
humidity just before the critical threshold of 70–75%. When the humidity
reaches the critical threshold level, most of the hydrogen bonds within the
random coil have been disrupted. Now the water molecules start to bind to
the oriented glycine-rich linkers within the amorphous region, disrupting the
relatively strong hydrogen bonding. This disruption in hydrogen bonding
within oriented glycine-rich linker regions increases the local mobility of
molecules, allowing them to reconfigure to a less organized state that shrinks
the fibre and traps the water molecules in the silk structure, causing super-
contraction.59,121 Upon drying immediately after supercontraction, the water
molecules are lost only from the random coil region, rebuilding hydrogen
bonds in the process that immobilize the silk molecules, contracting the fibre.
Subsequent wetting results in water molecules binding to the random coil
region, breaking the hydrogen bonds and causing the silk fibre to relax. In
short, high humidity in cyclic contraction causes the silk to relax if the mobi-
lization of silk molecules enabled by moisture is limited to the random coils
region within the silk, by breaking the relatively weak hydrogen bonds.
However, high humidity in supercontraction causes the silk to supercontract
and hence manifest itself as an increase in tension in restrained silk fibre, if the
local mobility of silk molecules enabled by moisture is limited to the oriented
glycine-rich region within the silk, by breaking the relatively strong hydrogen
bonds.146 Thus supercontraction is a two-step process that depends on the
wetting-induced mobilization in two different regions within the amorphous
region: oriented glycine-rich linkers and random coil. Cyclic contraction is also
observed in silkworm silk, as well as natural fibres such as cotton and hair,
whereas it is not observed in hydrophobic synthetic fibres such as polyethylene
or acrylic.155 This suggests that cyclic contraction depends on the hydrophilic
nature of the constituent amino acids. The contraction stress generated is
16 MPa in silkworm silk,157 0.6 MPa in cotton, 6.1 MPa in hair and an
estimated 60 MPa in dragline silk.155
Supercontraction and cyclic contraction generate substantial forces, pro-
viding a possibility that these phenomena could be exploited to perform work.
Contrary to popular belief that supercontraction is a one-time response and
hence a limit in its application to perform work, Blacklege et al.146 have
demonstrated that supercontraction has a multiple occurrence within the same
dragline silk if the fibre is re-extended to its original length before drying after
each unrestrained supercontraction. This happens because physical energy
provides energy to the silk, reconfiguring the silk proteins within the amor-
phous region to a more ordered state. Once dried, the reinstating of the
hydrogen bonds maintains the structural organization of the silk proteins.
Subsequent wetting increases the mobility of silk proteins in different regions,
reconfiguring the silk proteins to a higher entropy state, driving the super-
contraction of the fibre.146
In summary, spider silk makes use of supercontraction to maintain tension in
the web and restore web shape after deformation by prey capture, precipitation
294 Chapter 13
or wind as they become wetted with morning dew or rain. The powerful cyclic
contraction exhibited by spider silk also results in a green and energy efficient
mimicry of biological muscles. The multiple occurrences of supercontraction
and fatigueless cyclic contraction offer possibilities for performing work in
industry and in clinical sciences.
13.7 Silk Protein-based Novel Biomaterials

One appealing application of spider silk is to mimic the disparate material
functions of the proteins to design novel biomaterials. Efforts have been
continually focused on studying the assembly and processing of spider silk
proteins into diverse material forms to expand applications to medical, elec-
tronic and optical fields. Owing to its biocompatiblity and controllable
degradability, the silk protein material can be promisingly anticipated as use in
sutures, drug delivery systems, fibre-based tissue, flexible electronics for phy-
siological recording and functions, and optical systems for diagnosis and
treatment.158–161
For example, scientists are investigating spider silk as ‘‘artificial skin’’ to
replace the skin from a body for a graft. The spider silks are biocompatible and
controllably biodegradable, meeting the requirements of a graft. Compared
with other materials, they are also strong and tough enough to resist all the
rigors that ordinary skin would experience. Wendt et al. successfully cultivated
skin cells on spider silk meshes to replace lost skin tissues, forming equivalents
of the epidermis, the outermost layer of the skin, and the dermis, the living
tissue below the epidermis containing blood capillaries, nerve endings and other
structures.158
Apart from pure silk proteins for materials, protein block copolymers and
chimeric proteins will become more available. The most extensively studied
recombinant spider silks encompass those from N. clavipes (MaSp I and
MaSp II), A. diadematus (dragline ADF-3 and ADF-4) and N. clavipes
flagelliform.26–28,30,32,37,162–164 Much effort has been exerted to explore a range
of different host systems for large-scale expression of gene coding spider silk,
containing yeast, insect cells, bovine mammary epithelial alveolar cells, baby
hamster kidney cells, transgenic plants such as Arabidopsis, soybean, potato
and tobacco, transgenic mammals such as mouse and goat, and transgenic
silkworms.26,28,32,37,162–169
Native spider silks and recombinant silk proteins can be solubilized with
solvents, including formic acid, hexafluoroisopropanol (HFIP), calcium nitrate,
lithium salts or ionic liquids, then processed into different structures covering
fibres, films, gels, porous sponges and other related systems.170–172 Techniques
utilized to form fibres involve solvent extrusion and microfluidic and electro-
spinning methods. Solvent extrusion refers to drawing the thread through a
coagulation bath in a controlled way. Microfluidic methods modulate the
geometry and chemistry of the outcome fibre by contracting channels and
diverse solvent inputs. Electrospinning processes integrate strong voltage
gradients with syringe pump extrusion to fabricate either random or aligned
fibre deposition.173
Spider silk fibres of 10–60 mm diameter have been fabricated through the
electrospinning of genetically engineered silk spin dopes in urea containing Tris
buffer and salts. This aqueous method has generated fibres similar to natural
spider silk in terms of molecular orientation and mechanical properties.174
Smaller fibre diameters up to 40 mm can be formed through electrospinning of
spin dopes derived from ADF-3 recombinant spider-silk protein. This method
generates fibres with similar toughness and modulus, but with lower tenacity
compared to native A. diadematus dragline silk.37 Fibre diameters down to
300 nm can be generated through electrospinning of N. clavipes MaSp I silk
dope prepared in HFIP. Thus fibre properties can be potentially improved
through the control of conformation and molecular alignment.175
Water-insoluble, transparent films with thickness in the range 0.5–1.5 mm can
be obtained by casting solutions that contain recombinant forms of the dragline
spider silk proteins ADF-3 and ADF-4 through the use of potassium phosphate
or methanol. This treatment promotes the conversion of the protein’s sec-
ondary structure from a-helix to b-sheet, creating a level of chemical stability in
certain denaturants that is comparable to that of native dragline silk. Various
cloning techniques and the approach of modifying film surfaces with functional
molecules envision these materials as promising biomedical and clinical
applications, such as wound dressings and scaffolds.176,177
Hydrogels with excellent superior mechanical response and stability over
weeks are utilized in the field of tissue engineering to form porous but stable
tissue scaffolds.178 Silk nanofibres of diameter B3 nm and lengths less than 1
mm appear after the addition of methanol to recombinant forms of the dragline
silk protein ADF-4, and then transform into a hydrogel fibre network over
several days. These hydrogels demonstrate a non-linear viscoelastic material
response with low stiffness and strength, which can be improved by a cross-
linking process that will promote a hydrogel with a fairly linear material
response, a much greater modulus and strength response. Cross-linking can be
induced by visible light and treatment with ammonium peroxodisulfate and
tris(2,2 0 -bipyridyl)dichlororuthenium(II).
Microcapsules with high mechanical stability and controlled properties of a
constrained degradation response to tissue-specific enzymes can be developed
by controlling the silk self-assembly at an oil–liquid interface of the recombi-
nant form of ADF-4 spider silks. Therefore, these materials show promising
applications, ranging from drug delivery to microreactor design.179
Surface functionalization has been applied to silk surface chemistry to
influence cell and tissue development through targeting carboxylic acid groups
on the amino acids in the protein.170 Spider silk has a significantly higher
number of carboxyl side groups on the amino acids compared to silkworm silk,
which provides higher degrees of cell-modifying functional groups.180
EDC [1-ethyl-3-(3-dimethylaminopropyl)carbodiimide] has been used to
functionalize ADF-3 and ADF-4 garden spider A. diadematus dragline silks by
coupling films formed from the proteins with fluorescein and b-galactosidase
296 Chapter 13
176
using carbodiimide chemistry. This approach could be used for biosensors
and reactive media and to influence cell and tissue functions by increasing high
levels of surface decoration with enzymes.
Additional benefits of providing designed functionalities encoded during
gene construction can be provided from genetically engineered spider silks.
Incorporation of cell binding domains, such as RGD (Arg-Gly-Asp),109 or
domains that interact selectively with inorganic components, are some strate-
gies employed for N. clavipes spider silks to generate novel organic–inorganic
composite material systems.181,182 Such incorporation allows the highly tai-
lored hybrid or chimeric spider silks to selectively undergo mineralization with
silica181 and hydroxyapatite.12 By controlling the processing conditions, the
morphology and structure of the composite could be manipulated. A biomi-
metic approach toward tougher and stiffer silk composite materials can be
achieved by controlling the size of the inorganic domains, as well as the
molecular-level interactions between the organic and inorganic domains in
these composites. These results suggest that chimeric spider-silk proteins might
provide new uses for spider silk in biomedical and other specialty materials,
such as in hard tissue remodeling or in adhesive fillers. The ability to process
and assemble these novel composite materials in aqueous ambient processing
conditions is another benefit of this approach.
13.8 Summary
In summary, we have reviewed the inherent properties of spider dragline silk as
well as the technological advances on the possible applications of dragline silk
in the biomedical and clinical fields.
Spider dragline silk exhibits excellent mechanical properties in terms of
breaking energy, coupled with both high strength and strain at failure. The
mechanical properties could be further tuned by varying external factors such
as strain rate, temperature, reeling speed and force, the medium of the forcibly
draw silk, and treatment with solvents and vapors. In addition, spider silk
utilizes its inherent property, supercontraction, to maintain tension in the web
and restore web shape after deformation by prey capture, precipitation or wind
as they become wetted with morning dew or rain. The powerful cyclic con-
traction exhibited by spider silk also results in a green and energy efficient
mimicry of biological muscles. The multiple occurrences of supercontraction
and fatigueless cyclic contraction offer possibilities for performing work in
industries and clinical sciences.
The outstanding mechanical properties of spider silk are determined by its
unique structural characteristics formed through its incomparable dope storage
and spinning process. Scientists have applied many techniques in order to study
the structure of the dragline silk. SEM and AFM are employed to study the
morphology and topography of the fibre, and WAXD and FTIR are used to
assess the b-crystallite and total b-sheet content in dragline silk, respectively.
Besides, NMR, Raman spectroscopy and SAXS are also important tools in
analyzing the secondary and tertiary structures in dragline silk. Generally
speaking, dragline silk can be modeled as a semicrystalline material, in which
the polyalanine nano-b-crystallites are embedded in the non-crystalline amor-
phous region to form a network. In particular, the model by Du et al.25 sug-
gested that intramolecular b-sheets acting as ‘‘molecular spindles’’ addressed
the molecular origin of strain hardening of spider silk filaments.
Last but not least, reconstituted and recombinant silk proteins can be used to
fabricate novel biomaterials for application in the biomedical and clinical fields.
The proteins are processed into different structures ranging from fibres, films,
gels, hydrogels and porous sponges to microcapsules. Surface functionalization
through targeting carboxylic acid groups on the amino acids in the protein
could be used for biosensors and influence cell and tissue functions by
increasing high levels of surface decoration with enzymes.
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CHAPTER 14
Mussel Byssus Fibres: A Tough

Biopolymer
F. G. TORRES,* O. P. TRONCOSO AND C. E. TORRES
Department of Mechanical Engineering, Catholic University of Peru,

Av. Universitaria 1801, Lima 32, Peru
14.1 Introduction
Byssus fibres produced by mussels are tough biopolymers composed mainly of
proteins and water. These natural biopolymer fibres have been intensively
studied owing to their mechanical and adhesive properties. The remarkable
strength, unmatched toughness and extensibility of byssus fibres allow mussels
to withstand the large and repetitive forces produced by crashing waves with
velocities of over 10 m s1 and accelerations of approximately 400 m s2. They
are composed of three collagenous proteins that make up the bulk of the thread
core. Their toughness is considered six times greater than that of the human
tendon collagen and comparable with that of Kevlar and carbon fibres.
In addition, the adhesive properties of byssus fibres allow mussels to be
attached to hard and soft substrates in aquatic environments through their glue
proteins located in their adhesion plaques. These plaques contain at least six
different types of adhesive proteins that have attracted great interest for their
potential applications.
In this chapter, we review the properties, characterization and applications of
byssus fibres. First, we give a brief introduction to the natural history of
mussels. Then, we discuss the structure at different levels of byssus fibres,

305
306 Chapter 14
including their chemical composition and morphology. We then continue by
describing the structural, mechanical, thermal and adhesive properties of
byssus fibres. Finally, we present several applications of the mussel adhesive
proteins found in the byssus fibres. The last section includes the analysis of
adhesive properties for applications in medicine and biotechnology, such as cell
culture, tissue engineering, medical, dental and biosensor applications. All
these aspects are discussed in order to present the potential use of byssus fibres
that suggest new models for the design of novel biomimetic polymers.
14.2 Natural History

14.2.1 Biological Functions
Mussels live in a variety of habitats, such as rocky intertidal, salt marshes and
subtidal and hydrothermal vents. Byssal fibres provide mussels with a secure
attachment to the rocks present in their aquatic habitats.1 These fibres present a
combination of strength and extensibility, together with a low resilience, that
give them an unmatched toughness that allows mussels to survive the large and
repetitive forces produced by waves.2 With gentle waves, the byssus fibres
provide an elastic response, which allows mussels to move freely with the
waves. With violent waves, byssal threads are able to dissipate energy and
increase their attachment strength.3 Without these properties, mussels would be
dashed against rocks by the waves.
14.2.2 Mussel Anatomy

Mussels are aquatic invertebrate animals living in freshwater and marine
habitats. They belong to the phylum ‘‘mollusca’’ under the class ‘‘bivalve’’.
Their anatomy is characterized as having two shells or valves that are joined
together by a ligament and are closed by strong internal muscles. Internally,
mussels have a large organ called a foot that is used for mussel locomotion.
Byssus fibres are secreted by this ‘‘foot’’ and are responsible for tethering the
mussel to the rock surface via some adhesive plaques (Figures 14.1a and 14.1b).
Three main glands are located in the mussel foot: the stem forming gland, the
thread forming gland and the plaque forming gland. The first gland forms the
stem, an organic tissue that supports each byssus fibre serving as a connection
between the foot and the byssus fibres (Figure 14.1). Byssus fibres are generated
from the stem located in the mussel foot in an injection moulding-like process.4
As shown in Figure 14.2, the byssal fibres are located all along the stem. The
fibres attach to the stem with a hoop of thin tissue. An adhesive plaque is
located at the end of each fibre. The total number of fibres along the stem varies
from mussel to mussel, and increases with water turbulence.5 Large numbers of
threads allow mussels to attach to several surfaces.
Mussel Byssus Fibres: A Tough Biopolymer 307
(a)
(b)
Figure 14.1 (a) Schematic view of a mussel showing the stem, byssal thread and
adhesive plaque;8 (b) photograph of byssus fibres from Aulacomya ater
attached to various stones.
14.2.3 Mussel Species

Byssal threads of a broad range of mussel species from diverse environments
have been studied. Intertidal and subtidal mussels in freshwater and marine
habitats such as Modiolus modiolus, Geukensia demissa, Bathymodiolus ther-
mophilus, Dreissena polymorpha, Aulacomya ater and Perna canaliculus are
among these species.1,6–8 Byssus from the genus Mytilus, such as Mytilus edulis,
308 Chapter 14
Figure 14.2 SEM micrograph of a stem and its byssus fibres from the species
Aulacomya ater.
M. californianus, M. galloprovinicialis, M. coruscus and M. trossulus, has also

received considerable attention.1,9–14 The results show that byssal fibres from
this genus have a similar composition and structure.
However, the threads of byssus produced from different species of mussels
are not equivalent.1 The differences found in the mechanical properties of
mussel byssus from different species are probably related to adaptations that
help mussels survive in different environments.15 Flow, salinity and tempera-
ture are among the environmental factors studied for their influence on the
mechanical properties of mussel byssus.16–18 As an example, the byssus fibres of
M. californianus are stronger than those of M. edulis.19,20 The adhesive
attachment of M. galloprovincialis is stronger than that of M. edulis.21,22 Even
the season and wave exposure are factors that affect the attachment strength
within specific species.20,23,24
14.3 Morphology and Structure

14.3.1 Chemical Composition
Table 14.1 shows the identified constituents of mussel byssus. They are com-
posed mainly of proteins and water. In the case of M. edulis, freshly collected
byssal threads are about 70% water, by weight. Of the dry weight, protein is
about 96%.25 Small amounts of inorganic residues such as ash, heavy metals
and transition metals have also been found in mussel byssus.26
Table 14.1 Compositional analysis of M. edulis byssal threads.
Percentage (%) Ref.
Water content (% total wt) 70 3 46
Dry weight (% total wt) 30 2 46
Protein (% dry wt) 95 3 25
Ash (% dry wt) 4.8 46
Cu/Fe/Zn (% dry wt) 0.1 26
Other cations (% dry wt) 0.2 26
The byssus fibre is primary composed of collagenous proteins known as ‘‘pre-

pepsinized collagens’’ (PreCol) that make up the bulk of the thread core.25,27–29
The adhesive plaque located at the end of the fibre is composed of at least six
different types of proteins known as ‘‘foot proteins’’ (fp). These proteins are
labelled according to their specific mussel species. As an example, five dis-
tinctive types of foot proteins have been isolated from the adhesion plaque of
M. edulis and are labelled from Mefp-1 to Mefp-5.30–34
14.3.2 Macro Structure of Byssal Threads

The formation of byssal threads begins when the mussel foot touches down
on a surface that the mussel finds suitable for attachment.4 Soluble thread
precursors are secreted into the ventral groove of the foot, where muscular
contractions mould them into functional threads. This process lasts around
5 min.15 The grooves depicted in Figure 14.3 are a consequence of the injec-
tion moulding-like process that forms byssal threads as they are secreted in the
liquid state and moulded against the walls of ventral grooves.35
The byssus fibres of most mussel species can be divided into three parts:
proximal and distal regions and an adhesive plaque. The proximal region has a
corrugated surface and represents approximately one third of the total thread
length, whereas the distal region is smoother, narrower and approximately
twice the length of the proximal region. These two regions have been identified
in byssus from M. califonianus, M. edulis, M. trossulus and M. galloprovin-
cialis.3 Some mussel species show no differences between the proximal and
distal region. As an example, byssal threads from the South American mussel
Aulacomya ater show no morphological differences along the entire thread of
the byssus. In addition, problems have been reported when trying to char-
acterize the two regions of the thread of Bathymodiolus thermophilus, Geukensia
demissa and Dreissena polymorpha.3
Byssal threads have a fibrous core covered by a thin protective cuticle or
sheath. The thickness of this cuticle is in the range 2–4 mm.36 The cuticular
proteins show highly repetitive sequences, dominated by proline/hydroxypro-
line, lysine and 3,4-dihydroxyphenyl-L-alanine (DOPA), and mussel foot pro-
tein-1 (mfp-1).37,38 The function of such a cuticle is to protect the underlying
collagen fibres from abrasion and microbial attack.36
310 Chapter 14
Figure 14.3 SEM micrograph of the external morphology of mussel byssus from
Aulacomya ater.
14.3.3 Collagens Present in Mussel Byssus

Collagen is the major component of mussel byssus. The type of collagen that is
predominant in the distal region of byssus is known as Col-D. Col-D is a
homotrimer with a chain mass of 60 kDa. Col-P is present in the proximal
region of the thread and has an apparent mass of 55 kDa.28
Mussel foot extracts have been used to identify precursors of Col-P and
Col-D. The precursors are named preCol-P, preCol-D and preCol-NG. These
precursors have three major domains: a tough central collagen domain, flanked
on either side by structural domains. These structural domains resemble elastin
in preCol-P, spider dragline silk in preCol-D, and Gly-rich cell-wall proteins in
preCol-NG.15 PreCol-P is abundant in the elastic proximal region, while pre-
Col-D predominates in the stiffer distal portion. By contrast, PreCol-NG is
evenly distributed.
There are histidine-rich sequences in the terminal regions of the preCols in
mussel byssus. Histidines are proposed to be reversible ligands for coordination
with transition metal ions such as zinc(II) and copper(II). Such reversibly
broken coordination complexes are stronger than non-covalent bonds, but
possess only half the strength of a covalent bond.39 In addition, histidines are
suspected to play an integral role as sacrificial bonds in yield and self-healing in
the distal region of byssal threads.40
14.4 Characterization
14.4.1 Structural Characterization
The byssus fibre is composed of a fibrous core surrounded by a protective
cuticle. The structural morphology varies from species to species. The cuticle
of P. canaliculus is uniform and homogeneous. In contrast, the cuticle of
M. galloprovincialis has been described as a composite-like material having
granules immersed in a homogeneous protein matrix. These granules have a
diameter of around 0.8 mm and comprise about 50% of the cuticle volume.
In the case of M. galloprovincialis, the granular cuticle exhibits hardness and
stiffness approximately one order of magnitude greater than those of the
fibrous core. Holten-Andersen et al.41 have shown that this cuticle is extensible
(breaking strain B70%). They found that during the stretching of byssal
threads, microcracks were formed within the matrix of the cuticle. These cracks
did not propagate through the entire cuticle owing to the fact that the granules
arrest microcracks so that they cannot develop into microtears. Byssus cuticle is
the only known coating that has both high compliance and hardness which co-
exist without mutual detriment.42
Structural characterization of the adhesive plaque has been made in order to
understand the underlying principles of the attachment of mussels to hard and
soft surfaces. Byssal threads mediate attachment of the soft mushy body of the
mussel to a very stiff and hard substratum through their adhesive plaques.
When the adhesive plaque is joined to a hard substrate such as a rock, there is a
significant mismatch in the stiffness of the rock and the stiffness of the mussel
body. In order to prevent an excessive deformation of mussel body and mussel
byssus, the adhesive proteins within the adhesive plaque are present as solid
foam.43,44
The structure of the adhesive plaque from M. californianus is shown in
Figure 14.4. It is a continuous, partially open cell network in which the pore
size gradually increases from the interface towards the cuticle. The microfibrils
descending from the thread extend into the adhesive plaque. The damage
caused by contact deformation is mitigated by the pore size gradient within the
adhesive plaque.
The external morphology of plaques of A. ater is shown in Figures 14.4a and
14.4b. The shape of the plaques is elliptical. The patterned surface of plaques
plays an important role in the interaction with a variety of both organic and
inorganic substrates in aqueous environments. Through these plaques, mussels
secret adhesive proteins that allow them to attach themselves to hard sub-
strates. The adhesive proteins are characterized by the presence of an unusual
amino acid known as DOPA (3,4-dihydroxy-L-phenylalanine) that is thought
to be responsible for the strong adhesion of byssal threads.25,39,45
The morphological characteristics of assembled PreCols have been explored
by both transmission electron microscopy (TEM) and atomic force microscopy
(AFM).40,46 TEM of both the liquid crystalline polymer secretor granules and
byssal threads reveal a smectic packing that consists of ‘‘6 þ 1’’ bundles; that is,
312 Chapter 14
(a)
(b)
Figure 14.4 (a) SEM micrograph of an Aulacomya ater byssal adhesive plaque;
(b) SEM micrograph showing a closer view of the morphology of an
A. ater byssal adhesive plaque.
seven trimers per unit with six hexagonally arranged on the outside and one on
the inside. These can be modelled in three dimensions as flower or banana
shapes. AFM images have detected the banana variety. AFM studies have also
confirmed the extensive smectic arrays of these banana or bent core bundles.40

Byssus fibres need to meet specific mechanical requirements of stiffness,
strength, toughness and extensibility in order to withstand water velocities of
over 10 m s1 and water acceleration of around 400 m s2 in the marine inter-
tidal zone.47,48 In addition, the actual values of the mechanical properties, such
as ultimate stress, ultimate strain and others, vary from species to species.14
Figure 14.5 shows some representative stress–strain curves of different byssal
fibres from several mussel species. The energy absorbed during the deformation
process is represented by the area below the tensile curves depicted in Figure
14.5. The representative curve that corresponds to A. ater indicates that this
material absorbs more energy during the deformation process than the other
types of byssal fibres.
The mechanical properties of mussel byssus are also influenced by environ-
mental conditions such as temperature, humidity, salinity, etc.5,23,24,49 For
instance, Moeser et al.5 observed an increase in the strength and extensibility of
Figure 14.5 Representative stress–strain curves of byssus from (a) Dreissena poly-
morpha,1 (b) Bathymodiolus thermophilus,1 (c) Modiolus modiolus,1
(d) Mytilus edulis,1 (e) Geukensia demissa1 and (f) Aulacomya ater.
314 Chapter 14
byssus fibres after fall and winter. They also report that the strongest attach-
ment occurs during spring, while the weakest attachment takes place in fall.
The distal and proximal regions of byssal fibres from several mussel species
have different mechanical properties among each other. In the case of M. edulis,
Allen et al.9 found that the distal region was stronger than the proximal region.
Smeathers and Vincent10 reported that the proximal region is more extensible
than the distal region. Waite et al.40,46 found that the distal region of the mussel
byssus from M. galloprovincialis is about 10 times stiffer than the proximal
region.
Bell and Gosline investigated the stress–strain behaviour of full threads and
individual proximal and distal regions of several mussel species.14 They showed
that the byssal threads of edulis-like species have both similar compositions
and mechanical behaviour. They found major differences between the distal
and proximal regions. Typical stress–strain curves for isolated distal and
proximal regions showed that the proximal region has a lower Young’s mod-
ulus, a lower ultimate stress and a higher ultimate strain than the distal region.
The distal region exhibits a striking yield behaviour (at around 35 MPa) and is
considerably stronger than the proximal region.
In the case of entire byssal fibres, the same authors divided the stress–strain
curve into three phases: the mechanical behaviour of the first phase is due to the
stretching of the proximal region; the second phase of the curve is dominated
by the yield of the distal region; whereas the third phase shows the stiffening of
the distal region.14
Carrington and Gosline have reported cyclical loading tests carried out on
byssal threads. The mechanical behaviour of mussel byssus depends on the
loading history of the thread. In Mytilus californianus, whole threads cycled
below the yield point were highly resilient and increased slightly in stiffness
when loaded once again. By contrast, threads cycled beyond the yield point had
much lower resilience and were dramatically less stiff when reloaded.3
Harrington and Waite have observed the behaviour of native seawater-
treated distal threads when cycled multiple times, to a slightly higher strain each
time. They noted a second yield point for cycles beyond the initial yield point.
After each successive yield, the threads showed a reduction in the elastic
modulus. The largest reduction in the stiffness occurs during the 20% strain
cycle, with the initial modulus reduced by almost 40% from the first cycle.15
Troncoso et al. have studied the mechanical properties of mussel byssus from
A. ater. They used true stress and strain to model the mechanical behaviour of
byssal threads in hydrated and dry environments. Linear, power law-type and
Mooney–Rivlin relationships were used to model the results obtained from
their uniaxial tensile tests.8
The results of the tensile tests carried out by Troncoso et al. are summarized
in Table 14.2.8 The mechanical properties of byssus immersed in distilled and
sea water are similar. By contrast, hydrated byssus is more extensible than dry
byssus, as it reaches higher maximum strain values. Furthermore, differences
between true and engineering ultimate stresses are similar for both dry and
hydrated byssus.
Table 14.2 Values of mechanical properties of mussel byssus from A. ater
under different conditions.
Condition Ultimate stress (MPa) Ultimate strain
Dry 144.71 33.58 0.27 0.018
Immersed in distilled water 59.47 25.02 0.84 0.097
Immersed in sea water 70.16 20.46 0.79 0.275
Figure 14.6 shows representative stress–strain curves of byssal threads. Dry

byssal thread (Figure 14.6a) behaves as an elastoplastic polymer while humid
threads (Figure 14.6b) immersed in both distilled and sea water display elas-
tomeric behaviour. Figure 14.6a shows that the curves of dry byssus are
characterized by two clearly different regions. Initially, they follow a linear
elastic behaviour at small strains until they reach a critical strain where plastic
deformation begins. A power-law equation has been used to describe such
plastic behaviour.8 The suitability of this power-law to model the tensile
behaviour of dried byssal threads is clear from Figure 14.6a, where a repre-
sentative stress–strain curve for dry byssus is shown.
Figure 14.6b shows a representative stress–strain curve for hydrated byssus,
demonstrating that the experimental data fit well with the chosen model. In
contrast to dry byssus, lower stresses are needed to deform the hydrated byssal
threads. Troncoso et al. used the Mooney–Rivlin relation of rubber elasticity
theory to model the stress–strain relationships, owing to the fact that hydrated
byssus displayed an elastomer-like tensile behaviour (theoretical points in
Figure 14.6b). The Mooney–Rivlin model has been used in the past to model
the deformation of rubber-like materials and has been applied to the study of
soft tissues such as tendons and ligaments.50–53
14.4.3 Thermal Properties

Differential scanning calorimetry (DSC), thermomechanical analysis (TMA)
and dynamic mechanical analysis (DMA) have been used to evaluate the
thermal properties of byssus fibres. Tsukada et al. used DSC to examine
thermal transitions of byssus fibres.54 They found a minor endothermic peak at
230 1C and a major broad endothermic peak at 310 1C. The last broad endo-
thermic peak was associated with the thermal decomposition of the collagenous
byssus fibres. Figure 14.7a shows DSC curves of byssus fibres. Dried byssus
fibres have a thermal transition at around 103 1C, whereas hydrated byssus
fibres show a thermal transition at around 113 1C. Although the temperature of
the thermal transitions of hydrated byssus fibres is different from the thermal
transition of dried fibres, when the amount of water content rises, the thermal
transition temperature does not change accordingly.
TMA tests showed that the byssus fibres slightly contracted at a constant rate
during heating from room temperature to about 200 1C. After that, the con-
traction increased abruptly, reaching a maximum value at 240 1C. Beyond this
316 Chapter 14
(a)
(b)
Figure 14.6 (a) Experimental () and theoretical (–) stress–strain curve for dry byssus
from A. ater; (b) experimental () and theoretical (–) stress–strain curve
for hydrated byssal threads from A. ater immersed in sea water.
temperature, the byssus fibres started to expand rapidly. The latter might be
attributed to the fast thermal movement of the molecules, along with the
decomposition and partial reforming of the intermolecular bonds. These results
suggested that both the high degree of contraction registered by the TMA test
above 200 1C and the minor endothermic peak (around 230 1C) are related to
the occurrence of some changes in the byssus fibres at a molecular level.54
Aldred et al. have used DMA tests to study mussel byssus fibres (Figure
14.7b).55 They found that the denaturation and thermal decomposition of
(a)
(b)
Figure 14.7 (a) Representative thermograms of mussel byssus with different water
contents; (b) dynamical mechanical analysis of mussel byssus in dry
conditions.55
byssus fibres start at 217 1C. They determined that the glass transition tem-
perature (Tg) of dried byssus fibres takes place at around 6 1C.
14.4.4 IR Analysis
IR analyses of peptides and proteins are characterized by typical absorptions
peaks such as amide bands. Figure 14.8 shows the IR spectra of byssus fibres
from A. ater. These byssus fibres exhibit an amide I band showing a narrow
peak at B1655 cm1. Amide I (B1600–1700 cm1) is associated with the C¼O
318 Chapter 14
Figure 14.8 FTIR spectra of a byssal thread from A. ater.
stretching vibration (70–85%) and the C–N stretching vibration (10–20%).56

Different species show similar results but not identical values. Hagenau et al.
studied the species M. galloprovincialis, separating distal and proximal
regions.57 Their results showed that the amide I peak of the distal region occurs
at 1633 cm1, whereas the amide I peak of the proximal region takes place at
1626 cm1. The differences on the exact position of this band are determined by
the backbone conformation and the hydrogen bonding pattern.56
Byssus fibres also show amide II and III peaks at B1555 cm1 and
B1235 cm1, respectively (Figure 14.8). The amide II peak (B1520–1580 cm1)
is associated with the N–H bending vibration (40–60%), C–N stretching
vibration (18–40%) and the C–C (B10%) stretching vibrations, whereas the
amide III peak (1229–1301 cm1) is related to the N–H bending and C–N
stretching vibrations within the triple helical structure of collagen.56 Hagenau
et al. have reported the amide II peak for the distal and proximal regions at
1530 cm1 and 1543 cm1, respectively.57
Figure 14.8 also shows a broad peak at 3300 cm1. This peak is associated with
N–H stretching vibrations that give rise to amide A (3310–3270 cm1).58 The
narrow signals at around 2900 cm1 correspond to C–H stretching vibrations;
C–H deformation vibrations appear at around 1450 cm1. The IR signals
between 1200 and 1300 cm1 are typically assigned to collagen types I and II.57
Kong et al. have used FTIR curves to perform a quantitative estimation of
the content of different secondary structures.56 They used a curve fitting pro-
cedure to analyze the deconvoluted spectrum of the amide I band. They argue
that any protein can be considered as a linear sum of a few fundamental sec-
ondary structural elements and that the percentage of each element is only
related to the spectral intensity. In order to study the secondary structure of
byssus fibres from A. ater, the amide I band was deconvoluted, and the
Table 14.3 Percentage of each secondary structure of byssus fibres from
A. ater.
Secondary structure Wavelength (cm1) Percentage (%)
a-Helix 1657 22.5
b-Sheet 1622, 1639 36.8
b-Turn 1676 17.8
deconvoluted spectrum was be fitted with Gaussian band shapes. The decon-
voluted amide I band frequencies, assignments to secondary structure for
byssus fibres of A. ater, as well as the percentage of their secondary structures,
are shown in Table 14.3. The results confirm the presence of crystalline and
amorphous regions in the secondary structure of byssus fibres with a high
b-sheet (B37%), a-helix (23%) and b-turn (18%) structural content.
Hagenau et al.57 studied the secondary structure of byssus fibres of
M. galloprovincialis. They found that for byssus fibres which can be divided
into two parts, distal and proximal regions, they present secondary structure in
the following way. Proteins of the distal region are oriented along the thread
axis and are well ordered with a high b structural content (70%) and collagen
typical triple helices (22%). Proteins of the proximal part are not as well
oriented and show less order, with primarily a-helical structure (47%) with less
b conformations (15%) and triple helical structures (13%).
14.4.5 Adhesive Properties

The natural adhesive used by mussels is composed of glue proteins that are able
to bind strongly to virtually all inorganic and organic surfaces in aqueous
environments in which most adhesives work poorly.39 It is known that byssal
threads counteract the residual stresses that arise at the interface between the
substratum and the adhesive plaque of mussels by the production of a struc-
tural adhesive that appears in the form of solid foam.43,59–61 Several theories
have been developed to explain the process by which this foam is made, but it is
still unclear.
The adhesive plaque is composed of six distinct types of proteins known
as foot proteins (fp) (Table 14.4).30–34,62–64 Even though M. edulis has been
the main mussel species used for identifying foot proteins, studies carried out
on other mussel species, such as M. galloprovincialis, M. coruscus and M.
californianus, confirm that they have similar amino acid sequences of adhesive
proteins.25,65,66
Studies reveal that the mussel adhesive proteins of M. edulis contain a high
level of the amino acid DOPA. DOPA is a catabolic amino acid that is pro-
duced by post-translational hydroxylation of tyrosine using polyphenol oxi-
dase.31,32,34 DOPA-containing adhesive proteins are also found in other
mussels and even in other phyla as well. It was reported that mussel adhesive
proteins which are closer to the adhesion interface have a higher proportion of
320 Chapter 14
Table 14.4 Mass (kDa) and DOPA content (mol%) of mussel byssus foot
proteins.
Proteins Mass (kDa) DOPA (mol%) Ref.
Mefp-1 B110 B10–15 33,64
Mefp-2 B42–47 B2–3 73
Mefp-3 B5–7 B20–25 31,64,73
Mefp-4 B79–80 B4–5 31,73,74
Mefp-5 B9 B27–30 73,77
Mefp-6 B12 B4 77
DOPA residues, whereas mussel adhesive proteins with less or without DOPA
show a reduced ability for adhesion.31,33,34 In addition, DOPA residues allow
mussel adhesive protein molecules to cross-link each other by oxidative con-
version to DOPA-quinone. It has been suggested that the reactive quinone
provides the water resistance characteristic of mussel adhesion.67,68 Therefore,
DOPA functions as a cross-linking agent and mediates adhesion to the
substratum.
The first mussel adhesive protein to be identified was fp-1 from M. edulis
(Mefp-1).33,69 This protein is found on the thin protective cuticle that covers the
entire structure of the adhesion plaque and the distal region of the thread.
Mefp-1 has a high molecular mass as it is composed of about 80 repeats of a
decapeptide (Ala-Lys-Pro-Ser-Tyr-Hyp-Hyp-Tyr-DOPA-Lys). The DOPA
content is 10–15 mol% and has the highest molecular mass among mussel
adhesive proteins (110 kDa). Mefp-1 is directly related to the durability of byssus
fibres and its robust coating is 4–5 times stiffer and harder than the byssal
collagens that it covers.42 Analogous proteins with slight differences in the
amino acid composition have been reported to be present in M. galloprovincialis
(Mgfp-1), M. coruscus (Mcfp-1) and M. californianus (Mcfp-1).70–72
Mefp-2 and Mefp-4 are located in the bulk of the mussel’s adhesion plaque.
Unlike Mefp-1, Mefp-2 is a smaller adhesive protein (40–47 kDa) with
approximately 2–3 mol% of DOPA.64,73 Mefp-2 contains abundant secondary
structure and is relatively resistant to a variety of proteases, which is an
important characteristic for the integrity of the byssal plaque. In addition,
Mefp-2 is an adhesive protein that plays a stabilization role due to its high
cysteine content (6–7 mol%).30
Mefp-4 has a molecular mass of B80 kDa and a DOPA content of
4 mol%.64,74,75 Mefp-4 also contains high levels of arginine, histidine and gly-
cine.64,73 This protein likely serves as a coupling agent in the thread plaque
junction designated by the pre-collagens and the byssal plaque protein Mefp-2.75
The interface between the substratum and the adhesion plaque of mussels is
formed by Mefp-3 and Mefp-5 proteins. Both proteins contain higher levels of
DOPA. Mefp-3 contains 20–25 mol% DOPA, whereas Mefp-5 contain about
30 mol% DOPA.73
Mefp-3 is the smallest adhesive protein (B5–7 kDa), with a large number of
arginine residues.31 Other mussels contain analogous proteins. Mgfp-3A and
Mgfp-3B have been identified in the adhesive plaque of M. galloprovincialis,
whereas Mcfp-3 (12 variants) have been found in M. californianus.76,77
Mefp-5 is a relatively small protein (9.5 kDa). It has many serine residues
that can partly be modified to phosphoserines. A protein analogous to Mefp-5
has been reported to be present in M. galloprovincialis (Mgfp-5)65 and in
M. californianus (Mcfp-5: 2 variants).63
Mcfp-6 was identified along with Mcfp-5 by Zhao and Waite.63 This small
protein (11.6 kDa) contains a relatively large amount of tyrosine (B20 mol%)
and a small amount of DOPA (o5 mol%). It is suggested that its function is to
provide a cohesive link between the surface-coupling DOPA-rich proteins and
the bulk of the plaque proteins.
14.5 Potential Applications and Biomimetics

14.5.1 Biomimetics
Some characteristics of mussels have attracted the interest of researchers,
especially in the field of biomimetics. Mussels are able to produce their byssus
fibres and plaques in a few minutes, and these fibres spontaneously adhere to
rocks. The adhesive process can be described as follows. The concentrated
adhesive proteins, which are stored in a Ca21/Mg21 rich medium at pH 5, are
found in the secretory granules located in the cement gland. The secretory
granules await a signal transduction to release their contents. When mussels
want to attach themselves to hard surfaces, proteins are released onto the
surface, flowing rapidly. It has been suggested that the condensed state of the
proteins helps to dehydrate the hard surface.43 Finally, the DOPA residues will
be oxidized to quinones, producing a cross-linking process that leads to
solidification.
One of the most interesting techniques that mussels use is the fast bonding
process to metal and mineral surfaces in a wet saline environment at ambient
temperature. This behaviour can be observed in other marine organisms, such
as barnacles, tubeworms, sea cucumbers and limpets. This natural adhesive
process has not been imitated by synthetic polymers since they cannot displace
surface water. The effect of moisture affects the stability of interfacial adhesion
between metal and polymer composites.
Waite et al. indicated that at least 10 proteins are present in the adhesive
plaques, but only two of these, Mefp-3 and Mefp-5, are implicated as inter-
facial.43 Both are among the most modified of all byssus proteins, with 42%
and 37% of modified amino acids, respectively. The main modification in both
proteins is tyrosine hydroxylation to DOPA. It has been shown that the
o-dihydroxyphenolic moiety of DOPA is implicated in the strong H-bonding
to hydroxyapatite and coordination complexes with the oxides of iron,78 zinc,79
titanium,80 aluminum81 and gold surfaces.82
The different DOPA-containing precursors are stockpiled in specific places in
the byssus fibres. In the cuticle, DOPA mediates formation of covalent
322 Chapter 14
III
diDOPA cross-links of bis- and triscatecholato-Fe complexes with
Mefp-1,83–85 and even triscatecholato-FeIII complexes that serve as vehicles for
diDOPA cross-link formation by a one-electron redox exchange.86 Mefp-3 and
Mefp-5 are deposited at the interface with a hard surface and remain with their
DOPA intact (surface metal chelates).43
Mussel byssus is a source of inspiration for the design of novel biomimetic
polymers and composites. Waite suggests that mussel adhesive is essentially
analogous to composite thermosets. It consists of fibres that are dispersed in a
cross-linked resin, rendering it resistant to heat and solvents. The fibres are
formed by collagen and the resin is proteins with basic isoelectric points, high
levels of the amino acid DOPA and an extended, flexible conformation.25
One of the attributes why mussel adhesion has attracted interest for potential
applications is that mussels can adhere themselves to a variety of substrates.43
The adhesion of these glue proteins to various solid materials, such as plastic,
glass, metals, Teflon and even living body substances such as porcine skin and
diverse types of mammalian cells, has been verified.62,64,65,87–93 Another
advantage of mussel byssal adhesion is that it is fully functional under water or
in humid conditions.4 All stages of the bonding process occur rapidly under
ambient and wet conditions. Finally, byssal adhesion appears to be a smart
process in the sense that it exhibits compliance or modulus matching.40 In
addition, byssal threads are environmentally friendly and biodegradable
materials. Therefore, mussel adhesion has a wide range of potential uses in
diverse biotechnological applications.
14.5.2 Surgical Adhesives

Ideal surgical adhesives should adhere to the tissue substrate, providing ade-
quate strength in the presence of physiological fluids. They should also be
biodegradable and should not elicit an immune response.93 Ninan et al.
examined the adhesive characteristics of glue proteins extracted from mussels
without the use of a curing agent.93 These glue proteins were used to bond
porcine skin in an end-to-end joint cured in controlled environments similar to
common surgical environments: dry and humid environments such as those
used for external and internal incision. When comparing the results with those
obtained from the commercially adhesive fibrin Tisseelt, they concluded that
joints bonded with mussel extract showed adhesive strengths similar to those
bonded with fibrin for cures times between 12 and 24 hours. This extract can be
used as a potential biomaterial for surgical applications if the rate of cure can
be accelerated and show biocompatibility properties as well.
14.5.3 Dental Adhesives

Mussel adhesives can be used as medical and dental adhesives as they are
harmless to the human body and do not impose immunogenicity.94,95 Tay and
Pashley suggested that future medical and dental adhesives may contain
domains derived from protein-based, underwater bioadhesives secreted by
aquatic animals such as mussels, making them less dependent on the surface
energy of the bonding substrate and less susceptible to hydrolytic degrada-
tion.93,96–99
Because DOPA residues are known for their ability to impart adhesive
and curing properties to mussel adhesive proteins, hybrid systems containing
Mefp-1 decapeptides and synthetic polymers have been used for the production
of hydrogels for medical applications. Lee et al. prepared hybrid systems
containing linear and branched DOPA-modified poly(ethylene glycol)s (PEG
DOPAs), resulting in improved adhesive and cohesive properties.100 This gel
can potentially function as a bioadhesive material for medical purposes. Other
hybrid systems that are made of Mefp-based peptides and synthetic polymers
such as different acrylates have been developed at the Fraunhofer Institute for
Manufacturing Technology and Advanced Materials by solid-phase peptide
synthesis for dental applications. These hybrid systems showed improved ten-
sile shear strength compared with pure acrylate systems.
14.5.4 Biosensors and Immunosensors

Biosensors combine the molecular recognition capabilities of biology with the
signal processing capabilities of electronic devices. In order to fabricate bio-
sensors, adhesives are needed to integrate biological components into micro-
fabricated devices. The immobilization of mussel adhesive proteins such as the
Mefp class and collagens on solid supports may be exploited for uses in the
design of biosensors, immunosensors or artificial tissue scaffolding and con-
structs.101,102 Saby and Luong constructed a glucose oxidase (GOD) based
electrode using an adhesive protein isolated from M. edulis.101 Then the mussel
adhesive protein was oxidized to form a stable film on platinum, gold or glassy
carbon (GC) electrodes. They were covalently linked to the GOD. The modified
electrodes were able to detect tetrachlorobenzo-1,4-quinone (TCBQ) under
specific conditions. In the case of GC/mussel adhesive protein/GOD electrodes,
they exhibited a linear response for TCBQ, ranging from 10 nM to 1 mM (3.96
nA nM1) with a detection limit of 10 nM, retaining about 90% and 87% of its
activity after 150 and 250 repeated injections, respectively. Yamada et al.
suggest that the identification and subsequent use of the polyphenol oxidase
derived from M. edulis byssal structures may also be exploited as a thickening
agent for numerous industries and applications.103 Hwang et al. prepared
modified recombinant Mgfp-5 that can be used as a novel immobilization agent
for biochip systems or as a mediator for drug delivery.88
14.5.5 Cell Culture and Tissue Engineering

Several cell-adhesion materials have been developed in order to coat artificial
surfaces and promote cell adhesion. Among them are extracellular matrix
proteins, poly-L-lysine (PLL), Cell-Tak and mussel adhesive protein (MAP).
324 Chapter 14
Extracellular matrix proteins have been widely used in cell culture and tissue
engineering because they have various cell-adhesion motifs. However, they are
expensive and are associated with infection risks since they are extracted from
foreign animals. By contrast, PLL is cheaper than extracellular matrix proteins
but it is not always an effective cell-adhesion material due to cytotoxicity and
abnormal cell spreading.104
A commercially available cell-adhesive agent based on mussel adhesive
proteins is Cell-Tak (BD Bioscience Clontech). It is an extracted mixture
derived from Mefp-1 and Mefp-2. Cell-Tak has less cytotoxicity and promotes
improved cell-adhesion ability among various cell types, including mammalian
and human cell lines.105 Although Mefp-1 and Mefp-2 have no higher DOPA
levels than Mefp-3 and Mefp-5, these products have been used to improve the
attachment of cultured cells and tissues. It is now quite evident that the role of
Mefp-1 is related mainly to its coating function rather than with its adhesion
properties, owing to its lower DOPA levels.42 Even though mussel adhesive
proteins have many remarkable properties, their practical applications are
restricted by an inefficient generation process and an extremely low production
rate.106
In order to overcome these disadvantages, Hwang et al. designed and con-
structed a hybrid of the MAP fp-151, which is a fusion protein with six type
1 (fp-1) decapeptide repeats at each type 5 (fp-5) terminus.89 It was found that
fp-151 has advantages such as high production yield in Escherichia coli and
simple purification. Several cell-adhesion experiments have demonstrated the
potential use of fp-151 for cell or tissue adhesion.
Hwang et al. have designed a new cell-adhesive protein called fp-151-RGD
that is a fusion of the Arg-Gly-Asp motif to hybrid MAP fp-151.91 The Arg-
Gly-Asp (RGD) sequence is the most effective cell-adhesion recognition motif
that can be found in materials, such as collagen, fibronectin and tenascin C, and
has been used to stimulate cell adhesion on artificial surfaces.107–110 The results
showed that fp-151-RGD had the advantages of fp-151, but also showed
superior cell adhesion and spreading abilities under serum-free conditions
regardless of mammalian cell type compared with other commercially produced
cell-adhesion materials such as poly-L-lysine (PLL) and the naturally extracted
MAP mixture Cell-Tak. These properties might be explained due to three cell-
binding mechanisms: cationic binding, DOPA adhesion and the cell-adhesion
motif. The results demonstrated the suitability of fp-151-RGD as a cell-adhe-
sion material in cell culture and tissue engineering. In addition, fp-151 has the
potential to be fused with other specific motifs instead of RGD motifs, so that
specific target cells can be tightly bonded to an artificial extracellular matrix or
biomaterial; thus fp-151-based fusion proteins with specific cell-recognition
motifs can be used in tissue engineering as specific cell-adhesion biomaterials.
Proteins with a high DOPA content have been studied as cell-adhesion
biomaterials. Hwang et al. reported the production of recombinant M. gallo-
provincialis foot protein type-5 (Mgfp-5) in E. coli,65 showing that the
recombinant protein had better adhesive properties than Cell-Tak in terms of
surface adhesion and cell immobilization. They also used purified and
tyrosinase-modified recombinant Mgfp-5 to adhere living anchorage-
independent cells such as insect drosophila S2 cells and human MOLT-4 cells
onto glass slides, finding that the modified recombinant Mgfp-5 can be used as
an adhesive biomaterial for cells.
14.5.6 Biotechnological Applications

Efficient target gene delivery into eukaryotic cells is a key issue to gene therapy
and biotechnological research. In this field, various histone proteins have been
analyzed as potential gene delivery materials because they display higher
transfection efficiency in mammalian cells compared to the widely used
transfection agent Lipofectamine 2000. In order to find new alternatives, a
recombinant mussel adhesive protein has been assessed as a potential gene
delivery material. Hwang et al. investigated the use of the recombinant mussel
adhesive protein fp-151 as a gene delivery material,90 because hybrid fp-151
displays a similar basic amino acid content and high theoretical pI values as
histone HI proteins, and exhibits efficient DNA binding ability. Hwang et al.
transfected mammalian cells (human 293T and mouse NIH/3T3) with foreign
genes using the hybrid fp-151 as the gene delivery carrier. The results showed
that this hybrid protein displays a transfection efficiency comparable with the
efficiency of the widely used Lipofectamine 2000. Therefore, mussel adhesive
protein may also be used as a potential protein-based mediator for efficient
gene delivery.
14.6 Conclusions
In this chapter we have reviewed some important characteristics and properties
of the mussel byssus. The mussel byssus is a tough biopolymer fibre that occurs
in nature and shows remarkable mechanical as well as adhesive properties. The
composition of the proteins that allow mussel byssus fibres to adhere them-
selves to many different substrates in dry and humid environments has been
reviewed and an up-to-date summary of the main applications that are inspired
by or take advantage of these adhesive proteins have been reported. These
applications include surgical and dental adhesives, biosensors and adhesives for
tissue engineering.
The mechanical properties of byssal fibres have also been reviewed, since
their toughness is six times greater than that of the human tendon collagen and
is comparable with that of Kevlar. It has been shown that the mechanical
behaviour of such fibres is influenced by environmental conditions such as
temperature, humidity and salinity, among others. For instance, dry byssal
fibres show an elastoplastic stress–strain curve, while humid byssus display an
elastomeric-like behaviour. As in the case of other protein-based fibres such as
spider silk and silkworm silk, the understanding of the underlying principles
that allow such remarkable mechanical behaviour is important for the devel-
opment of new applications for mussel byssus.
326 Chapter 14
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Subject Index
The index covers both volumes. The volume number is given in bold
before the page numbers. Page numbers in italics refer to figures or tables.
abaca (Musa textilis) Agave sisalana see sisal

composition 1:43, 90, 126 agricultural bioplastics 2:107–108
production and cost 1:39, 120 alginate 2:251
properties 1:40, 41, 42, 46, 88 alkali treatment 1:53, 235, 2:13
abaca fibre composites 1:92, 129 1-allyl-3-methylimidazolium chloride
Abelmoschus esculentus see okra ([amim]Cl) 2:195–197
ABS see acrylonitrile/butadiene/ alumina
styrene in artificial nacres 2:142–143
absorption see water sorption of porous membrane
natural fibres templates 2:161–163
Abutilon theophrasti see velvet leaf alumina hydrate see aluminium
acetylation of wood 1:155–156 hydroxide
acidic hydrolysis of cellulose 2:18–19, alumina trihydrate (ATH) see
22–23 aluminium hydroxide
acoustic insulation 1:40, 55 aluminium, infiltration into spider
acrylic acid 2:46 silk 1:283
acrylic resin composites 1:90, 104–106 aluminium hydroxide (aluminium
acrylonitrile/butadiene/styrene (tri)hydrate)
(ABS) 1:93, 231, 235 as flame retardant 1:106–107
adhesion, interfacial see interfacial in soy protein
interactions nanocomposites 2:99–100
adhesion, work of 1:184, 197–198, aluminium Keggin ions,
198 intercalated 2:47–48
adhesion agent, poly(furfuryl alcohol) [amim]Cl see 1-allyl-3-
(PFA) as 2:166 methylimidazolium chloride
adhesives amino acids, in silk 1:25–26, 221,
medical and dental 1:322–323, 276–277
2:278–279, 280 ammonium polyphosphate
wood 2:103–104 (APP) 1:107–110
aerospace applications 1:100 amylopectin 2:186, 187–189, 190–191
Agave fourcroydes see henequen see also waxy maize starch
Subject Index 331
amylose 2:186–187, 189–191, 190, silk-based
213–214 composites 1:231–232,
amylose starch/nanoclay composites 236–239, 238, 240, 240–242,
effect of plasticizers 2:69–70, 243, 244–249
72–89 soy protein
chemical interactions 2:75–80 nanocomposites 2:103–108
moisture content 2:81–84, 86–87 starch nanocomposites 2:230
physical interactions 2:70–75 zein 1:213–214
synergistic interactions 2:84–89 aprismatic enamel see prismless
Ananas comosus see pineapple enamel
Ananas erectifolius see curaua aragonite 1:18–20, 22–23, 2:97–99,
angiogenesis properties 1:236, 2:272, 114–115
278 aramid fibres 1:87, 88
animal fibres 1:6, 66, 125 artichoke 1:40, 41, 42, 43, 46, 55
see also mussel byssus fibres; silk; artificial nacres 1:29–30, 2:48, 51–52,
spider silk; wool 141–145
anisotropy asbestos 1:66
in nacre 2:116 aspect ratios 1:66
in natural fibres 1:64, 73–75, 77–80 cellulose nanocrystals 2:21, 22, 24
antimicrobial nanocomposites 2:45, nacre tablets 2:117, 118, 119–120
58–59, 250 nanofillers 2:37–38
apatite see hydroxyapatite natural fibres 1:39, 40
(HA, HAp) atomic force microscopy
applications 2:247–251 (AFM) 1:185, 189
see also biomedical applications; Aulacomya ater 1:309, 311–312, 313,
tissue engineering 314–315, 315, 317–319, 319
adhesives, wood 2:103–104 autologous cell therapy 2:283
automotive industry 1:80–81, 87, automotive industry 1:118–137
129–137 advantages of natural
biosensors 1:248–249, 296, 323, materials 1:37–38, 52–53, 81,
2:35, 37, 39, 214 118–121
biosorbents 2:58, 60 application of natural fibres 1:87,
chitosan-based 2:35, 36–37, 59–61, 129–135, 136
250 flammability performance
immobilization (enzymes, requirements 1:99
etc.) 1:248–249, 295–296, 323, polymer/layered silicate
2:35, 39 nanocomposites 2:247–248
mussel byssus fibres 1:321–325 Avrami equation 2:78–80, 81
natural fibre composites 1:80–81,
87, 129–137 bacterial cellulose 2:21, 202–203
packaging materials 2:59, ball milling 2:237
106–107, 248–250 bamboo
silk cellulose crystals from 2:203–205
raw and spun 1:222–223 composition 1:43
regenerated silk-based properties 1:40, 41, 42, 43, 46
materials 1:226 bamboo composites 1:164, 172, 2:102
332 Subject Index
banana biomimetics 1:29–31, 2:250–251
cellulose nanocrystals from 2:21 artificial nacre 1:29–30, 2:48,
composition 1:43 51–52, 141–145
properties 1:40, 41, 42, 46 hybrid soy protein
banana fibre composites 1:76, 92, 164 nanocomposites 2:97–100
bark of cotton stalk 1:40, 42, 43, 46 biopolymers, definition and
barrier properties 2:55–57, 248, 249, 250 examples 1:2–3, 4, 164, 165,
soy protein-based films 2:106–107 260–261
starch composites 2:208, 223, 224, bioregenerative engineering see tissue
228 engineering
zein films 1:211, 212 bio-resins (defined) 1:135
basal spacing, in nano clay 2:72, 86 biosensors 1:248–249, 296, 323, 2:35,
benzyl(dodecyl)dimethylammonium 37, 39, 214
bromide (12-OREC)-activated biosorbents 2:58, 60
montmorillonite biotechnological applications 1:325
(OMMT) 2:199–202 birefringence, in cellulose
biocomposites (defined) 1:229, 261–262 nanocrystals 2:22–23
biodegradability blood coagulation 2:260, 261,
agricultural bioplastics 2:107–108 263, 265
FA/PFA-modified sisal blood plasma 2:258–259, 263
fibres 2:170–171 Boehmeria nivea see ramie
packaging materials 2:59 bone
polymers 1:94, 164, 165, 260–261 hierarchical structure 1:8–14, 30
starch nanocomposites 2:195, 229 tissue engineering 2:250–251, 268,
biomass wastes 1:257–260, 261, 2:10 279–280
biomedical applications 2:250–251 silk composites in 1:241–242,
see also tissue engineering 243, 244, 247, 248
adhesives 1:322–323, 2:278–279, 280 bone matrix gelatin 2:279
chitosan-based 2:35, 36–37, 60–61, building industry 1:80, 81, 87
250 bundles, fibre 1:47–48
collagen-based 1:263–264, burn treatment 2:268
2:260–261, 267–270 butadiene rubber composites 1:263
drug delivery systems 1:246, byssus fibres see mussel byssus fibres
2:60–61, 262, 271, 276, 278
elastin nanocomposites 2:273–278 cadmium sulfide (CdS)
fibrin nanocomposites 2:278–280 nanoparticles 1:239
gelatin nanocomposites 2:271–273 calcite 1:22–23, 2:97–98, 104
silk calcium carbonate (CaCO3) 1:124
raw and spun 1:223 aragonite 1:18–20, 22–23, 2:97–99,
regenerated silk-based 114–115
materials 1:226 calcite 1:22–23, 2:97–98, 104
silk-based composites calcium silicate (CaSiO3) 1:243,
1:235–236, 237, 238, 240, 244–245
240–242, 243, 244–245, cancellous bone 1:13
246–247 Cannabis sativa see hemp
spider silk 1:294–296 carbamide, as plasticizer 2:199–200
Subject Index 333
carbon fibres 1:87, 88 thermal stability and
carbon molecular sieve (CMS) flammability 1:89, 2:14, 23
membranes 2:159–163 thin film preparation 1:186
carbon nanotube (CNT)-coated cellulose, carboxymethyl 2:13,
fibres 1:234 219–220
carbon nanotube (CNT) cellulose, 2-hydroxyethyl
composites 2:246–247 (HEC) 1:264–266, 267–270
with carbon 2:164 cellulose acetate butyrate
with chitosan 2:37, 39, 48, 51 (CAB) 2:171–172
with PFA 2:163–169 cellulose diacetate (CDA) 1:51
with silk 1:248 cellulose fibre composites 1:92, 164,
with soy protein 2:96–97 165
with starch 2:210–215, 216, 247 with coupling agents 1:171, 173
carbon/carbon nanocomposites effects of fibre orientation 1:79–80
2:164 with gelatin 2:271–272
carbons, porous 2:154–159 with starch 1:69–70
carboxymethyl cellulose 2:13, cellulose microfibrils 1:41–43, 66,
219–220 2:3–4
cardanol derivative of toluene see also microfibrillated cellulose
diisocyanate (CTDIC) 1:171–172 cellulose nanocrystal (whisker,
cartilage 2:278, 279, 280 nanowhisker, (CNW)) composites
cassava bagasse 2:21 with PFA 2:173–177
cassava starch 2:194–195, 207–208 with silk 1:245
Ceiba pentranda see kapok with soy protein 2:100–101
celery 1:40, 41, 42 with starch 2:203–205
cell adhesion 2:58, 265–266, 267, cellulose nanocrystals (whiskers,
272–273, 280–281, 283, 284 nanowhiskers (CNW)) 1:6–7,
cell adhesion agents 1:323–325 2:18–25
cell behaviour with see also cellulose nanocrystal
nanostructures 2:283–285 composites
cell migration 2:272, 275, 281, 283, aqueous suspension 2:22–23
284 films 1:186, 2:23–25
cell sources for tissue mechanical properties 2:4–6, 7–8,
engineering 2:278, 281–283 23–25
cellulose 1:3 morphology 2:19–22, 21
see also cellulose fibre composites; preparation 2:18–19
cellulose nanocrystals; cellulose nanofibrils 2:3–4
microfibrillated cellulose see also microfibrillated cellulose
acidic hydrolysis 2:18–19, 22–23 cellulose nanoparticles 2:2–8,
adhesion forces 1:192–194 202–205
natural fibres composition 1:90, see also cellulose nanocrystals;
125, 126 microfibrillated cellulose
PFA-modified 2:171 cellulose nanowhiskers (CNWs) see
properties 1:40, 41, 2:4–8, 7 cellulose nanocrystals
structure 1:41–43, 65–66, 74–75, cetyltrimethylammonium bromide
2:2–5 (CTAB)-modified MMT 2:46
334 Subject Index
China reed see Miscanthus coir (coconut fibre) composites 1:92,
chitin 1:3, 2:34–35 163, 231
as coupling agent 1:172–173 applications 1:129, 131
in seashells 1:21 flame retardation 1:112
whiskers 1:246, 2:100 furfuryl alcohol treatment 1:174
chitosan (CS) 1:3, 7, 2:34–36 collagen 1:3, 263–264, 2:259–261
as coupling agent 1:172–173 in bone 1:9–11, 13–14
chitosan-based isolation of 1:262, 2:257
(nano)composites 2:36–38, 39 in mussel byssus 1:310
see also chitosan/MMT in teeth 1:18
composites collagen composites
with alumina 2:142 see also collagen nanocomposites
with collagen 1:264 from chromium-containing
chitosan/MMT composites 2:36, wastes 1:266–270
43–61 from chromium-free
antimicrobial properties 2:45, wastes 1:263–266
58–59, 250 with silk 1:231, 235–236
applications 2:59–61, 250 collagen hydrogels 2:268
barrier properties 2:55–57 collagen nanocomposites 2:267–270,
mechanical properties 2:49–52, 50 274
preparation and colloidal force
characterization 2:43–49, 240, microscopy 1:183–201, 191–192
241, 242 combined composite materials
rheological properties 2:59 (defined) 1:124
in starch matrix 2:191–193 compact bone 1:13
thermal stability 2:53–55 composite matrices see matrices
water swelling properties 2:46, composite reinforcement see
57–58 reinforcement (fillers)
chondrocytes 2:261, 270, 278, 279, composite thickness 1:101
283 composites, definition and
chromium-containing proteinaceous classification 1:69–73, 121–124,
wastes composites 1:262–263, 162, 226
266–270 biocomposites 1:229, 261–262
clay aerogel 1:237–238 nanocomposites 2:3
clay-reinforced nanocomposites see compression molding 1:227–229
montmorillonite (MMT) conductive composites
composites with carbon nanotubes
coconut fibre see coir (CNT) 1:248, 2:211, 212, 213,
CoFe2O4 2:37 217–218, 219, 246–247
coir (coconut fibre) 1:68 silk-based 1:247–248
cellulose nanocrystals from 2:21 starch-based 2:195–197, 211, 212,
composition 1:43, 90, 126, 144 213, 217–218, 219
production and cost 1:39, 120 cone calorimetry 1:93, 94, 97, 98, 102,
properties 1:40, 41, 42, 44 105, 108
water sorption 1:46, 88, 142, construction industry 1:80, 81, 87
143–144, 174 contraction, spider silk 1:29, 289–294
Subject Index 335
controlled release see drug delivery dahllite 1:11, 15
systems date palm (Phoenix dactylifera) 1:42,
Corchorus capsularis see jute 43, 46, 54, 2:21
corn gluten meal (CGM) 1:165, 204, decussation, in dental tissue 1:16, 18
207, 208, 209–210 dendrimer immobilization 1:249
corn shell (husks) 1:55, 109, 109–110 density, of natural fibres 1:39–40,
corn starch 1:131, 172, 2:198 41, 88
see also waxy maize starch dental adhesives 1:322–323
cornstalk 1:40, 42, 43, 46, 55 dentine (dentin) 1:14, 17–18
cotton 1:67 dentition 1:16–17
cellulose nanocrystals from 2:21 Derjaguin–Muller–Toporov (DMT)
composition 1:43, 90, 144 theory 1:198
production and cost 1:39 desorption see water sorption
properties 1:40, 41, 42, 42–43, 88 dialectric properties 1:53, 2:218, 219
water sorption 1:46, 88, 143, 144, diammonium polyphosphate
150 (DAP) 1:108–109
cotton fibre composites 1:129, 164, 171 dichlorotriazines, as coupling
cotton stalks 1:40, 42, 43, 46, 55 agents 1:171
coupling agents 1:166–177 3,4-dihydroxyphenyl-L-alanine
bio-based 1:171–177 see DOPA
in byssus fibres 1:320 dispersion, filler 1:241, 245, 2:236,
dichlorotriazines 1:171 245–246, 247
isocyanates 1:169, 169–170, DOPA (3,4-dihydroxyphenyl-L-
171–172 alanine) 1:309, 311, 319–322, 323,
maleated 1:170–171, 2:96, 107–108 324
silanes 1:156–157, 167–169 double diffraction see birefringence
crack bridging 2:127, 128–129 drug delivery systems 1:246, 2:60–61,
crack resistance 2:120–127 262, 271, 276, 278
theoretical dyes, adsorption of organic 2:58, 60
measurement 2:127–131
cryocrushing (fibrillation Elaeis guineensis see oil palm
process) 2:10 elastic modulus (Young’s modulus,
crystallinity 1:44, 91 tensile modulus)
effect of plasticizers 2:75–78, 87 see also mechanical properties
spider silk 1:277, 286, 287, 289 cellulose 1:41, 2:5–7, 17, 24–25
crystallization behaviour 2:78–80, 83 determining 2:50
CTAB-modified MMT 2:46 modelling 2:118–120
curaua (Ananas erectifolius) 1:52–53 natural fibre composites, examples
cellulose nanocrystals from 2:21 of improvements in 1:163,
composition 1:43 164–165
production and cost 1:39 natural fibres 1:41, 42, 88, 128
properties 1:40, 41, 42, 46 effect of anisotropy 1:77, 79–80
curaua (nano)composites 1:174, effect of moisture 1:151–154
2:170 thermoplastic and thermoset
cyclic contraction, spider resins 1:123
silk 1:292–294 elastin 2:258, 262–263
336 Subject Index
elastin nanocomposites 2:272, fillers see reinforcement (fillers)
273–278 films
electrical conductivity see conductive cellulose 1:186, 2:16–17, 23–25
composites chitosan-based 2:44–46, 51–52,
electrical insulation 1:53 56–57, 250
electrochemical sensors 2:60 collagen/natural polymer
electrospun fibres 1:214, 246–247, hybrid 1:264–266
294–295, 2:104–106, 272–273, 274 soy protein-based 2:106–107
elephant grass (Pennisetum spider silk protein 1:295
purpureum) 1:41, 42, 51–52 starch-based 1:264–266, 2:189,
enamel, tooth 1:14–17, 18 190–191, 249
environmental impact superhydrophobic 2:164–166
of bio-based materials 1:260–261 thin films 1:186–188, 242, 2:16–17,
of vegetable fibres 1:127 23–25
enzyme immobilization 1:248–249, titanium dioxide (TiO2)
295–296, 2:35, 39 porous 2:161, 162–163
enzyme treatments 1:175–176, 2:13 zein 1:210–212
epoxy composites 1:54, 76, 112, 163, flagelliform silk 1:26, 28, 275, 285,
231, 234–235 289–290
epoxy resins 1:98 flame retardation 1:106–113, 2:23
equilibrium moisture content 1:141, matrices 1:106–110
143–144 natural fibres 1:110–112, 173
ethanolamine, as surface treatments 1:112–113
plasticizer 2:199–200 flammability 1:88–106
exfoliation 2:42–43, 70, 74, 84, 235 see also flame retardation; thermal
extracellular matrix (ECM) stability
proteins 1:323–324, 2:264–265 biodegradable matrices 1:94–96
composites 1:99–106
FA see furfuryl alcohol natural fibres 1:88–91
fatty acid derivatives, as coupling natural rubber 1:96–97
agents 1:173 performance
Fe3O4 nanoparticles 1:239, 2:37, 39 requirements 1:99–100
feldspar particles 2:217–219 thermoplastics 1:92–94, 93
fibre bundles 1:47–48 thermosets 1:97–98, 98
fibre length 1:73–74, 100, 101–102, 106 flax (Linum usitatissimum) 1:67
fibre orientation 1:64, 73–75, 77–80, 91 cellulose nanocrystals from 2:21
fibre saturation point 1:142–144 composition 1:43, 90, 126, 144
fibrin 2:258–259, 263 production and cost 1:39, 120
fibrin glue (sealant) 2:263, 278–279, 280 properties 1:40, 41, 42, 76, 90
fibrin nanocomposites 2:278–280 water sorption 1:46, 88, 142, 144,
fibrinogen 2:263 149, 150
fibroblast growth factors 2:272 flax fibre composites 1:92, 164, 165
fibroblasts 2:283 applications 1:129, 131
fibroin proteins 1:221, 223–226 effects of fibre orientation 1:79
fibrous composite materials with FA/PFA 2:171–172, 172
(defined) 1:124 flame retardation 1:107–108
Subject Index 337
flammability 1:90, 102, 102, glycerol 2:70, 70
104–106, 105, 2:172 see also plasticizers
poly(lactic acid) (PLA) 1:76, 173, in chitosan/MMT films 2:48, 52
184, 185, 199–200 in starch nanocomposites 2:203,
food packaging 1:211–212, 2:107, 208, 209
248–250 pretreatment of carbon
foot proteins see mussel foot proteins nanotubes 2:211, 212, 214
fracture process zone 2:127–128, in zein films 1:211
129–131 glycosaminoglycans (GAG) 1:236,
freeze drying 1:229, 2:16 264, 2:269–270
fungal modifications of fibres 1:174, gold nanoparticles 1:239, 248, 249,
176–177 2:37, 39
furan 2:150–152 graphite, expandable 1:107–108
furfural 2:108 grinding (fibrillation process) 2:9–10
furfuryl alcohol (FA) 1:174, growth bands, nacre 1:22, 2:114, 123,
2:153–154 131
see also poly(furfuryl alcohol) guar gum, as coupling agent 1:175
furfuryl palmitate 2:172–173
haemostatic agents 2:261, 278
gelatin 1:262, 2:257–258, 261–262 Hailwood–Horrobin
gelatin (nano)composites model 1:145–146
prepared by solution Haliotis rufescens see red abalone
intercalation 2:241–242 Harversian system 1:13
with silk 1:231, 236 heat of wetting 1:149
for tissue engineering 2:250–251, hemicellulose 1:41, 66, 89, 90,
270, 271–273, 274, 275, 125, 126
283–285 effect on cellulose
gelatin hydrogels 2:271, 272 defibrillation 2:11
Gelfoam 2:271 hemostatic agents 2:261, 278
gene delivery materials 1:325 hemp (Cannabis sativa) 1:68, 76, 162
glass fibre 1:123–124 cellulose nanocrystals from 2:21
production and cost 1:39, 120 composition 1:43, 90, 126, 144
properties 1:39, 41, 41, 42, 87, 88 odour 1:48
glass fibre composites production and cost 1:39, 120
hybrid composites 1:75, 235 properties 1:40, 41, 42, 76–77
mechanical properties 1:78–79, 79, thermal stability and
105 flammability 1:90
thermal stability and water sorption 1:46, 88, 144
flammability 1:97, 102–106, 105 hemp fibre composites 1:50, 92,
glass transition temperature 1:93, 164, 165
196–197, 2:81, 83, 85 applications 1:129
globulins 2:99 effects of fibre orientation 1:78, 79
gluten flame retardation 1:107
corn gluten meal (CGM) 1:165, henequen (Agave fourcroydes) 1:68,
204, 207, 208, 209–210 232
wheat gluten 1:162 Hibiscus cannabinus see kenaf
338 Subject Index
hierarchical structures hydroxyapatite (HA, HAp) 1:11, 15,
biomimetic duplication 1:29–31, 2:267
2:48, 141–145 hydroxyapatite (HA, HAp)
bone 1:8–14, 30 composites 2:250–251
lignocellulosic fibres 2:2–4 with chitosan 2:37, 250
mechanical advantages 1:13–14, with collagen 2:268, 269
18, 23, 28–29, 2:4–8, 115 with gelatin 2:250–251, 271
nacre 1:18–23, 2:114–115 with silk 1:232, 237, 242–244,
spider silk 1:24–29 296
teeth 1:14–18 with soy protein 2:99
high-density polyethylene 2-hydroxyethyl cellulose
composites 1:55 (HEC) 1:264–266, 267–270
histidines, in mussel byssus 1:310 N-(2-hydroxyethyl)formamide
hop (Humulus lupulus) 1:39, 40, 42, (HF) 2:197–199
43, 46, 54 hydroxypropyl alkaline lignin
humidity (HPL) 2:102
see also moisture content; water hygroscopicity 1:155–157,
sorption 2:170, 249
and interfacial see also water sorption
adhesion 1:196–197, 2:223–224
and mussel byssus fibres immiscible nanocomposites 2:42
1:314–315 immobilization
and nanocellulose 2:7, 17 applications 1:248–249, 295–296,
and spider silk 1:289–294 323, 2:35, 39
Humulus lupulus see hop immunosensors 1:323
Hunter–Schreger bands 1:16 implants, medical 2:260–261
hybrid composites in situ intercalative
artificial nacres 2:142–143 polymerization 2:41, 174–177,
natural fibre/glass fibre 1:75, 235 237–240
soy protein injection molding 1:227
nanocomposites 2:93–100 inorganic fillers 1:39, 238–239,
hybrid films 1:264–266 242–245, 2:215–220
hybrid polymer matrices 2:51, 55, see also metal nanoparticles; metal
57–58, 60 oxide nanoparticles;
hydrochloric acid, in cellulose montmorillonite (MMT) clay
hydrolysis 2:22 inorganic matrices 1:232, 237–238
hydrogels insulating properties 1:40, 53, 55
chitosan 2:39, 60–61 intercalation 2:41–43, 94, 235,
collagen 2:261, 268 237–246
fibrin 2:278–280 melt intercalation 2:41–42, 94,
gelatin 2:262, 271, 272 237, 243–246
microfibrillated cellulose 2:15 in situ polymerization 2:41,
mussel foot protein-based 1:323 174–177, 237–240
spider silk 1:295 from solution 2:41, 47–48, 237,
hydroxyaluminium cations, 240–243
intercalated 2:47–48 thermodynamics 2:243, 244
Subject Index 339
interfacial interactions 1:73, 163–166, kapok (Ceiba pentranda) 1:40, 42
183–201 kenaf (Hibiscus cannabinus) 1:40, 41,
cellulose nanocrystals 2:22 42, 43, 46, 90
colloidal force kenaf fibre composites 1:92, 164, 173
measurement 1:186–189,191–197 applications 1:129, 131
effect of FA/PFA 2:166, 171–172, kinetics of water sorption 1:149–151
173, 174
in fibre bundles 1:47–48 laccases 1:175–176
interfacial shear stress lactic acid 2:46, 55, 58, 60
(IFSS) 1:184, 197 see also poly(lactic acid)
model surfaces 1:190–191 lady’s finger see okra (Abelmoschus
in nacre 2:118–119 esculentus)
Nardin–Schultz model 1:184, laminated composite materials
199–200 (defined) 1:124
in silk composites 1:235, 237, laponite RD 2:192–193, 241, 249
242–244, 247 layered double hydroxides 2:251
in starch layered silicates see montmorillonite
nanocomposites 2:214–215, (MMT) clay
215–217, 223–224 leaf fibres 1:5, 65
work of adhesion 1:197–198 leather industry wastes 1:258–260,
interfacial shear stress (IFSS) 1:184, 197 262–263
intumescents 1:111–112 ligament repair 1:236, 246–247
ionic liquid-plasticized starch 2:195–197 lignin 1:41, 66, 2:178–179
isocyanate coupling agents 1:169, FA-modified 2:170–171
169–170, 171–172 natural fibres composition 1:90,
isora (Helicteres isora) 1:40, 41, 42, 125, 126
43, 46, 48–50 thermal stability and
flammability 1:89–90
Johnson–Kendall–Roberts (JKR) water sorption 1:142, 148
theory 1:198 lignin, hydroxypropyl alkaline
jute (Corchorus capsularis) 1:67 (HPL) 2:102
composition 1:43, 90, 126, 144 limiting oxygen index (LOI) 1:91
FA-modified 2:171 Linum usitatissimum see flax
production and cost 1:39, 120 liquid crystalline silk 1:27–28
properties 1:40, 41, 42, 76 low-density polyethylene (LDPE)
thermal stability and composites 1:171–172, 2:194–195
flammability 1:90–91 Luffa cylindrica 1:41, 43, 53–54, 2:21
water sorption 1:46, 88, 127–128, lumen 1:40
144, 149, 154 lysine diisocyanate (LDIC) 1:172
jute fibre composites 1:79, 92,
127–128, 164, 232–233 magnesium hydroxide 1:106–107
applications 1:131 magnetic nanoparticles 1:239, 2:37, 39
with coupling agents 1:171, 175 maize see corn; waxy maize starch
thermal stability and maleated polymers 1:170–171, 2:96,
flammability 1:90, 97, 107–108
102–106, 105 marine applications 1:99–100
340 Subject Index
matrices 1:64, 69–71, 122, 123 metal nanoparticles
flammability 1:92–98, 106–110 in chitosan-based composites 2:37,
inorganic 1:232, 237–238 39, 49, 59
Matriderm 2:276, 277 in silk-based composites 1:239,
Maugis parameter 1:198 248, 249
mechanical properties in starch-based composites 2:219
see also mechanical properties of metal oxide nanoparticles 1:242, 2:37,
(nano)composites 39, 159, 219–220
cellulose 2:4–8, 7, 16–17, 23–25 MFC see microfibrillated cellulose
in hierarchical structures 1:13–14, micro wincyette fibre 1:71
18, 23, 28–29, 2:4–8, 115 microbonding test 1:189
mussel byssus fibres 1:308, microfibril angle 1:41–43, 76, 76, 78,
313–315 125, 142
nacre 1:23, 2:115–120 microfibrillated cellulose
natural fibres 1:44–45, 46, 47–48, (MFC) 2:8–17, 101–102
76, 76–77, 79, 87, 88, 127–128, 128 aqueous suspension 2:14–16
chemically-modified 2:170–171 films 2:16–17
effect of water morphology 2:11–12, 12
sorption 1:127–128, 151–155 preparation 2:8–10
specific mechanical pretreatments 2:12, 12–14
properties 1:40–43, 41, 42 microfibrillated cellulose (MFC)
silks 1:28, 28–29, 279–288, 280 nanocomposites 2:101–102,
mechanical properties of 202–203
(nano)composites 1:75–77, 105, 2:50 microfibrils 1:41–43, 66, 2:3–4
chitosan/MMT 2:49–52, 50 see also microfibrillated cellulose
collagen-based 1:265–266, mineral, bone 1:11
268–270 mineral fibres 1:66, 125
effect of anisotropy 1:77–80 minor ampullate (MI) spidroin
PFA-based or - silk 1:26, 28, 275
modified 2:170–171 mirror-like films 1:245
soy protein nanocomposites 2:98, Miscanthus (China reed) 1:40, 41, 42,
101–102, 220–221 43, 46, 50–51
starch/carbon nanotubes MMT see montmorillonite (MMT) clay
(CNT) 2:212, 214–215, 216 modelling
starch/nanocellulose 2:203–204, deformation of nacre 2:117–120,
205, 207 131–136
starch/nanoclay 2:192–193, 194, fracture of nacre 2:127–137
194, 198–199, 200–202 large microstructures in
starch-reinforced nacre 2:138–141
nanocomposites 2:220–225, tablet junction failure in
226, 228, 229 nacre 2:134–137
starch/starch 2:208, 209 thermodynamics of melt
medical adhesives 1:322–323, intercalation 2:244
2:278–279, 280 toughness 2:127–131
melt intercalation 2:41–42, 94, 237, water sorption of natural
243–246 fibres 1:144–146
Subject Index 341
moisture content modelling deformation and failure
see also humidity; water sorption; patterns 2:131–141
water swelling properties modelling toughness 2:127–131
mussel byssus fibres 1:314–315 structure 1:18–23, 2:114–115
starch/nanoclay nacre-like composites 1:29–30, 2:48,
composites 2:81–84, 86–87, 196 51–52, 141–145
moisture regain by natural fibres 1:46 nanocellulose see cellulose
montmorillonite (MMT) clay 2:36, nanocrystals; cellulose
38–41, 41, 72 nanoparticles; microfibrillated
montmorillonite (MMT) cellulose
composites 2:41–43, 235, 237–246 nanoclay see montmorillonite (MMT)
see also intercalation clay; montmorillonite (MMT)
applications 2:59–61, 249, 250, 251 composites
with chitosan (see chitosan/MMT nanocomposites, definition and
composites) overview 2:3, 234–236
with gelatin 2:241–242 nanocrystalline cellulose see cellulose
with PFA 2:173–175 nanocrystals
with silk 1:232, 245 nanofibrillar cellulose see
with soy protein 2:94–96, 244 microfibrillated cellulose (MFC)
with starch 2:191–202, 219, 249 nanofillers, definition and types 2:3,
Mooney–Rivlin model 1:315 37, 38, 39
mother of pearl see nacre nanoparticles
Musa textilis see abaca see also cellulose nanocrystals;
mussel byssus fibres 1:305–320 cellulose nanoparticles; metal
adhesive properties 1:311, nanoparticles; metal oxide
319–321, 321–323 nanoparticles
applications 1:321–325 inorganic 1:238–239
mechanical properties 1:308, starch 2:205–210, 220–229
313–315 Nardin–Schultz model 1:199–200
source 1:306–308 natural fibre composites
structure and see also individual plant fibres
composition 1:308–313, 309, anisotropy 1:73–75, 77–80
317–319 applications 1:80–81, 129–137
thermal properties 1:315–317 flame retardation 1:112–113
mussel foot proteins (fp) 1:309, flammability 1:99–106
319–321, 320, 322 with PFA 2:169–173
mussel adhesive proteins, reported combinations 1:92
(MAP) 1:323–325 natural fibres (animal) 1:6, 66, 125
Mytilus species 1:307–308, 309, 311, see also mussel byssus fibres; silk;
314, 318, 319–321, 323 spider silk; wool
natural fibres (mineral) 1:66, 125
nacre 1:18–23, 2:113–146 natural fibres (vegetable) 1:4–6, 5,
artificial 1:29–30, 2:48, 51–52, 65–68, 124–129
141–145 classification 1:125
deformation 2:115–120 as composite reinforcement
fracture 2:120–127 materials 1:37–57
342 Subject Index
natural fibres (vegetable) (continued) orientation, reinforcement/filler
advantages 1:38–44, 64, 72, 87, carbon nanotubes 2:247
126–129 fibres 1:64, 73–75, 77–80, 91
disadvantages 1:44–48, 64, 72, osteons 1:13
87–88, 126–129, 2:1–2 oxidation pretreatment 2:14
composition 1:43, 90, 125, 126, 144 oxygen permeability 1:212, 2:57, 107,
density 1:39–40, 41, 88 228
FA-modified 2:169–173
flame retardation 1:110–112, 173 packaging materials 1:211–212,
flammability 1:88–91 2:59, 106–107, 248–250
mechanical properties 1:44–45, palm fibres
47–48, 76, 76–77, 79, 87, 88, date (Phoenix dactylifera) 1:41,
127–128, 128 42, 43, 46, 54, 2:21
effect of chemical oil (Elaeis guineensis) 1:75, 163
modification 2:170–171 vakka (royal palm, Roystonea
specific 1:40–43, 41, 42, 128 regia) 1:41, 42, 46, 53
effect of water palmitic acid 1:211
sorption 1:127–128, palmitic acid, furfuryl ester
151–155 of 2:172–173
physical properties 1:87, 88, 128 Panicum virgatum see switchgrass
production and cost 1:39, 120, parallel exponential
126–127 kinetics 1:149–150
sources 1:48–56, 66–68 particle composite materials
structural anisotropy 1:79–80 (defined) 1:124
structure 1:74–75, 2:2–5 particleboards 1:130
thermal stability 1:46, 88–91, 129, pea hull 2:21
173 pea starch 2:102, 203–204, 206,
variability of 1:44–45, 73, 128 207, 215–217, 218, 219–222,
water sorption (see water sorption) 224–227, 228
natural polymers, definition and pearl oyster (Pinctada
examples 1:2–3, 2–4, 4, 164, 165, margaritifera) 2:120,
260–261 122–123, 125
natural rubber 1:96–97 pectinases 1:175
natural rubber composites 1:48, 75, pectin(s) 1:187–188
236–237, 2:227–229 effect on cellulose
nerve regeneration 2:270 defibrillation 2:11
nettle (Urtica dioica) 1:40, 41, 42, 43, effect on mechanical
46 properties 2:17
nitrile rubber composites 1:230 natural fibres composition 1:90
nonprismatic enamel see prismless and odour 1:48
enamel pen shell (Pinna nobilis) 2:120, 123
Pennisetum purpureum see elephant
odour of natural fibres 1:48 grass
oil palm (Elaeis guineensis) 1:75, 163 permeability of composites see barrier
okra (Abelmoschus esculentus) 1:43, properties
46, 52 PFA see poly(furfuryl alcohol)
Subject Index 343
phenol/formaldehyde (PF) with gelatin 2:272–273, 274, 275,
composites 1:163 284–285
phenolic resins 1:98 with silk 1:231, 233–234
Phoenix dactylifera see date palm with starch/PLA 2:224–225, 226
Phytagel-modified soy protein poly(e-caprolactone-co-D,L-lactide)
(PH-SPC) 2:95–96 1:241
Pinctada margaritifera see pearl oyster polycarbonate (PC) 1:93, 163,
pineapple (Ananas comosus) 1:39, 231, 235
40, 41, 42, 43, 46 poly(dimethylsiloxane)
pineapple leaf fibre (PALF) (PDMS) 1:267–270
composites 1:92, 163, 164, 170, 171 polyester [poly(ethylene
Pinna nobilis see pen shell terephthalate)] 1:93
piriform silk 1:275 polyester amide (PEA) 1:164
plant fibres see natural fibres polyester composites 1:51–52, 53, 54,
(vegetable) 55, 164
plasma, blood 2:258–259, 263 with coupling agents 1:170, 171
plasticizers 1:210–211, 2:69–71, 72–89 polyethylene (PE) 1:93, 94
effect of molecular size 2:72–74 polyethylene (PE) composites
effect of molecular with coupling agents 1:170,
symmetry 2:74–75 171–172
effect on crystallization 2:75–80, with natural fibres 1:55, 163
87, 200, 201 with silk 1:236–237
effect on mechanical with starch/MMT 2:194–195
properties 2:200–202 poly(ethylene glycol) (PEG) 2:259,
effect on nanostructure 2:243–244 268, 271
morphology ‘‘threshold’’ 2:89 poly(ethylene oxide) (PEO) 2:105
novel plasticizers 2:195–196, poly(ethylene terephthalate)
197–199 (PET) 1:93
pollutant adsorption 2:60 see also polyester composites
polyamide-6 1:53, 93 poly(furfuryl alcohol)
poly(butyl acrylate) 2:44, 51, 55, (PFA) 2:153–180
57–58 see also furfuryl alcohol
poly(butylene adipate-co- carbon membrane
terephthalate) 1:164 synthesis 2:159–163
poly(butylene succinate) (PBS) 1:164, carbon nanotube modification/
172, 230–232 composites 2:163–169
poly[(butylene succinate-co-(butylene formation 2:153–154
adipate)] (PBSA) 1:231, 234 natural fibre modification/
poly(butylene terephthalate) 1:93 composites 2:169–173
poly(e-caprolactone), maleated 2:96, porous carbon materials
107–108 synthesis 2:154–159
poly(e-caprolactone) (PCL) in situ polymerization with
(nano)composites nanoparticle
with cellulose 1:164 reinforcement 2:173–177
chitosan blend with MMT 2:45 wood modification 2:177–180
with collagen 1:264, 2:270, 274 poly(glycolic acid) 2:280
344 Subject Index
poly(hydroxybutyrate) (PHB) 1:164, poly(methyl methacrylate) (PMMA)
2:244, 249–250 composites
poly(hydroxybutyrate valerate) artificial nacres 2:142–143, 145
(PHBV) 1:164 with leather wastes 1:263
polyimide/montmorillonite with silk 1:231, 235
nanocomposites 2:240 polypropylene (PP)
poly(lactic acid) (PLA) 1:135, in polymer blends 1:232, 237
2:249–250 thermal stability and
colloidal probes 1:188, 190–191, flammability 1:93, 94, 95–96, 97
193 polypropylene (PP) composites
flammability 1:94–96, 97 applications 1:131, 231, 232
poly(lactic acid) (PLA) with coupling agents 1:170–171, 173
composites 1:164 flame retardation 1:107, 108–109,
biomedical applications 2:251 109
with collagen 1:264 with natural fibres 1:50–51, 54,
with coupling agents 1:170, 172, 163, 171, 232–233
173 with silk 1:231, 232, 232–233
flammability 1:97, 102–104 thermal stability and
with flax 1:76, 173, 184, 185, flammability 1:97, 101–104, 102
199–200 polystyrene (PS) 1:93, 94, 163
interfacial adhesion 1:185 poly(styrene-b-ethylene/butylene-b-
with Miscanthus 1:51 styrene) (SEBS) 1:241
with nanoclays (MMT, etc.) 2:58, poly(tetrafluoroethylene)
193–194, 237–240, 241, 243, 244, (PTFE) 1:93
250 polyurethane
packaging materials 2:249–250 (nano)composites 1:131, 241,
with rice straw 1:55 2:222–223, 280
with silk 1:231, 233, 240–241 flame retardation 1:108–110, 109
with starch 2:193–194, 224–225, poly(vinyl alcohol) (PVA,
226 PVOH) 1:164
poly(lactide-co-glycolide) with silk 1:232, 237–238
(PLGA) 1:246–247, 2:270, 276 with starch
poly(L-lysine) 2:251 nanoparticles 2:225–227, 228
polymer blends in starch/PVA blend
with nanoclay 2:195, 240–241 matrix 2:240–241, 247, 249
silk fibre-reinforced 1:232, poly(vinyl chloride) 1:93, 94, 163,
236–237 172–173, 263
polymer reinforcement see poly(vinylpyrrolidone)
reinforcement (fillers) (PVP) 1:267–270
polymeric matrices see matrices poplar wood 1:130
polymers, definitions and examples porosity of fibre bundles 1:47
natural (biopolymers) 1:2–3, 4, porous carbons synthesis 2:154–159
164, 165, 260–261 porous composite materials 1:229,
synthetic 1:2, 163, 164, 260 245–246
poly(methyl methacrylate) see also scaffolds (tissue
(PMMA) 1:93 engineering)
Subject Index 345
potassium persulfate (KPS) 2:44–45, relative permeability 2:56
51 rhectophyllum (Rhectophyllum
potato starch 2:222–223, 249 camerunense) 1:40, 41, 42, 43, 54
preprism enamel 1:15 rice husks 1:55, 107
prismatic enamel 1:15–16 rice straw 1:39, 40, 43, 46, 55, 163
prismless enamel 1:15 royal palm (vakka, Roystonea
properties of composites 1:71–73 regia) 1:41, 42, 46, 53
see also barrier properties; rubber (natural) 1:96–97
dialectric properties; insulating rubber (natural) composites 1:48, 75,
properties; mechanical 236–237, 2:227–229
properties of (nano)composites; rubber (synthetic) composites 1:55,
water swelling properties 230, 263
protein fibres 1:6, 66, 125
see also mussel byssus fibres; silk; salt leaching 1:229
spider silk; wool scaffolds (tissue engineering)
protein-based polymers (animal) chitosan-based 2:58, 60–61
see also collagen; fibrin; gelatin collagen-based 2:267–270, 274
isolation of 2:257–259 design requirements 2:255–257
structure and properties 2:259–263 elastin-based 2:273–278
surface modification 2:263–267 fibrin-based 2:278–280
protein-based polymers (vegetable) gelatin-based 2:271–273, 274, 275
see soy protein; zein silk-based 1:235–236, 240–241,
pullulan 2:223–224 241–242, 243, 244–247
pulp, tooth 1:14 spider silk-based 1:295
surface modification 2:263–267
radial enamel 1:16 seashells 1:18–23
rail industry 1:99 see also nacre
ramie (Boehmeria nivea) 1:67 sericin 1:220–221, 223
cellulose nanocrystals from 2:21 sericulture see silkworms
composition 1:43, 90 shear lag model analysis 2:125
production and cost 1:39 shellac, as coupling agent
properties 1:40, 41, 42, 46, 88 1:173–174
ramie fibre composites 1:70, 92, 165 shipping industry 1:99–100
rectorite (REC) 2:96 silane coupling agents 1:156–157,
red abalone (Haliotis 167, 167–169
rufescens) 2:120, 123, 125 silica composites
reflective films 1:245 with PFA 2:154–159
regenerative medicine see tissue with silk 1:244
engineering with soy protein 2:100
reinforcement (fillers) with spider silk 1:296
nanofillers 2:3, 37, 38, 39 silicates see montmorillonite (MMT)
orientation clay; montmorillonite (MMT)
carbon nanotubes 2:247 composites; wollastonite (CaSiO3)
fibres 1:64, 73–75, 77–80, 91 silk 1:30, 219–249
types 1:72–73, 100–101, 122–124, see also spider silk
226 manufacture 1:222–226
346 Subject Index
silk (continued) sorghum 1:40
properties 1:66, 221–222, 280–281, sorption see water sorption
286 sorption isotherms 1:141–144
regenerated 1:223–226, 239–246, sound proofing 1:40, 55
243 soy flour (SF) 1:165, 2:91
sources 1:220 soy globulin 2:99
structure 1:220–221, 276–277, 277 soy protein 2:91–93
silk-based composites 1:164, 226–249 soy protein composites 1:70, 165
biosensors 1:248–249 soy protein nanocomposites 2:93–109
conductive 1:247–248 with alumina hydrate 2:99–100
silk as matrix 1:238–239, 241–246, applications 2:103–108
243 with calcium carbonate 2:97–99
silk as reinforcement 1:229–238, with carbon nanotubes 2:96–97
231–232, 239–241, 240 with clay 2:93–96, 244
silk-reinforced silk 1:246–247 with polymeric emulsion
silkworms 1:220, 222–223 particles 2:102
silver halide nanoparticles 1:238–239 with polysaccharides 2:100–102
silver nanoparticles with silica 2:100
with chitosan 2:37, 39, 49, 59 with starch 2:220–222
with silk matrix 1:239 soy protein nanofibres 2:104–106
with starch/MMT 2:219 soybean straw 1:40, 42, 43, 46, 55
silver nanoplates 1:245 specific mechanical properties of
sisal (Agave sisalana) 1:68 natural fibres 1:40–43, 41, 42, 128
cellulose nanoparticles from 2:16, spider silk 1:30, 220, 275–297
21, 24, 24 applications 1:294–296
composition 1:43, 90, 126 formation in vivo 1:278–279, 286,
production and cost 1:39, 120 288
properties 1:40, 41, 42, 46, 76, 88 mechanical properties 1:28, 28–29,
thermal stability and 279–288, 280
flammability 1:90–91 metal infiltration 1:283
sisal fibre composites 1:76, 92, 163, structure 1:24–29, 276–277,
164, 165, 231 283–289
applications 1:129, 131 supercontraction 1:29, 289–294
with coupling agents 1:170, 171–172 spider silk fibre composites 1:239,
PFA-modified 2:170–171 244, 296
thermal stability and sponge-gourd (Luffa
flammability 1:90, 97, 102–106, cylindrica) 1:41, 43, 53–54, 2:21
105 starch 1:3, 69, 2:186–191, 190
skin substitutes 1:264, 294, 2:268, starch composites 1:69–70, 71, 165
276, 277 see also starch nanocomposites;
skin wound healing 1:244 starch/nanoclay composites
soils, fibre-reinforced 1:78 with coupling agents 1:170, 172
solution intercalation 2:41, 47–48, flame retardation 1:108–109, 110
237, 240–243 starch nanocomposites 2:191–231
sorbitol 2:71, 71, 72 see also starch/nanoclay
see also plasticizers composites
Subject Index 347
applications 2:230, 249 straw (rice, soybean, wheat) 1:39, 40,
biodegradability 2:229 43, 46, 55, 163, 2:21
starch as matrix 2:191–220 straw (wheat) composites 1:92, 131,
with carbon 164
nanotubes 2:210–215, 247 sugar beet pulp 2:21
with cellulose sugar cane bagasse 1:174, 2:21, 170
nanoparticles 2:202–205 sulfuric acid, in cellulose
with inorganic hydrolysis 2:22–23
nanofillers 2:215–220 sunflower stalks 1:130
with starch supercontraction, spider silk 1:29,
nanoparticles 2:205–210 289–294
starch as reinforcement 2:220–229 superhydrophobic films 2:164–166
in natural rubber 2:227–229 surface fibres 1:65
in poly(lactic acid) 2:224–225 surface modification
in poly(vinyl montmorillonite (MMT)
alcohol) 2:225–227 clay 2:40–41
in pullulan 2:223–224 protein-based polymers
in soy protein isolate 2:102, 2:263–267
220–222 surgical adhesives 1:322, 2:278–279,
in starch matrix 2:205–210 280
in waterborne swelling properties 2:46, 57–58,
polyurethane 2:222–223 227–228
starch-based packaging switchgrass (Panicum
materials 2:249 virgatum) 1:39, 40, 41, 42, 43,
starch/collagen hybrid 46, 50
films 1:264–266 synthetic polymers, definition and
starch/nanoclay examples 1:2, 163, 164, 260
composites 2:191–202
applications 2:249 tablet structure in nacre 2:114–115,
effect of plasticizers 2:69–70, 116–117
72–89 brittle fracture 2:134
chemical interactions 2:75–80 junction failure
moisture content 2:81–84, modelling 2:134–137
86–87, 196 waviness 2:116, 139–141, 145
physical interactions 2:70–75 tannic acid, adsorption of 2:58, 60
synergistic interactions 2:84–89 teeth 1:14–18, 30
intercalation processes 2:240–241, temperature-dependence of properties
243–244 see thermal stability
mechanical properties 2:192–193, template-inhibition strategy 1:29–30
194, 194, 198–199, 200–202 TEMPO-mediated oxidation 2:14
stem cells 2:271–272, 281, 283 tensile modulus see elastic modulus
stem fibres 1:65 tensile properties see mechanical
stiffness properties
see also mechanical properties 2,2,6,6-tetramethylpiperidin-1-oxyl
composites 2:50, 118 radical see TEMPO-mediated
natural fibres 1:76, 77, 79, 79 oxidation
348 Subject Index
textile structure gelatin-based 2:271–273,
reinforcements 1:100, 102–104 274, 275
thermal insulation 1:40 silk-based 1:235–236, 240–242,
thermal stability 243, 244–247
see also flame retardation; spider silk-based 1:295
flammability surface modification 2:263–267
cellulose 1:89, 2:14, 23 titanium, infiltration into spider
chitosan/MMT silk 1:283
composites 2:53–55 titanium dioxide (TiO2)
mussel byssus fibres 1:315–317 nanocomposites 1:242, 2:37, 159
natural fibres 1:46, 88–91, 129, 173 porous film 2:161, 162–163
PFA composites 2:175, 178 top shell (Trochus niloticus) 2:120,
starch 2:189–190 123, 125
thermoplastic composites 1:70–71, tougheners 1:234–235
100, 165 toughness
flame retardation 1:106–110, modelling 2:127–131
112–113 of nacre 2:117, 120–127
flammability 1:101–104 of silks 1:279–280, 282, 283, 284
thermoplastic matrices 1:64, 106–107, transparent films 1:245, 2:17
122, 123 transport industry 1:99–100
flammability 1:92–94, 93 see also automotive industry
thermoset composites 1:100, TRIAB-modified MMT 2:51, 55,
104–106, 112–113 57–58
thermoset matrices 1:97–98, 98, 106, tricetadecylmethylammonium
122, 123 bromide see TRIAB
thickness, composite 1:101 Trochus niloticus see top shell
thin films tunicin 2:6–7, 21, 22
cellulose 1:186–188, 2:16–17,
23–25 ultimate stress
hydroxyapatite/silk fibroin 1:242 mussel byssus fibres 1:314, 315
polysaccharide, preparation plant fibres and composites 1:76,
of 1:186–188 76, 79, 79
thixotropy, in MFC suspensions ultrasonication 2:10, 15, 95
2:15 unsaturated polyester
thrombogenicity 2:260, 265 composites 1:49–50, 54, 164, 235
tissue engineering 2:255–257, flammability 1:104–106, 107
280–285 unsaturated polyester resins 1:97–98
cell adhesion 2:58, 265–266, 267, urea/formaldehyde (UF)
272–273, 280–281, 283, 284 adhesives 1:130
cell adhesion agents 1:323–325 Urtica dioica see nettle
scaffolds
chitosan-based 2:58, 60–61 vakka (royal palm, Roystonea
collagen-based 2:267–270, 274 regia) 1:41, 42, 46, 53
design requirements 2:255–257 variability of natural fibres 1:44–45,
elastin-based 2:273–278 73, 128
fibrin-based 2:278–280 vascular grafts 2:272, 276
Subject Index 349
vegetable fibres see natural fibres wollastonite (CaSiO3) 1:243, 244–245
(vegetable) wood (wood fibre) 1:65, 76, 2:2–4, 21
velvet leaf (Abutilon acetylation 1:155–156
theophrasti) 1:40, 42, 43, 46, 54–55 fibre saturation point 1:143
vermiculite 2:238–240 furfurylation 2:177–180
vertical flame spread 1:91 wood adhesives 2:103–104
vinyl ester resin composites 1:175 wood fibre composites 1:90, 92, 164,
vinyl ester resins 1:97–98 165
applications 1:129, 131
water barrier properties 1:211, 212, effects of fibre orientation 1:79
2:223, 224 flammability 1:102, 107, 109,
see also water vapour permeability 109–110
water sorption with maleated coupling
see also humidity; moisture agent 1:170
content; water swelling stiffness 1:77
properties wood flour composites 1:92, 172–173
of (nano)composites 1:127–128, wool 1:66, 91, 102–104
235, 2:195 wool fibre composites 1:92, 97
soy protein-based 2:92, work of adhesion 1:184, 197–198, 198
100–101, 102, 103 wound healing 1:244, 2:268, 279
of natural fibres 1:45–46, 46, 88, woven fabric reinforcements 1:100,
140–158 102–104
cell wall swelling 1:148
effect of PFA xylitol 2:70–71, 71, 72
modification 2:171 see also plasticizers
effect on mechanical xyloglucans 1:186–187, 192
behaviour 1:127–128,
151–155 Young’s modulus see elastic modulus
hysteresis 1:146–148
kinetics 1:149–151 zein 1:204–214
models 1:144–146 applications 1:213–214
reduction of 1:155–157, 174 as coupling agent 1:173
water swelling properties 2:46, 57–58, films 1:210–212
227–228 manufacture 1:207–210
water vapour permeability 1:212, structure 1:205–207
2:56–57, 107, 208, 228, 249, 250 modifications 1:212–213
see also water barrier properties zein/soy protein
waterborne polyurethane nanocomposites 2:105–106
(WPU) 1:241, 2:222–223 zeolite 2:159
waxy maize starch 2:206–210, zinc, infiltration into spider silk
223–224, 229 1:283
wetting, heat of 1:149 zinc oxide (ZnO) nanoparticles 2:37,
wheat gluten 1:162 219–220
wheat starch composites 1:170 zirconium glycine-N,N-
wheat straw 1:39, 43, 46, 2:21 dimethylphosphonate
wheat straw composites 1:92, 131, 164 (ZGDMP) 2:215–217, 218

Natural Polymers: Volume 1: Composites

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Natural Polymers

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r The Royal Society of Chemistry 2012

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Published by The Royal Society of Chemistry,

Registered Charity Number 207890

For further information see our web site at www.rsc.org

Natural polymers oﬀer an alternative solution to the growing environmental

RSC Green Chemistry No. 16

Maya Jacob John and Sabu Thomas

Chapter 2 Biomimetics: Inspiration from the Structural Organization

2.1 Introduction: Hierarchy and Structural Order 8

RSC Green Chemistry No. 16

Chapter 4 Relation between Structural Anisotropy in Natural Fibres

Chapter 5 Flame Retardant Characteristics of Natural Fibre

Chapter 6 Natural Fibre Composites: Automotive Applications 118

6.1 Introduction 118

Chapter 7 Water Vapour Sorption of Natural Fibres 140

7.1 Introduction 140

Chapter 8 Environmentally Friendly Coupling Agents for Natural

8.1 Introduction 161

Chapter 9 Probing Interfacial Interactions in Natural Fibre

9.1 Introduction 183

Chapter 10 Zein: Structure, Production, Film Properties and

10.1 Introduction 204

Chapter 11 Silk Fibre Composites 219

11.1 Introduction to Silk Fibres and their

12.1 Introduction 257

Chapter 13 Spider Silk: The Toughest Natural Polymer 275

13.1 Introduction to Natural Spider Silk Fibres 275

14.1 Introduction 305

Subject Index 330

Chapter 1 Nanocellulose: Potential Reinforcement in Composites 1

Chapter 2 Chitosan-based Nanocomposites 33

Chapter 3 Characterization of Molecular Interactions in Amylose

Chapter 4 Soy Protein Nanocomposites: Emerging Trends and

Chapter 5 Nacre from Mollusk Shells: Inspiration for

5.1 Overview and Microstructure of Nacre 113

Chapter 6 Nanocomposites from Furanic Derivatives 150

6.1 Furan and Furan Derivatives 150

Chapter 7 Starch Nanocomposites 185

7.1 Introduction 185

Chapter 8 Processing and Industrial Applications of Natural Polymer

8.1 Introduction 234

9.1 Introduction 255

Subject Index 294

Natural Polymers: An Overview

RSC Green Chemistry No. 16

Figure 1.1 Chart of world oil reserves.

Table 1.1 List of selected synthetic polymers.

1.2 Natural Polymer Research

1.2.1 Natural Fibres

Table 1.3 List of important natural ﬁbres.

1.2.2 Protein Fibres

Biomimetics: Inspiration from

Department of Civil Engineering, North Dakota State University, Fargo,

2.1 Introduction: Hierarchy and Structural Order