Holland 2018

PROGRESS REPORT
Silk www.advhealthmat.de
The Biomedical Use of Silk: Past, Present, Future

Chris Holland,* Keiji Numata,* Jelena Rnjak-Kovacina,* and F. Philipp Seib*
spend some time considering the motiva-

Humans have long appreciated silk for its lustrous appeal and remarkable tion of the research and the history of the
physical properties, yet as the mysteries of silk are unraveled, it becomes material that has led us to today.
clear that this outstanding biopolymer is more than a high-tech fiber. This The issue of motivation for studies
in the field of silk research is generally
progress report provides a critical but detailed insight into the biomedical use
divided into bottom-up, curiosity-driven
of silk. This journey begins with a historical perspective of silk and its uses, fundamental research and top-down, chal-
including the long-standing desire to reverse engineer silk. Selected silk struc- lenge-based activities.
ture–function relationships are then examined to appreciate past and current Fundamental silk research hinges on
silk challenges. From this, biocompatibility and biodegradation are reviewed the question, “what can we learn from
nature?” This is clearly a wider topic than
with a specific focus of silk performance in humans. The current clinical
silk itself, but the overall approach helps
uses of silk (e.g., sutures, surgical meshes, and fabrics) are discussed, as frame scholarly activities in the area.
well as clinical trials (e.g., wound healing, tissue engineering) and emerging We certainly have more to learn beyond
biomedical applications of silk across selected formats, such as silk solution, understanding the silk fiber itself, and
films, scaffolds, electrospun materials, hydrogels, and particles. The journey dozens of cross-disciplinary researchers
finishes with a look at the roadmap of next-generation recombinant silks, worldwide are using both simulation and
experimentation[1] to make concerted
especially the development pipeline of this new industry for clinical use.
efforts to understand the evolution,[2] pro-
cessing,[3] and performance of silk,[4] from
the molecule[5] to the material.[6] However,
1. Prologue as we broaden our interpretive horizons, we must remember
that silks are biological materials, and thus are defined by their
Before we begin to define the current state of the art sur- biology, before we attempt to transfer this knowledge to bioma-
rounding the field of silk-based biomaterials for (bio)medical terials, which are defined through their application.
use and look toward their future, we feel it is important to
Dr. C. Holland 2. Introduction

Department of Materials Science and Engineering
The University of Sheffield For the purpose of this progress review, we use the term silk
Sir Robert Hadfield Building, Mappin Street, Sheffield, South Yorkshire to refer to protein-based fiber-forming materials spun by living
S1 3JD, UK
E-mail: [email protected]
organisms. We also include in our terminology silk-inspired
Dr. K. Numata
proteins produced by recombinant approaches.
Biomacromolecules Research Team When studying silks, one must always appreciate that the
RIKEN Center for Sustainable Resource Science results derived from testing any naturally obtained biological
2-1 Hirosawa, Wako, Saitama 351-0198, Japan material are a product of both nature (its evolution) and nur-
E-mail: [email protected] ture (its environment), with the latter typically constraining
Dr. J. Rnjak-Kovacina the property space of the former (although exceptions
Graduate School of Biomedical Engineering
The University of New South Wales exist[4a,7]).
Sydney, NSW 2052, Australia The biological definition of a silk is a structural protein that
E-mail: [email protected] is spun into a fiber for use outside the body.[4a,8] In the wild,
Dr. F. P. Seib silks have undergone over 400 million years of “research and
Leibniz Institute of Polymer Research Dresden development” via natural selection, and after solutions to bio-
Max Bergmann Center of Biomaterials Dresden
Dresden 01069, Germany
logical challenges that range from predation (spider webs) to
E-mail: [email protected] housing (honey bees and wasps) and protection (silkworm
Dr. F. P. Seib cocoons).[4a,8] The ubiquity and widespread use of silk is a clear
Strathclyde Institute of Pharmacy and Biomedical Sciences testament to its success, especially as it has arisen numerous
University of Strathclyde times in independent convergent evolutionary events.[2c] Hence,
Glasgow G4 0RE, UK looking at how silk materials have evolved can not only deter-
The ORCID identification number(s) for the author(s) of this article mine their performance in the present, but can also reveal
can be found under https://doi.org/10.1002/adhm.201800465.
common design criteria and molecular “blueprints” for high
DOI: 10.1002/adhm.201800465 performance biological materials.[4a,9]
Adv. Healthcare Mater. 2018, 1800465 1800465 (1 of 26) © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.advancedsciencenews.com www.advhealthmat.de
Figure 1. Timeline: Milestones in the emergence of silk for biomedical applications.
In unraveling the properties of silk, we have also begun to by Shakespeare’s character, Nick Bottom, in A Midsummer
address common misconceptions regarding biological materials Night’s Dream, who says, “I shall desire you of more acquaint-
and their potential for industrial application. These are often ance, good Master Cobweb. If I cut my finger, I shall make bold
tarred with a brush of sample variability, suggesting that they of you.”[15]
are unsuitable for engineering or medical applications where However, the above examples describe the use of silk in its
consistency is key. However, recent studies now show that the unprocessed, natural state. A step forward in the utilitarian
variation previously observed is typically a manifestation of a evolution of silk came about with the realization that silk
silk’s exquisite responsiveness to its surroundings (making could be readily reprocessed into different forms. This was
silks incredibly “smart” materials).[4d,7a,10] Yet for uninitiated first performed at the macroscale by unwinding fibers from
researchers, this can sometimes become unwanted variation if the nonwoven composite cocoons of the silkworm Bombyx
the they fail to ensure consistent sample preparation or testing mori to create textiles. This skill originated in China, and
environments. Hence, biological diversity and plasticity offer direct archeological evidence confirms human interactions
several important lessons for those wishing to make the best with silkworm silk originating from the Neolithic period of
use of silk for their own applications. the 4th millennium BC, with the discovery of examples of cut
At the other end of the spectrum is the widely held belief cocoons and rudimentary looms at numerous archeological
that biological materials automatically qualify as “biocompat- sites.[16] Further archeological evidence suggests that the Indus
ible” materials. While these materials, including many silks, Valley civilization (in what is now Northern Pakistan) was also
are often biocompatible, simply labeling silk as “biocompatible” developing silk materials based on Antheraea silk. Therefore,
without context specific biocompatibility testing and critical sericulture—the act of rearing silkworms specifically for their
assessment of the available evidence is not in the best interest silk—can be estimated to have spread across South Asia from
of the field or, ultimately, patients. This mindset also perme- 5000–2000 BC.[17]
ates into the assumption that all natural materials are “green” Textiles produced from silk were truly a disruptive product,
which without appropriate and carefully considered environ- as they required both a unique material and highly sophisti-
mental analysis, the use of the phrase ultimately detracts from cated processing (programmable looms for weaving that were,
any potential impact of these materials. in essence, the progenitor of modern computing).[18] As such,
Once past our prejudices, at the interface of fundamental silk textiles were sufficiently valuable to become a formal cur-
and challenge-based activities sits biomimetics. This specifi- rency for Chinese soldiers at the edges of the empire and
cally looks to nature to reveal concepts, processes, and systems were used to barter with the locals for goods.[16] Nevertheless,
that can be applied to solve human challenges.[11] While the silk production remained a closely guarded secret within the
term “biomimetics” was only coined by American biophysicist Chinese empire for several thousand years, and when asked,
Otto Schmitt in the latter half of the 20th century,[11] humans traders would say it was “derived from the wool of sheep
have been looking to translate silk’s natural utility for their sprinkled with water and exposed to sunshine.” However, this
own use for millennia.[12] The simplest, most primitive forms product monopoly could not go unchallenged for long, and the
of mimicry are examples of imitation of the spider’s use of establishment of trade routes (the “silk roads”), and the apoc-
silk to catch prey, as seen in the Australian Aborigines’ use of ryphal industrial espionage that ensued, made silk technology
spider silk as fishing lines and New Guinean natives’ develop- available throughout the world. As a result, Bombyx mori silk
ment of fishing nets and bags.[13] However, the biological diver- has developed hand in hand with humans, through domestica-
sity of silk soon inspired humans to adapt silk for their own tion and artificial selection of the moths for over 4000 years.[16]
needs (e.g., ref. [14]), extending the silk phenotype beyond its This extensive history is a testament to the success and suit-
natural remit. Some of the first examples were the use of silks ability of this animal for large-scale industrial agricultural
medicinally by ancient Greeks and Romans, who bundled up development, as ≈980 billion animals are raised each year to
spider silk to treat wounds (Figure 1).[13] This was even noted produce ≈400 megatonnes of commercial silk.[19]
Across millennia, silkworm has been a luxury item for the elite. acid.[27] In subsequent years, a notable race began between
However, Claudius Galenus of Pergamon (c. 131 to c. 211 AD) Japanese and German researchers in the 1920s, with patents
was the first to document a potential medical application of the granted in 1924–1927[26a,28] and 1928,[29] respectively, for the
silk thread. Galenus gained a reputation for treating gladiators successful dissolution (and respinning) of artificial fibers using
whose tendons were severed in hand-to-hand combat and noted ZnCl2, Mg(NO3)2, and orthophosphoric acid as the main chao-
in his book De Methodo Medendi (150 AD) the use of several tropic agents. However, not until the 1930s did today’s familiar
materials as sutures, including linen. He writes, “in many places degumming using Na2CO3[30] and dissolution in LiBr appear.[26b]
under Roman rule you can obtain silk, especially in large cities The latter report clearly noted the potential of silk regeneration/
where there are many wealthy women. If there is no such oppor- reconstitution: “These solutions containing, if at all, only a small
tunity, choose from the material where you were living the least amount of salt, may be used in the known manner to produce
putrescible such as thin catgut.” Galenus’s teaching persisted artificial articles, such as fibers, films, or plastic masses.”
for centuries after his death but was eventually lost.[20] The war Beyond the replication of silk fibers for textile use, these feed-
surgeon Ambroise Paré (1510–1590) avoided cauterizing open stocks were originally intended for reprocessing into solid form
wounds with boiling oil and reverted to using vascular ligatures to harness silk’s excellent insulating properties[26b] and enable
made of silk or fine linen strips. However, only in 1869 did Joseph the casting of films (to make fabrics water and air imperme-
Lister introduce the first sterile silk suture into clinical practice.[20] able[31]). This was mainly because naturally derived materials
Throughout history, several alternative sources for textile were still superior in many aspects when compared to those
silk beyond the domesticated silkworm have been sought, from arising from the burgeoning field of industrial polymers.[32]
the wild silkworms of India and Africa[21] to the more esoteric Interestingly, nearly three decades passed before the first bio-
source represented by spider silk. The quest to commercialize medical use for a regenerated silk was reported in the patent lit-
spider silk, due to its favorable mechanical properties, seem- erature. In the 1960s, Bloch and Messores, of Ethicon Inc. (NJ,
ingly began with the inventions of Abbé Ramon de Termeyer in USA), were the first to propose the use of a LiSCN/LiBr recon-
the 18th century for his reeling device.[22] Over the years, these stituted silk as a replacement for the standard wax coating used
inventions were followed by others, such as those of the civil on silk sutures to reduce their limpness, fraying, and unwanted
war surgeon Burt G. Wilder.[23] The most successful attempts capillary action.[33] In the following years, while developments
are probably those made by the Madagascan spider silk industry, continued in the suture field, another two decades passed
which has produced, to date, only a handful of items destined before the first examples of nonfibrous silk-based biomate-
for the elite.[24] Yet, while producing arguably mechanically rial patents were reported. In 1986, a silk fibroin:fibrinogen
superior materials compared to those made of silkworm silk, glue, based on the “standard” LiBr reconstitution approach,
none of these endeavors were ultimately found to be scalable. was developed by a Japanese firm.[34] This was followed by the
Hence, given the coveted nature of silk, the fact that indus- first patent for a silk based porous scaffold in 1987, again from
trialists wished to replicate it may come as no surprise. In fact, Japan, produced from a freeze-dried native silk solution (i.e.,
nearly every single industrial fiber produced in the latter half silk extracted directly from the silk gland).[35] The 1990s saw
of the 19th and throughout the 20th century, from rayon to more patent applications from Japan, including powdered silk
nylon to Kevlar, has been developed in the hope that it would for wound dressings,[36] reconstituted silk films and molded
provide a suitable alternative to silk.[25] Nevertheless, even after gels for skin, blood vessel, and corneal coatings,[37] and colloidal
150 years of concerted research and development, and although silk for consumption in medicine.[38] However, in the 2000s,
replication of the properties and performance of silkworm silk an explosion occurred in the USA in research and commer-
is now possible, similar success with spider silk, and specifi- cialization activity around this area with the emergence of large
cally dragline silk, remains elusive. A complete overview of the patent families (>100) focused on the future medical exploita-
history, progress, and trends in artificial silk spinning from a tion of these materials.[39]
fiber performance perspective is available in a recent review In summary, looking back, the ability to unspin silk, and
that comprehensively covers this topic.[25] thereby reconstitute it, has been a monumentally disruptive
Fortunately, the attempts to replicate various silks resulted development in the field. It represents a platform technology
in several distinctly important innovations that led the bioma- for the development of biomimetic structures that are built
terials field to consider silk as more than just a fiber.[12] Akin with silk but are not built to replicate silk. While gaps still
to the ancient Chinese realizing that a silkworm cocoon can undoubtedly exist in our knowledge surrounding the process of
be unspun, early attempts at creating artificial silk led to the reconstitution and how this affects the integrity and application
conclusion that the silk fiber itself could be “unspun” back into of the silk proteins undergoing it,[40] the unspinning process
a processable protein feedstock, which could then be solidified has been widely adopted throughout the biomaterials field. This
into a variety of forms. According to the original patents, this is perhaps best evidenced by the impact of the landmark review
finding was largely motivated by a need to utilize the waste of Altman et al.[41] 15 years ago and the more recent protocol of
streams from the industry,[26] as the last tens to a hundred Rockwood et al.,[42] which leads us in the present day.
meters of silk from a cocoon could not be unraveled. (Today,
this would be labelled an exercise in sustainability.)
To the best of our knowledge, the first attempt to create an 3. Silk: Hierarchical and Crystal Structures
artificial silk feedstock appeared at the turn of the 20th century,
110 years ago. It began with the work of Baumann and Diesser, The fundamental building blocks of the silk biopolymer are
who proposed the dissolution of whole silk glands in formic amino acids that, through their sequence specificity and
subsequent secondary, tertiary, and quaternary structures, protein fiber is a composite material comprising a semi-crystal-
govern the protein’s overall function. In nature, silk and silkline silk core (i.e., silk fibroin), which is mainly responsible for
like proteins are made by several organisms such as spiders, the load-bearing capacity, and an outer layer of sericin, which
silkworms, scorpions, mussels, bees, and ants. However, the functions as a gumming agent.[45] However, emerging evidence
silk fibroins and silk-like proteins of each organism exhibit dif- suggests that sericin also inhibits the premature conversion of
ferent physical and biological characters due to their different soluble silk (silk I) into β-sheet-rich silk.[46]
amino acid sequences, spinning conditions, and hierarchical The Bombyx mori silk protein (i.e., silk fibroin) is very large
structures.[7b,43] The hierarchical structures of silk proteins vary and can be subdivided into light (≈26 kDa) and heavy (≈391 kDa)
among silk types.[43a,44] Silk proteins produced by spiders and chains that are linked by a single disulfide bond at the C-ter-
insects are referred to as silk spidroin and silk fibroin, respec- minus[47] (Figure 2). The C-terminal and N-terminal capping
tively. The term “silk fibroin” is commonly used to differentiate sequences are completely nonrepeating amino acid residues.
“virgin” silk (silk filament still encased by sericin) or silk cocoons The mechanical properties of silk fibroin arise due to the block
(i.e., the sericin-coated silk thread arranged into a cocoon) from copolymer-like arrangement of the silk heavy chain, which con-
purified silk (i.e., degummed; see Section 6). For the purpose of tains 11 short hydrophilic regions typically 31 amino acid long
this progress review, we will use the term silk fibroin to refer to and 12 hydrophobic blocks that account for 94% of the silk
degummed (Bombyx mori) silk unless otherwise stated. heavy chain. These hydrophobic blocks contain predominately
As discussed in the previous section, the silk fibroin of the glycine-X (GX) repeats, where X is alanine (A) (65%), serine (S)
domesticated silkworm (Bombyx mori) is the most well studied (23%), or tyrosine (Y) (9%).[47a] These GX blocks can be broadly
silk for biomedical applications due to its established supply classified into three groups: i) a highly repetitive GAGAGS
chain, abundance, and clinical track record. The Bombyx mori sequence that contributes to the bulk of the crystalline regions
Figure 2. Silk structure. A) Solution conformation of Bomyx mori silk. Hydrophobicity pattern of the heavy chain with possible chain folding and micelle
assembly of silk fibroin in water. Adapted with permission.[50] Copyright 2003, Macmillan Publishers. B) 2D silk fibroin schematic. Adapted with permis-
sion.[51] Copyright 2018, American Chemical Society. C) Primary structure of the Bombyx mori silk heavy chain. R01 to R12 and A01 to A11 represent
the arrangement of 12 repetitive and 11 amorphous regions, respectively. The approximate amino acid sequence of the R10 is shown by combination
of sequences of i, ii, and iii. Adapted with permission.[52] Copyright 2005, American Chemical Society. D) Hierarchal structure of spider silk. Adapted
with permission.[53] Copyright 2011, Elsevier. E) Primary structure of spider silks. Adapted with permission.[54] Copyright 2017, American Association
for the Advancement of Science.
and is typically found at the beginning of each motive, ii) a silks and the subsequent characteristics of silk fibroin as a
relative less repetitive sequence containing hydrophobic and/ biomaterial remains largely unexplored, despite the fact that crys-
or aromatic residues, namely, GAGAGY, GAGAGV, and GAGA- tallinity (i.e., the amount of crystalline region) affects the physical
GVGY, which form semi-crystalline regions, and iii) motifs and biological properties of silk-based biomaterials.
similar to i) except for the presence of an AAS motif, which
typically exists at the C-terminus of each motif and may play
a role for sheet-breaking.[48] Bombyx mori silk fibroin lacks the 4. Hydration State
tripeptide sequence arginine, glycine, and glutamic acid (RGD)
that is typically exploited by cells to mediated cell–substrate Silk and regenerated silk fibroin materials are expected to
attachment via integrin engagement; however, the N terminal exhibit high toughness and ductility because of the excel-
of the silk heavy chain contains a fibroblast growth-promoting lent mechanical characters of spider (dragline) silks found in
peptide.[49] Nevertheless, a sequence specificity exists between nature.[9,61] However, in addition to sequence specificity, the
different silkworm silks; for example, the Indian non-mulberry hydration state of silk is critical for its performance.[62] For
tasar silkworm (Antheraea mylitta) contains RGD sequences example, most native spider silks show significant fiber con-
that are absent in Bombyx mori silk. traction when transitioned from a dry state to a high humidity
Spider dragline silk, one of the toughest materials known to environment. Exposure to humidity facilitates the rearrange-
humankind, is composed of a skin layer and a bundle of micro- ment of the noncrystalline GPGXX sequence of orb web silks
fibrils (Figure 1).[5b,55] The microfibrils are composed of aligned and the glycine-glycine-X 310 helices in nonorbicularian species
granules, and their silk molecules form an amorphous phase (which lack the GPGXX sequence). This occurs due to disrup-
and β-sheet-rich crystalline regions.[5b] In both spider and silk- tion of hydrogen bonding by these sequences, which facilitates
worm silk fibers, the aligned β-sheet structure provides cross- the transition from a parallel arrangement for the fiber axis to
links between the β-sheet domains embedded in an amorphous a lower energetic configuration that is accompanied by fiber
matrix that consists of less orderly structures in the form of shrinkage and thickening.[63] Thus, water is a key component
random coils, helices, and β-turns.[56] These β-sheet crystals are that enables spiders to tailor the properties of their silks during
critical structures in the hierarchical structures of silk fibers, spinning and for in situ web tightening (a phenomenon also
because they play an essential role as cross-linking points and known as “supercontraction”).[63]
realize the stiffness, strength, toughness, and characteristic In nature, silkworm cocoons and spider webs/draglines
deformation behaviors.[56a,b,57] are tough structural materials that perform their function; for
The amino acid sequences that form the β-sheet are 7–9mer example, to capture prey in the spider’s web or to protect the devel-
alanine sequences for Nephila clavipes dragline silk and oping moth from predators and infection.[57b] The mechanical
GAGAGS for Bombyx mori,[58] whereas other silkworm silk spe- robustness of the native silk fiber has been exploited by humans
cies use polyalanine sequences to form the β-sheet structure.[6a] for biomedical applications both in preclinical (e.g., ref. [64])
The influence of the number of alanine residues on the sec- and clinical trials (detailed below). For example, silk fibroin scaf-
ondary structure and assembly behaviors of silk molecules has folds proposed for bone repair have shown a high compressive
been studied using wide angle X-ray crystallography as well as strength of ≈13 MPa when reinforced with Bombyx mori silk
solid-state nuclear magnetic resonance (NMR) spectroscopy. fibers.[64] A similar approach has been taken to enhance the
Those data from X-ray and NMR analyses reveal that short mechanical properties of Bombyx mori silk hydrogels for car-
poly(alanine) sequences, such as 6mers or shorter, form a tilage tissue engineering.[65] Recently, a high relative humidity
packed rectangular arrangement, while poly(alanine) sequences of >97% was found to cause a dramatic increase in the tough-
longer than 7mers pack in a staggered arrangement.[59] ness and crystallinity of silk films.[6a] This finding exemplifies
The β-sheet is the most fundamental secondary structure in how an appropriate hydration of silk molecules and materials
silk-based (bio)materials. The predominant β-sheet structure can achieve crystallization and plasticization simultaneously,
plays a key role in stabilizing silk materials via physical cross- resulting in a high-strength and tough silk material.
links, as the β-sheet behaves as a cross-linking point. Crystal
structures of silk β-sheets have been characterized using wide-
angle X-ray analysis. The crystal structure of Bombyx mori silk 5. Silk for Tissue Engineering and Drug
fiber has a unit cell with the space group P21-C22.[60] The crystal
Delivery—Expectations, Hopes, and the Reality
lattice of the Bombyx mori silk fiber reported by Marsh et al.
had unit cell dimensions of a = 9.40 Å, b = 9.20 Å, and c (fiber An excellent delivery system for bioactive molecules (e.g., small
axis) = 6.97 Å, while Takahashi et al. reported cell dimensions of molecular weight drugs, peptides, proteins, etc.) must meet
a = 9.38 Å, b = 9.49 Å, and c (fiber axis) = 6.98 Å.[60] The lattice of a number of requirements that include, but are not limited
other silks, such as Antheraea yamamai (Japanese silk moth), has to, biocompatibility, biodegradability, mechanical robustness
been characterized and reported by many groups.[57b] The unit and durability, and amenability to processing under ambient
cells contain four molecular chains, a pair of which symmetrically aqueous conditions that preserve the bioactivity of the payload.
forms a β-sheet structure via hydrogen bonds. The up-molecular Many of these requirements also apply for tissue engineering
and down-molecular chains also alternate with each other in an applications aimed at delivering or recruiting (endogenous)
antiparallel manner. Each silk has a different crystal lattice, which cells, although these silk constructs must also be able to pro-
can be attributed to differences of the silk amino acid sequences. vide cells with the necessary physical and biological cues to
However, the relationship between the crystal lattices of different achieve the desired function.
5.1. Silk Biocompatibility in conjunction with a notified body) before progressing the
device to first-in-man clinical assessment. Materials of animal
The exact set of biocompatibility requirements is application or allogeneic origin need to fulfill additional safety require-
specific, although many preclinical studies simply cite that silk ments (e.g., absence of infectious agents such as retroviruses,
can meet all the necessary requirements, or they make refer- etc.) before use in humans. However, from a regulatory per-
ence to silk as a “clinically approved” biomaterial for use in spective, Bombyx mori silk is regarded as a non-animal product
humans. However, this ignores our appreciation that a uni- (EU Council Directive 93/42/EEC, rule 17).
versal biocompatibility does not exist: a material needs to be Reports on the biocompatibility of silk in humans come pri-
fit for its intended use[66] (as documented by dedicated bio- marily from silk sutures (reviewed in ref. [41]) that have been
compatibility studies); thus, its performance is context specific. in use for several centuries[20] and from SERI Surgical Scaf-
The clinical approval of silk typically refers to its load bearing fold that obtained 510(k) clearance by the FDA in 2008 and
applications; degummed Bombyx mori silk fibers processed into underwent a market launch in 2013. Histological evidence of
a knitted surgical mesh (SERI Surgical Scaffold manufactured 69 breast tissue samples (by 60 patients) taken at stage 2 in
by Sofregen Inc., Medford, MA, USA), silk sutures (coated with patients undergoing two-stage breast reconstruction with SERI
waxes, Ethicon Inc. and several other manufacturers), and silk Surgical Scaffold showed a mild inflammatory response in 59
garments to treat dermatological conditions are in wide use patients, as confirmed by histology. This consisted of an infil-
today in the clinical setting. Therefore, their performance in tration of mostly macrophages and occasional multinucleated
humans is becoming better documented in the literature[41,67] giant cells that phagocytosed the silk fibers, as well as occa-
and is accompanied by a cadre of clinicians with experience sional lymphocytes and, rarely, neutrophils or polymorphonu-
working with these silk materials. clear cells.[73] Ordered collagen deposition was observed, with
Dedicated biocompatibility assessment is critical when gen- minimal or no encapsulation of the silk surgical mesh. These
erating novel silk formats (e.g., (nano)particles, hydrogels, scaf- clinical trial data[73] were similar to observations made in a
folds, films, coatings, etc.) to address areas of unmet clinical sheep study.[74] However, one patient had a postoperative hema-
need or when applying existing silk technologies to new indica- toma that led to mesh removal.[73]
tions. Any nonautologous material will elicit an initial foreign Synthetics are now the most widely used suture material,
body response that reflects the first steps of tissue repair.[68] but silk sutures are still in demand for specialized applications
Therefore, ensuring that the foreign body response is transient where exquisite handling is of paramount importance (e.g., eye
rather than chronic is a prerequisite to ensure that clinical end- surgery). Silk sutures are strong, are easy to handle, lie flat on
points can be met. Overall, biomaterial performance depends on the tissue surface, and allow for secure knots. Adverse reactions
the implantation site, size, geometry, surface topography, and to silk sutures are typically reported for virgin silk, where the
physical characteristics.[68] A systematic literature review[69] exam- silk filaments are still coated with sericin (and often with addi-
ining the performance of silk constructs (e.g., vascular grafts, tional waxes or silicones).[41] There is an ongoing debate about
ligaments, and wound dressings for skin grafting) in small and the potential role of sericin in these adverse reactions. However,
large animal studies overwhelming showed that a variety of dif- emerging evidence suggests that sericin on its own shows a low
ferent Bombyx mori silk constructs performed well across the allergenic and immunogenic profile in mice; in fact, this pro-
broad spectrum of indications and animal models.[69] Direct file is similar to that seen for silk fibroin or alginate.[75] These
in vivo comparison of silk with commonly used natural (e.g., observations are supported by in vitro data with macrophages:
collagen) and synthetic (e.g., polycaprolactone, polylactic acid, extracted sericin from Bombyx mori silk cocoons showed no
poly[lactide-co-glycolic acid]) biomaterials indicates that Bombyx significant release of the inflammatory marker TNF-α; sim-
mori silk fibroin is typically at least as good as these synthetic ilar observations were made with silk fibroin.[76] However,
materials and often superior than other natural biopolymers.[69] extracted sericin in combination with bacterial lipopolysaccha-
As new applications for silk emerge, appropriate biocompati- ride induced TNF-α release (but not for the silk fibroin group).
bility studies must be performed to support these developments. Furthermore, recoating of silk fibroin with sericin showed no
For example, silk nanoparticles for anticancer drug delivery are macrophage response, while virgin silk induced a high level
typically designed for intravenous administration[70] and thus of TNF-α release.[76] These data suggest that other leachable
require hemocompatibility assessments because biological compound(s), or these compounds combined with sericin,
performance cannot be deduced by extrapolating results from may be responsible for the adverse clinical reactions reported
macroscopic films[71] to nanoscale particles.[72] An initial proof of for silk.[69,76] For example, patients subjected to bilateral cata-
biocompatibility is a first step to translate silk technologies from ract surgery showed no suture reaction on the first eye but a
the bench to the clinical setting. For example, regulatory frame- severe reaction on the second eye when it was treated six to
works imposed by the Pharmaceuticals and Medical Devices three months later. This suggested that these patients had
Agency Japan, the Food and Drug Administration (FDA, USA), undergone a sensitization toward virgin silk. Prompt removal
the Medicines and Healthcare Products Regulatory Agency of the offending silk suture resulted in significant clinical
(UK), and the European Medicine Regulatory Agency (EU) for improvement.[77] Examining the clinical literature regarding
medical devices (e.g., Regulation (EU) 2017/745 to obtain CE silk sutures and identifying the exact cause of the adverse reac-
marking analogous to the Class III Premarket Approval/510(k) tion are challenging because often little (or no) information is
in the USA, and the Australian Register of Therapeutic Goods provided about the exact nature of the silk suture (e.g., virgin
certificate of inclusion) stipulate that a biological safety assess- silk, type of coatings, etc.). Nonetheless, an allergic response to
ment needs to be conducted first (by an ISO certified laboratory, Bombyx mori virgin silk is documented for the occupational and
domestic setting (reviewed in ref. [78]). For example, exposure to tissue was mechanically strong.[74] Clinical hernia repair in a
virgin silk fibers and repurposed silk waste (e.g., silk floss incor- horse showed incomplete SERI Surgical Scaffold degradation at
porated into rugs and bedding) has been linked to the develop- 2 years postimplantation, but no hernia relapse.[88]
ment of asthma in silk weavers[79] and children,[80] mounted by The time scale for silk degradation depends on a number of
an IgG and IgE immune response.[81] Textile workers have an factors, including, but not limited to: i) the amount of material, ii)
increased risk of developing chronic obstructive pulmonary dis- gross morphology, iii) silk secondary structure, iv) silk treatment
ease, and this risk is highest in silk workers.[82] history, v) mechanical environment, and vi) implantation site
Complete and reproducible sericin removal from Bombyx (or final destination). The implantation site directly impacts the
mori silk (a.k.a. degumming) is an essential step in silk utiliza- type of proteolytic enzyme encountered by the silk, because these
tion. Clinically acceptable limits for residual sericin levels for enzymes vary between tissues, cells, and subcellular location.
marketed silk products have not been released into the public Silk fibroin sequence alignment indicates a susceptibility
domain (note that SERI Surgical Scaffold is described by the to a number of proteases (e.g., protease XIV, α-chymotrypsin,
manufacturer as highly purified silk with ≥95% purity). Cur- proteinase K, papain, matrix metalloproteinases, collagenase,
rent evidence from both preclinical in vivo studies and clinical etc.)[51,89]). Nonetheless, predicting silk fibroin degradation simply
experience in humans across a range of applications indicates based on the primary sequence is unreliable; for example, chymo-
that Bombyx mori silk fibroin is biocompatible, provided that all trypsin has 434 cleavage sites in the silk heavy chain and 81 in the
other contaminants are successfully removed. light chain, while protease XIV has 348 in the heavy chain and 41
Sericin has traditionally been linked to the adverse effects in the light chain. Despite numerous cleavage sites, chymotrypsin
reported for virgin silk (reviewed in ref. [78]). However, over the treatment for 20 days had no quantifiable effect on silk fibroin,
past decade, sericin has emerged as an interesting biopolymer while protease XIV significantly degraded silk fibroin in vitro.[51]
(reviewed in ref. [83]), and dedicated biocompatibility studies are Papain, a cysteine protease enzyme that mimics the activity of
now showing encouraging results in relation to the allergenic lysosomal enzymes, has 26 cleavage sites in the silk heavy chain
and immunogenic profile of sericin (e.g., ref. [75]). An increasing (albeit exclusively in the amorphous regions) and 15 in the light
number of studies report the biomedical use of the biopolymer chain, and it caused significant silk fibroin degradation over 20
sericin. For example, the development of composite sericin/ days but at a slower rate than protease XIV. Similar observations
silicone nerve guides[84] or sericin/polyacrylamide hydrogels pro- were made with isolated lysosomal enzyme preparations.[51]
posed for dermal repair.[85] Preliminary Phase I clinical trials using Overall, these studies exemplify that the structure beyond
sericin composite wound dressings for split-thickness skin grafting the primary sequence is of critical importance for silk degra-
are on going and the results are eagerly awaited (NCT01539980 dation. The current working model supports the notion that,
and NCT02643680 reported at www.ClinicalTrials.gov). for Bombyx mori silk, degradation begins with the 11 hydro-
philic amorphous segments in the silk heavy chain, as well as
the C-terminal and N-terminal and the silk light chain, which
5.2. Silk Biodegradation consist of completely nonrepeating amino acid sequences;
this is then followed by degradation of the more crystalline
Silk sutures are classified by regulatory agencies as non-biode- sequences.[51,61c,89] The tightly packed crystalline domains are
gradable because regulatory guidelines expect a loss of most degraded last.[90] Furthermore, the silk format is a critical factor
tensile strength within 60 days postimplantation. Over this time in determining degradation rates, as in vivo studies in rodent
scale, silk sutures do not lose their mechanical performance, models indicated faster degradation for open silk structures
as they require longer time frames to degrade in humans.[41] than for tightly packed monolithic silk fibroin films (rank order:
In patients undergoing two-stage breast reconstruction, histo- hydrogel > silk scaffold > monolithic film).[69]
logical evidence of breast tissue samples taken from 60 patients Protease XIV is a useful model enzyme for studying silk degra-
at stage 2 (median 152 days after initial scaffold implantation, dation and for comparing with earlier studies. However, protease
range 74 to 357 days) showed consistent SERI Surgical Scaffold XIV is a nonmammalian enzyme cocktail, so it must not be used
degradation (although this was not quantified). The one excep- to deduce or predict biocompatibility performance. Silk, as a pro-
tion was a patient that had a postoperative hematoma, which tein-based biopolymer, is commonly considered to yield harm-
was accompanied by an apparent lack of silk degradation.[73] less biodegradation products; however, a more critical inspec-
The silk protein is known to degrade in vitro and in vivo in tion of silk and its degradation products is timely. For example,
response to proteolytic enzymes,[69] as exemplified by studies silk fibrils have molecular-level similarity to amyloid fibrils,[91]
with silk films (e.g., ref. [86]) and porous silk scaffolds (e.g., and they were also reported to enhance amyloidosis of amyloid
ref. [87]). Experience with SERI Surgical Scaffold in a sheep protein through a mechanism based on cross-seeding effects.[92]
model of two-stage breast reconstruction showed progressive However, when silk nanofibrils and microfibrils were composed
degradation and vascularization of the silk mesh: at 1 month of β-sheets, which are known to affect various properties of silk
postimplantation, tissue ingrowth and marked vascularization fibers, they demonstrated no significant cytotoxicity toward in
were evident; at 4 months, the mesh was no longer felt through vitro neuronal cells. When the silk fibroin was degraded with
the skin; and at 12 months, the mesh degradation and vascu- chymotrypsin to yield mainly unordered soluble fragments with
larization were scored as mild but with substantial silk loss that a low β-strand content, the degradation products caused no sig-
precluded mechanical testing of the remaining SERI Surgical nificant amyloidosis. By contrast, significant cytotoxicity was
Scaffold.[74] At 12 months, the SERI Surgical Scaffold had stim- observed when silk fibroin was degraded with protease XIV due
ulated extensive type I collagen deposition and the resulting to the formation of soluble β-sheet rich fragments.[61c]
Formation of β-amyloid structures is a concern because amy- Serica Technologies Inc. was subsequently acquired by Allergan
loid beta fibrils are a hallmark of Alzheimer’s disease.[61c,93] Inc., and the SERI Surgical Scaffold became commercially avail-
Preliminary studies in mice injected with self-assembling silk able for soft tissue support and repair in 2013 and was since
fibroin hydrogels into the caudate putamen (striatum) showed then acquired by Sofregen Medical Inc. (Medford, MA, USA).
no decline in cognitive function or animal behavior over the The current SERI Surgical Scaffold indications are for
6 week study period.[94] abdominal wall reconstruction[98] and investigational plastic sur-
gery applications, including total body contouring, brachioplasty,
abdominoplasty, mastopexy, and breast reconstruction (Table 1).
6. Processing of Silk Cocoons—Generating The clinical performance of SERI Surgical Scaffold has been
reported in the literature, which includes open label clinical
Silk for Biomedical Use
trials and case reports (Table 1). Many of these encouraging
Unspinning the Bombyx mori silk cocoon and degumming to clinical studies have been sponsored by Allergan Inc. A few
remove sericin are two crucial steps that yield silk suitable for independent retrospective clinical reports of small patient
biomedical use. Sericin can be removed by enzymatic methods cohorts are reporting side effects (e.g., poor scaffold integration,
(i.e., digesting sericin but not silk) or chemical processing (e.g., see Table 1, Figure 3), often requiring surgical removal of the
alkaline treatment). The latter approach is widely used and typi- mesh.[67b,105,106] Therefore, some clinicians are abandoning the
cally involves boiling silk in sodium bicarbonate for 20–60 min.[42] use of SERI Surgical Scaffold in their clinical practices,[105] and
Degumming times as short as 5 min are also sufficient to remove caution has been raised by others.[108] In 2013, Allergan Inc. vol-
sericin while minimizing silk damage, which usually occurs due untary withdraw several SERI Surgical Scaffold batches due to
to cleavage of the disulfide bond between the silk heavy and lights concerns about product sterility. How this might have affected
chain and fragmentation of the amorphous silk sequences in the the reported adverse events is not known.
silk heavy chain, which results in polydispersed silk.[95] We are familiar with silk for the textile industry, although
The degummed silk fibers can be fully reverse engineered by silk garments are also used clinically to treat dermatological
dissolving them in a high concentration chaotropic agent (for conditions, especially atopic dermatitis[109] and acne vulgaris[110]
example, 9.3 m lithium bromide) at 60 °C over several hours (Table 2). Mechanical skin irritation by harsh, rough (e.g., wool),
to dissemble the higher order silk structure. The resulting and short (e.g., cotton) textile fibers is thought to contribute to
silk fibroin solution is then dialyzed extensively against water atopic dermatitis. Furthermore, the skin of atopic dermatitis
to yield an aqueous silk solution that is stable at room tem- patients is often colonized with Staphylococcus aureus and the
perature for weeks and at 4 °C for several months.[42] When extent of colonization correlates with the severity of the disease.
compared to native silk feedstock, this reverse engineered silk Silk fibers are very long (up to 1500 m) and smooth, so they
fibroin solution has a reduced solution conformation[96] and minimize mechanical irritation when knitted into clothing.
changed rheological properties.[40d] This silk clothing has been chemically modified to achieve
The reverse engineered aqueous silk fibroin solution is com- antibacterial properties with the aim of reducing Staphylococcus
monly used to generate novel silk formats; for example films, aureus colonization of the skin. Sericin-free silk has also been
fibers, scaffolds, and (self-assembling) silk hydrogels, as well as covalently functionalized with 3-trimethylsilylpropyl-dimeth-
(nano)particles and (nano)coatings, and these formats are often yloctadecyl ammonium chloride (AEM 5700/5772; AEGIS),
achieved using an all aqueous processing under ambient condi- resulting in commercial products (e.g., DermaSilk) for the
tions. These mild processing conditions are ideal for preserving treatment of atopic dermatitis. These silk garments use highly
the activity of biologics. purified silk to minimize the risk of contact dermatitis.[111]
The silk garments are also knitted in a specific fashion to
improve transpiration of sweat through the fabric (unlike eve-
7. Present Routine Clinical Use of Silk ryday silk, which can worsen atopic dermatitis by trapping
moisture). A randomized double-blind study in 30 patients with
The silk surgical mesh SERI Surgical Scaffold, silk sutures, atopic dermatitis on both arms received an AEM 5700/5772
and silk clothing to treat dermatological conditions are the only functionalized silk sleeve and a silk-only sleeve. Patients treated
available products in routine clinical use today. All these prod- with the silk sleeve showed a rapid improvement within 2 weeks
ucts are manufactured by unwinding Bombyx mori silk cocoons but remained similar until the end of the study. The contralat-
and working with the silk thread. The clinical performance of eral arm treated with the AEM 5700/5772 functionalized silk
silk sutures, their adverse effects, and the developments and showed similar results at 2 weeks but reached a greater level of
potential solutions to improve suture performance have been improvement over 4 weeks.[109d] Other clinical trials using AEM
reviewed previously.[41] 5700/5772 functionalized silk garments in small patient cohorts
The SERI Surgical Scaffold technology is based on work con- reported substantial improvements in skin conditions (Table 2).
ducted by David Kaplan and co-workers at Tufts University, Med- By contrast, a randomized, controlled, observer-blinded clinical
ford, MA, USA.[67a,97] The resulting patent portfolio and proprie- trial in 300 children showed only a 3% reduction in skin infec-
tary silk processing technologies formed the basis of the spin-out tion compared to control and was therefore not regarded as
company, Serica Technologies Inc. (Medford, MA, USA). Serica providing a significant clinical benefit.[109e] Overall, these clin-
Technologies Inc. was able to prove to the FDA that SERI Sur- ical trials are difficult to conduct, and the use of different silk
gical Scaffold was “substantially equivalent” to existing surgical garments (DermaSilk and DreamSkin, see Table 2 for details)
meshes and thus received 510(k) clearance to market the device. undermines the power of the study. Furthermore, the selection
Table 1. Published data reporting the clinical use of SERI Surgical Scaffold in humans.
Year Article type Patient Study sponsor Intervention Clinical follow up Reported outcome Reference
number (months)
2013 Retrospective case report 1 Allergan Abdominoplasty and use of scaffold to provide 24 Contour and flatness of the anterior abdominal wall was maintained [99]
soft-tissue support to the abdominal fascia in
patient with massive weight loss
2014 Retrospective study 141 Allergan Revision of breast augmentation (n = 40); revision of 0 to 12 Adverse side effect reporting voluntary [100]
Multi center breast reconstruction (n = 24); mastopexy augmen- Surgeons rated the ease of use a mean of 2.86 (scale 0–3)
www.advancedsciencenews.com
tation (n = 20); mastopexy augmentation-revision Surgeons rated their satisfaction a mean of 9.31 (scale 0–10)
Adv. Healthcare Mater. 2018, 1800465

(n = 16); hernia repair (n = 11); other (n = 30)
2014 Prospective study 139 Allergan 2-stage implant-based breast reconstruction 6 75 subjects undergone stage 2, subject satisfaction score 4.3 ± 0.91 [101]
Multi center (5 best). Investigator satisfaction score was 9.4± 0.84 (10 best). Adverse
effects in 214 breasts: tissue necrosis (6.1%), seroma (6.1%), hematoma
(2.8%), breast infection (1.9%), cellulitis (1.9%), implant loss (1.9%),
capsular contracture (0%)
2014 Retrospective case report 1 Allergan Abdominoplasty and lower body lift of in patient 7 No complications reported, improve patient satisfaction [102]
with massive weight loss. Scaffold implantation on
left lower body only
2014 Retrospective case report 1 Allergan Brachioplasty 6 No complications reported, perceived faster maturation process and a [103]
better-quality scar
2014 Retrospective study 172 Not disclosed 77 patients (71 women, 6 men) underwent 18.4 ± 7.5 The overall complication rate (N77) was 6.5%, consisting of 2 wound [98]
Multi center abdominal wall fascial repair or reinforcement. The dehiscences, 1 with device exposure, 1 seroma, 1 infection with explanta-
remaining 95 patients not reported on tion, and a perioperative bulge requiring reoperation
2014 Retrospective study 15 No Direct-to-implant after skin-sparing mastectomy 6 to 13 Capsular contraction (35%); loss of scaffold due to necrosis (n = 1); [104]
1800465 (9 of 26)
Single center seroma (n = 1); hematoma self-limiting (n = 1); patient satisfaction
(5.77 out of 10)
2015 Correspondence 5 No Unilateral skin-sparing mastectomy and imme- Not reported Late infection (6 weeks and 3.5 months postsurgery) in 2 breasts leading [105]
Retrospective study diate reconstruction to scaffold and implant removal. In 2 patients successful completion
Single center with tissue integration and vascularization
2015 Prospective study 139 Allergan 71 patients undergoing 2-stage breast 12 Investigator satisfaction score was 9.4 ± 0.91 (10 best) and patient [73]
Multi center reconstruction scores was 4.5 ± 0.82 (5 best). Complication rates in 105 breasts were
tissue necrosis (6.7%), seroma (5.7%), hematoma (4.8 percent), implant
loss (3.8%), capsular contracture (1.9%), breast infection (1.0%)
2015 Correspondence 4 No Breast reconstruction 12 Late infection with Ps. aeruginosa in 2 patients at 5 months resulting in [106]
Retrospective study implant replacement. Lack of mesh integration (or degradation) in all 4
Single center patients
2017 Prospective study 103 Allergan 2-stage implant-based breast reconstruction 24 Investigator satisfaction high [107]
Multi center
2018 Prospective study 16 No Direct-to-implant reconstruction with surgical scaf- 24 to 37 No intraoperative complications. Adverse effects in 22 breasts: Postopera- [67b]
Single center fold after skin-sparing mastectomy tive bleeding, that required reoperation occurred in 5% breast, postopera-
tive seroma in 45% and surgical site infection in 9%. Scaffold-related
complications occurred in 14% breasts, lack of scaffold integration in all,
resulting in skin ulceration in 2 and the scaffold lying free in the breast
pocket surrounded with seroma in one
© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

www.advhealthmat.de
of water that acts as a plasticizer) and formed

an effective barrier against bacterial infection
in vitro. The films were found to be biocom-
patible for their intended use according to
ISO 10 993 tests for the biological evaluation
of medical devices. In a rabbit full-thickness
wound healing model, healing was three days
faster in wounds treated with silk films than
in wounds treated with Suprathel, a poly
urethane-based synthetic wound dressing,
and seven days faster than in wounds treated
with the Sidaiyi wound dressing or phos-
phate buffered saline (PBS) treated controls
(Figure 4). Silk-film-treated wounds showed
the development of an organized epidermis
by day 14 post-treatment and showed a mature
and organized collagen matrix, hair follicles,
and blood vessels histologically by day 21.[113]
These results were further verified in a por-
cine full-thickness wound healing model prior
to initiation of a Phase I clinical trial that ran
from August 2013 to September 2014. This
clinical trial enrolled 71 patients (36 randomly
Figure 3. Examples of SERI Surgical Scaffold implant loss in humans. A) Silk fibroin surgical
mesh prior to implantation. B) Intraoperative view showing a free lying scaffold in the breast assigned to a silk film group and 35 to a Sid-
pocket. C) Retrieved scaffold surrounded with seroma. D) Interaoperative view of surgically aiyi group). The silk-based wound dressings
removed scaffold with interpenetrated granulation tissue/scar plate (at > 5 months), and were used to cover donor sites following sur-
E) histology of retrieved sample showing granulation tissue with neutrophiles and giant cells gical harvesting of split-thickness skin grafts;
at the material (1) interface (dotted line). (B,C) Reproduced with permission.[67b] Copyright healing was significantly faster in the silk film
2018, Elsevier. (D,E) Reproduced with permission.[106] Copyright 2015, Elsevier. group than in the Sidaiyi wound dressing
group and 100% of the wounds were healed by
of patients with only moderate eczema (typically not treated day 14 postinjury in the silk film group. One case of inflamma-
with these types of garments) and the often limited wearing of tory reaction to the silk film was noted but the exact etiology was
the garments are not in line with the clinical recommendations. not determined. No cases of wound exudation were observed,
Therefore, these difficulties might result in underestimation indicating that the silk films maintained a clean wound environ-
of treatment effects. Overall, improving these silk garments to ment with suitable moisture levels. The films showed good adhe-
maximize their clinical performance warrants more research. sion to the wound surface and no changes of the wound dressing
were required. As the wound healed, the silk films spontaneously
detached from the regenerated skin areas. The exact mechanism
8. Clinical Trials Using Silk underlying the improved clinical performance observed for the
silk film group over the Sidaiyi group is currently unknown.
The renewed interest in silk for biomedical use over the past However, this study demonstrated the ability to manufacture
20 years has resulted in a number of clinical trials; however, silk films under Good Manufacturing Practice requirements
historically, these data sets have been difficult to source. Since and their successful use as wound dressings for skin repair and
2007, the FDA has mandated that drug and device manufac- regeneration.[113] The potential for relatively easy modification
turers register clinical trials (www.ClinicTrials.gov) (Table 3). of silk films for additional functionality, such as the incorpo-
Silk-based biomaterials show particular promise for skin ration of pores or the introduction of bioactive molecules,[114]
wound healing due to their hemostatic properties, low inflam- makes silk films particularly attractive as wound dressings.
matory potential, and permeability to oxygen and water, as Thin silk films have also been used in prospective human
well as their ability to function as a barrier to bacterial coloni- clinical trials to repair acute and chornic tympanic membrane
zation (Table 3). Sidaiyi, a silk fibroin sponge attached to a sili- perforations.[115] These silk patches (Tympasil, Daewoong-Bio,
cone membrane, is a first generation wound dressing currently Seoul, South Korea) were generated using reverse engineered
approved by the China Food and Drug Administration for clinical Bombyx mori silk fibroin. The process leading to stabilization
use in that country.[113] The Sidaiyi platform was first compared of these silk patches has not been established, but physical
to silk fibroin films for wound healing applications in preclinical cross-linking is most likely because the brittle patches were first
animal models, followed by a randomized, single blinded Phase I wetted in PBS to plasticize them to facilitate their trimming to
clinical trial. Silk films were made by casting an aqueous Bombyx the required size and surgical placement.
mori silk fibroin solution in a mold and treating it at 65 °C and The first clinical trial involved 52 patients with acute trau-
90% relative humidity for 100 min. The resulting 64.9 µm films matic tympanic membrane perforation who were treated with
were water-resistant (albeit their ability to bind a small amount either a silk film or a paper-based membrane.[115b] A number of
Table 2. Published data reporting the clinical use of silk garments for the treatment of atopic dermatitis in humans.
Year Article type Patient Study sponsor Patients and intervention Clinical Reported outcome Reference
number follow up
2004 Retrospective study 46 ND Children 4 months to 10 years. DermaSilk 1 week Local score of treated and untreated [109c]
Single center treatment or cotton clothing, topical moistur- area of same child (SCORAD index)
izing cream or emulsion Overall DermaSilk decreased atopic
dermatitis severity and local improve-
ment of silk treated areas
2006 Retrospective study 15 ND Children 0.6–9.2 years, body suits made out 3 weeks Significant improvement of atopic [112]
Single center of DermaSilk and cotton (50:50). Cotton side dermatitis by assessing eczema area
also received topical corticosteroid and severity (EASI index) irrespec-
tive of intervention. No significant
difference between DermaSilk and
corticosteroid treatment
2007 Retrospective study 22 ND Children 5 to 12 years old. DermaSilk, silk 3 months DermaSilk arm significantly improved [109b]
Single center, random- fabric, and cotton. Control on contralateral arm over study period when compared to
ized, single-blinded (silk for first for 2 weeks followed by cotton cotton. Silk control treatment showed
for rest of study period), topical moisturizing significant improvement of SCORAD
cream or emulsion, antihistamines prn index
2008 Retrospective, random- 30 ND Age 3 to 31 years (mean 14.2). DermaSilk 1 month Significant decrease in SCORAD index [109d]
ized, double-blinded sleeve versus equivalent silk only fabric for both groups. Silk only fabric rapid
reduction over 2 weeks only while
DermaSilk over 4 weeks; decrease in
pruritus values similar during first 2
weeks but further decrease for Der-
maSilk group until end of study
2013 Retrospective, single 22 ND Age 4 to 18 months. Acute phases treated as 24 months Significant reduction in topical [109a]
center, double blind ran- per international guidelines. DermaSilk body steroid use for DermaSilk group;
domized controlled trial and tights, or wear clothes in pure cotton; subjective pruritus also reduced
expect May to September significantly
2017 Retrospective, multi- 300 University of Not- Children aged 1 to 15 years. Standard care 24 weeks No statistical difference between [109e]
center parallel-group, tingham, National or standard care plus silk garment (either groups for eczema area and severity
randomised, controlled, Institute for Health DermaSilk or DreamSkin). Eczema outcome (EASI index). Less frequent skin infec-
observer-blind trial Research Clinical and skin infections tions in silk group. Data not stratified
Research Network for different silk garments. Included
cost-benefit analysis rejects garments
SCORAD: An atopic dermatitis scoring system combining extent, severity, and subjective symptoms. EASI: Eczema area and severity index. DermaSilk: AL.PRE.TEC. S.r.l.
Italy, Silk is covalently functionalized with the antimicrobial 3-trimethylsilylpropyl-dimethyloctadecyl ammonium chloride (AEM 5700/5772; AEGIS). DreamSkin: DreamSkin
Health Ltd UK. The silk fabric contains zinc-based antimicrobial and is coated with 2-(methacryloyloxy)ethyl-2-(trimethylammonio) ethylphosphate-stearylmethacrylate
copolymer to from multilayer lamellar structures. The amphiphilic copolymer is proposed to serve as both a barrier and moisturizer.
conservative treatment modalities had been explored to support two treatment groups. However, the surgical time for the silk
the (often spontaneous) healing of an acutely perforated tym- patch was very short (13.7 vs 29.5 min) and no sourcing of con-
panic membrane, including the placement of a “patch” on top nective tissue was required for the graft.[115a]
of the damaged tympanic membrane. In this trial, the silk or
paper patches were surgically placed and removed 7 days later,
9. Preclinical Use of Silk—The Future
when the tympanic membrane appeared fully regenerated. The
closure rate was similar for both the silk film and paper mem- As we move from the routine clinical use of silk fibers to human
brane (92.3 and 84.6%, respectively), but the silk patch signifi- clinical trials, the line between a silk thread-based medical device
cantly shortened the healing time from 16.7 to 13.7 days. and other forms of silk starts to blur. A very wide spectrum of silk
A similar improved healing with a silk patch was also pre- materials and formats is now emerging in preclinical studies.[167]
viously reported in animal studies.[116] A follow-up study of 40 We will first review silk solutions derived from reverse engineered
patients with chronic traumatic tympanic membrane perfora- Bombyx mori silk, followed by more complex formulations.
tion showed that patients treated with a silk patch (Tympasil)
had lower otorrhea, minor complication rates, and high patient
satisfaction when compared with conventional perichon- 9.1. Silk Solution
drium myringoplasty.[115a] The silk and autologous patches
were removed one week after placement, and the postoperative Bombyx mori silk in its solubilized aqueous form has been
hearing outcomes were not significantly different between the investigated for a range of therapeutic applications, including
Table 3. Reported human clinical trials using silk.
Title Status Number Study Study results Primary outcome measure Identifier
of patients completion date
SeriACL device trial for anterior Unknown. Status 30 October 2008 Not posted Safety—measured by device related NCT00490594
cruciate ligament (ACL) was active not serious adverse events (time frame:
reconstruction recruiting 12 months)
Coated VICRYL Plus suture compared Completed 101 May 2009 Posted Mean score on cosmetic outcome visual NCT00768222
to Chinese silk in scheduled breast analog scale (time frame: 30 days (+/−5)
cancer surgery postoperative)
Clinical and economic outcomes Withdrawn (strategic ND August 2014 Not posted Incidence rate of implant loss (SERI NCT02033590
with the use of SERI Surgical priorities impacted Surgical Scaffold and breast implant)
Scaffold in direct-to-implant breast study) (time frame: 52 weeks)
reconstruction
Evaluation of HQ Matrix medical Completed 71 September 2014 Posted Time to wound healing (time frame: Days NCT01993030
wound dressing for healing of donor 0, 3 ± 1, 7 ± 1, 10 ± 2, 14 ± 3, and 21 ± 3
site wounds postoperation)
SERI Surgical Scaffold use in Completed 100 February 2015 Posted Investigator satisfaction following use of NCT01389232
reconstruction postmarket study for SERI Surgical Scaffold evaluated using an
tissue support and repair in breast 11-point scale questionnaire (time frame:
reconstruction surgery in Europe six months)
Efficacy and safety of silk fibroin with Completed 29 May 2015 Not posted Clinical efficacy of wound dressing con- NCT02091076
bioactive coating layer dressing taining silk fibroin with a sericin bioactive
coating layer dressing in the treatment of
split-thickness skin graft donor sites (time
frame: within 14 days)
SERI Surgical Scaffold postmarket study Completed 17 March 2016 Not posted Incidence of Implant Loss (time frame: NCT01914653
of soft tissue support and repair in 24 months postoperatively)
breast reconstruction
A SERI Surgical Scaffold postmarket Completed 1 June 2016 Not posted Rate of hernia recurrence NCT01981044
study of soft tissue support in ventral
hernia repair
Circumferential periareolar mastopexy Completed 14 June 2016 Not posted Size of the areola at 12 months NCT02293798
using SERI Surgical Scaffold as measured by physical mammometry
Suture materials: an evaluation Completed 36 November 2016 Not posted Accumulation of soft deposits NCT03410433
(time frame: 7 to 14 days)
SERI Surgical Scaffold Support of the Completed 76 November 2016 Posted Nipple to fold measurement on stretch NCT02016612
lower pole of the breast (SeriSupport) (time frame: 1 year post op)
Evaluation of HQ Matrix soft tissue mesh Unknown 144 December 2016 Not posted Postoperative recurrent rate NCT02487628
for the treatment of inguinal hernia (time frame: day 1 postoperation)
The comparison of microbial adher- Unknown. Status 30 May 2017 Not posted Bacterial counts on blood agar plates NCT02653924
ence to various sutures in patients was not yet recruiting from each suture will be quantified in CFU
undergoing oral surgery (colony-forming units) and expressed as
total bacteria/suture (time frame: outcome
measure will be assessed 10 days after
sample incubation for the different sutures
obtained from each study participate)
Initial safety evaluation of FibroFix Terminated (Safety). 4 July 2017 Not posted Safety (time frame: 12 months) NCT02205645
Meniscus Devices explanted.
12 m postexplant
safety f/u as agreed
with UK MHRA)
DACCa) in the REduction of Surgical Recruiting 712 December 2018 Not posted 30 day infection rate (time frame: 30 days) NCT02992951
Site INfection (DRESSINg)
Porous tissue regenerative silk scaffold Not yet recruiting 120 January 2021 Not posted Performance analysis of meniscal scaffold NCT02732873
for human meniscal cartilage repair (time frame: At 12 months follow up)
(REKREATE)
a)Although
not silk fibroin, dialkylcarbamoylchloride (DACC) is found as a hydrophobic coating on spider webs.
the treatment of ocular conditions, including dry eye[120] and

corneal injuries.[5b] Blindness from corneal disease affects over
50 million people worldwide, while another 337 million people
suffer from dry eye disease, representing a significant health-
care burden.[5b,121]
For example, silk fibroin treatment resulted in increased tear
production and reduced the corneal irregularities observed in
the absence of treatment in a mouse dry eye model (consisting
of NOD.B10.H2b mice exposed to desiccation stress and scopol-
amine hydrobromide treatment for 10 days). Silk fibroin treat-
ment inhibited detachment of corneal epithelial cells, increased
the number of conjunctival goblet cells, and inhibited the secre-
tion of inflammatory factors in the lacrimal gland of the eye,
resulting in recovery of the tear film and mucus layer of the eye,
improved corneal health, and reduced dry eye symptoms. Other
anti-inflammatory agents, such as cyclosporine and corticoster-
oids, are available on the market for the treatment of dry eye,
but silk has demonstrated a potential multitarget therapeutic
effect that lacks the common side effects, such as pain and
irritation, and other complications associated with long-term
corticosteroid use.[44] Silk fibroin was demonstrated to stabilize
the tear film through anti-inflammatory effects in the lacrimal
gland and increased number of conjunctival goblet cells, but the
mechanisms underpinning these observations are unknown.
Clinical approaches to treat corneal injuries are relatively
limited and predominantly involve topical application of anti-
inflammatory or antimicrobial agents that do not promote
tissue regeneration. A rabbit corneal injury model, which
involved removal of a 7 mm diameter section of the central cor-
neal epithelium, was used to study the effects of an aqueous
Bombyx mori silk fibroin solution (deemed “silk-derived pro-
tein” due to an additional autoclaving step during solubilization
in lithium bromide that results in a heterogeneous population
of low molecular weight silk fragments) on corneal epithe-
lial healing.[5b] All treatments showed corneal wound closure
by 48 h postinjury, as indicated by fluorescein staining; how-
ever, treatment with silk accelerated the rate of wound healing
threefold in the first 6 h postinjury. Relative to a PBS-treated
control, silk treatment resulted in a significant increase in the
numbers of proliferating Ki-67 positive epithelial cells, a dose
dependent increase in epithelial cell attachment to the under-
lying basement membrane (as indicated by focal adhesion
kinase staining), and a dose-dependent reduction in MMP-9,
Figure 4. Silk fibroin films for wound healing applications. A) Healing a metalloprotease involved in matrix remodeling and corneal
of rabbit full-thickness wounds over a 21-day period following appli- repair. Finally, compared to the PBS-treated control, the silk-
cation of Bombyx mori silk fibroin films, a polyurethane based wound
dressing Suprathel, a silk-silicone wound dressing Sidaiyi, and a blank
treated group showed the formation of epithelial layers with
control treated with PBS. Silk-film-treated wounds healed 3 days faster tight junctions (ZO-1 staining) that more closely resembled
than Suprathel-treated wounds and 7 days faster than Sidaiyi-treated and those of healthy corneas.[5b] The potential of the silk solution
untreated wounds. B) Histological evaluation of the wounds: A moderate in aiding wound healing is clearly demonstrated, but the exact
to complete epidermal organization in silk film treated wounds by day 14 mechanism of action is yet to be determined.
and mature regenerated tissue with well-formed collagen matrix by day
21. C) Photographs of representative silk-film-treated partial thickness
wounds in a human Phase I clinical trial, showing complete healing at day
11, with an average time for complete wound healing of 9.86 ± 1.97 days 9.2. Silk Films
(n = 36). D) Kaplan–Meier curves comparing cumulative healing by treat-
ment group. Reproduced with permission.[113] Copyright 2017, Wiley-VCH. Silk films are among the most extensively explored biomaterials
due to the ease of their fabrication and characterization and
treatment of diabetes,[117] chronic wounds,[118] and inflamma- their versatility. Silk films have been explored for their potential
tion.[119] Recent studies have investigated the utility of regen- in drug delivery.[61b] wound healing,[113,114,122] corneal replace-
erated silk fibroin solution in preclinical animal models for ment, and flexible electrode[123] applications, among others.
Due to their transparent nature, silk films

have been particularly well explored for
ocular applications, including corneal and
retinal regeneration. Silk films cast from
the Indian non-mulberry tasar silkworm
Antheraea mylitta (which as mentioned pre-
viously, unlike Bombyx mori silk, contains a
natural RGD sequence) displayed a transpar-
ency (94.4 ± 0.006%) and a refractive index
(1.44 ± 0.03) suitable for corneal repair.[124]
These films supported the sprouting,
migration, attachment, and proliferation of
epithelial cells and keratocytes from rat cor-
neal explants to form complete cell sheets.
Further, the films supported the growth of
corneal limbal stem cells from the explants.
Silk films cast from Bombyx mori silk have
also been shown to support the adhesion and
growth of human corneal epithelial cells as
confluent epithelial sheets similar to those on
the amniotic membranes used clinically.[125]
Further, silk film topography[126] and biofunc-
tionalization[127] can be optimized to enhance
corneal epithelial cell interactions.
Following implantation into the cor-
neal pockets of rabbits, acellular silk films
made from Antheraea mylitta silk remained
transparent and showed no signs of neo-
vascularization.[124] The films remained
intact for at least two months and had no
adverse effect on tear production, intraocular
pressure, electrophysiology of the eye, or the
histology or ultrastructure of the cornea. Silk
film degradation can be controlled through
modification of its β-sheet content and this
has been extensively used to optimize silk
Figure 5. Implantable poly(acrylic acid)-silk fibroin microneedles for controllable vaccine
biomaterial properties, including those of release kinetics and enhanced immunogenicity. A) Schematic representation of the com-
silk films for corneal applications.[59,128] posite microneedle fabrication process, showing fabrication of the silk fibroin tips followed
Silk films are also extensively explored by poly(acrylic acid) (PAA) pedestals. B) Photograph of the composite microneedles. C) Con-
for engineering corneal stroma[12,129] and focal microscopy images of composite microneedles loaded with fluorescently labeled model
the development of in vitro corneal models proteins in the silk tip (blue) or PAA pedestals (red) and D) delivery of fluorescently labeled
incorporating epithelium, stroma, and model proteins to murine skin showing burst release and fast clearing from the PAA ped-
estal and slow, sustained release from the silk tip over 4 days (non-methanol-treated silk) or
innervation.[129b] In addition to corneal repair 16 days (methanol-treated silk). i.d. inj refers to the intradermal injection control. Adapted with
and regeneration, silk films are also being permission.[40a] Copyright 2014, Wiley-VCH.
investigated as a substrate for the develop-
ment of retinal prostheses. Recently reported
retinal prostheses consisting of semiconductive poly(3-hexylth- and laser micromachining.[40a] DeMuth et al. proposed an
iophene) and conductive poly(3,4-ethylene-dioxythiophene)- interesting approach to develop implantable-tip composite
poly(styrenesulfonate) layers spin-coated onto silk films were microneedles using poly(acrylic acid) (PAA) and Bombyx mori
shown to restore light sensitivity and visual acuity of the pri- silk fibroin for sustained vaccine delivery (Figure 5).[40a] Recent
mary visual cortex in a well-established rat model of retinitis studies have shown the importance of antigen and adjuvant
pigmentosa.[130] delivery kinetics in developing an optimal immune response,
Another promising application of silk films involves and persistent antigenic and inflammatory signals have been
the introduction of various topographical features, such as shown to elicit stronger responses when compared with tran-
microneedles. Microneedles are a minimally invasive and pain- sient bolus vaccine exposure.[97,133]
less alternative to hypodermic needles for drug administra- Silk has the advantage of controlled cargo release over time
tion. Silk microneedles have been made using micromolding and can protect temperature-sensitive cargoes at elevated tem-
approaches and a variety of masters, including those made by peratures, potentially allowing elimination of the “cold chain”
thermal drawing,[131] micromilling followed by wet etching,[132] that limits the availability of vaccines in developing countries.[134]
The composite microneedle consists of a silk fibroin tip loaded

with the vaccine of interest and vaccine-loaded PAA microneedle
pedestals.[40a] Upon exposure to the aqueous environment of the
skin, the PAA rapidly dissolves within hours, releasing a bolus
vaccine injection, while the silk fibroin tips remain implanted
in the skin, releasing the vaccine over days. The microneedles
were demonstrated to easily penetrate murine skin, with inser-
tion occurring several hundred micrometers below the skin
surface. The skin healed within a day of patch application. A
fluorescently labeled model protein was used to demonstrate
vaccine release, and rapid PAA-delivered cargo clearance was
observed from the treatment site within 24 h, followed by a slow
release of silk-encapsulated cargo at the treatment site between
4 days (when silk fibroin was not treated with methanol) and
>16 days (when silk fibroin was treated with methanol), indi-
cating the ability to tune the drug release. When used to deliver
a model whole-protein vaccine, the composite platform resulted
in over a tenfold increase in antigen-specific T-cell and humoral
immune responses when compared to traditional immuniza-
tion approaches. Notably, the microneedles were stored at room
temperature for two months prior to testing, demonstrating the
potential of this technology to eliminate the cold chain.
Figure 6. Silk-based devices for fracture fixation. A) scanning electron

9.3. Silk Scaffolds microscopy image of a silk fibroin screw. Scale bar is 1 mm. B) Silk fibroin
screw inserted into a rat femur at 4 weeks postsurgery. C,D) Cross-sec-
tions of the silk fibroin screw inserted into a rat femur at 4 weeks post-
3D porous silk scaffolds can be manufactured via a number surgery; sections stained with H&E and Masson’s trichrome, respectively.
of approaches,[42] including the use of sodium chloride as a Adapted with permission.[139] Copyright 2014, Macmillan Publishers.
porogen leached from aqueous or organic silk solutions or fol-
lowing freezing and lyophilization of aqueous silk solutions.[40c] to demonstrate the feasibility of the manufacturing and inser-
Early applications of these biomaterials predominantly focused tion process. The rats were mobile immediately following sur-
on bone replacement and regeneration[135] due to the high gery and showed no visible signs of pain. The screws were well
mechanical strength of silk scaffolds and the potential for tolerated for up to 8 weeks, with early signs of resorption and
further reinforcement using degummed silk fibers, which formation of new bone evident around the threads of the screw.
approached the mechanical properties required for load-bearing These devices are particularly appealing due to the potential
bones.[64] Silk scaffolds have since found applications in a range for malleability when hydrated, which will allow shaping of
of tissue engineering procedures for replacement and regenera- the fixation plates for unique anatomical locations during sur-
tion of tissues,[129b,40c] as well as in the development of 3D in gery, as well as providing the potential to incorporate BMP-2
vitro tissue models.[129b,40c] or antibiotics directly into the fixation devices to increase their
Recently, silk-based (Bombyx mori) fracture fixation devices functionality.
have been developed using an approach that differs from
the use of traditional highly porous silk scaffolds. Metal alloys
are the current gold standard for fracture-fixation devices, 9.4. Electrospun Silk Biomaterials
despite issues arising from the extreme mechanical mismatch
with native bone, which can have profound effects on wound Electrospinning has emerged as a popular technique for the
healing and long term viability of the devices, particularly in development of biomaterials due to the extracellular matrix
pediatric patients.[137] Resorbable fixation devices, such as those (ECM)-like fibrous nature of the nonwoven matrices and the
made of poly(lactic-co-glycolic acid) (PLGA), poly(glycolic acid) control over fiber properties that can be achieved by tuning the
(PGA), and, recently, silk, have the potential to address the dis- electrospinning parameters. Electrospun silk fibroin has been
advantages of metallic fixation devices. Silk, in particular, has explored for a range of applications, including wound dress-
the advantage of maintaining high strength while eliciting a ings,[114,140] bone[141] and ligament[142] replacements, and vas-
low inflammatory response. Furthermore, silk degradation does cular grafts.[143] Small diameter electrospun silk conduits have
not generate an acidic microclimate, as is typically observed for the potential to address the unmet need for off-the-shelf small
solid PLGA and PGA devices.[138] Bombyx mori silk fibroin fixa- diameter grafts with mechanical properties that match those of
tion devices, including bone plates and bone screws, were man- native vessels and support appropriate endothelial and blood
ufactured from lyophilized regenerated silk fibroin by casting cell interactions. In a comprehensive in vivo study of small
in hexafluoroisopropanol, followed by machining into desired diameter electrospun silk graft performance, Filipe et al. have
shapes (Figure 6).[139] Silk screws were successfully inserted demonstrated the production and performance of electrospun
into the hind limbs of rats (a nonfunctional preclinical model) acellular silk fibroin grafts generated under aqueous conditions
expressing cells and increases in collagen,

elastin, and proteoglycan production.[143c]
This study demonstrates the potential for the
use of silk fibroin in small diameter vascular
graft applications and the findings warrant
further preclinical testing in large animal
models.
9.5. Hydrogels
Silk fibroin hydrogels have emerged as

promising platforms for the delivery of
small molecular weight drugs, biologics,
and cells (reviewed in ref. [144]), as well as
for mimicking of the ECM in 3D in vitro
tissue models (reviewed in ref. [145]) and
in vivo tissue fillers.[146] A classification
of physically and chemically cross-linked
Bombyx mori silk hydrogel is useful. The
chemically cross-linked forms will not be dis-
cussed here (reviewed in ref. [144b]), because
these systems do not capitalize on the facile
self-assembly process of silk fibroin, a fea-
ture that sets it apart from many other (bio)
polymer-based hydrogels. This ability to self-
Figure 7. Electrospun silk fibroin biomaterials for vascular applications. A) Electrospun silk assemble arises because silk I can undergo a
fibroin graft morphology. Scale bar is 10 µm. B) Electrospun silk fibroin grafts implanted in a transition to silk II, which is rich in β-sheets,
rat model of abdominal aortic replacement: at the time of implantation (left), 6 weeks (middle),
by the exclusion of solvating water molecules
and 24 weeks (right) postimplantation. C) Endothelialization of implanted silk fibroin grafts
as demonstrated by vWF+ staining (red) at 6 weeks postimplantation in distal (left), middle from the hydrophobic domains of the silk
(middle), and proximal (right) regions. D,E) In vitro assessment of blood compatibility of block copolymer. The GY sequences are key
electrospun silk fibroin biomaterials, compared to ePTFE, as indicated by formation of a fluo- drivers for the formation of β-sheets, while
rescent fibrinogen network (D) and whole blood incubation (E). Adapted with permission.[143c] the exact molecular abundance and compo-
Copyright 2018, Elsevier. sition allows fine tuning of the solution–gel
transition process. Furthermore, pH respon-
in the absence of chemical cross-linkers (Figure 7).[143c] The sive elements within the silk structure allow silk to adopt a
electrospun silk fibroin grafts were significantly more elastic more ordered state (reviewed in ref. [147]).
when compared with the gold-standard synthetic polytetra- Many different triggers have been used to control the solu-
fluoroethylene (ePTFE) grafts (4.2 ± 0.5 MPa vs 31.9 ± 1.3 MPa) tion–gel transition, including but not limited to i) vortexing, ii)
and more closely matched the elasticity of native rat vessels ultrasound, iii) temperature, iv) osmotic stress, v) pH, vi) CO2
(2.1 ± 1.0 MPa), while demonstrating adequate burst pressure acidification, vii) nonsolvent induced phase separation, and viii)
(849 mmHg) and suture pull-out strength (0.86 N). In vitro, the direct electric current (reviewed in ref. [148]). Self-assembling
electrospun silk fibroin demonstrated excellent endothelial cell silk fibroin hydrogels show shear thinning, making them ideal
interactions and blood compatibility. for injection and minimally invasive procedures. Treatment
These characteristics are critical for the successful develop- protocols are now available that yield self-assembling hydro-
ment of next-generation vascular grafts, because only grafts that gels in the absence of organic solvents, chemical cross-linkers,
have both outstanding hemocompatibility and support endothe- or UV irradiation. However, a potential disadvantage of self-
lialization will yield long-term performance. In an in vivo rat assembling silk fibroin hydrogels is that physically cross-linked
model of abdominal aortic replacement, electrospun silk fibroin systems (i.e., those with high β-sheet content) are opaque, due
scaffolds demonstrated excellent surgical handling and patency to the formation of nanocrystallites, and they are brittle, as they
for up to six months and outperformed the gold-standard cannot undergo long range displacements, resulting in low
ePTFE grafts. Silk fibroin grafts supported rapid endotheliali- elastic behavior and plastic deformation at strains >10%.
zation, with endothelial cells present as early as 3 weeks post- Electric fields yield silk fibroin hydrogels (also known as
implantation and an almost complete monolayer forming by e-gels) that differ from most other physically cross-linked
6 weeks (Figure 7). By contrast, the ePTFE grafts remained types.[149] First, the secondary silk structure of e-gels is domi-
largely uncovered by endothelial cells even at 24 weeks postim- nated by helical conformations, whereas other (pH-induced)
plantation. The silk fibroin grafts showed intimal hyperplasia hydrogels are rich in β-sheets. Second, e-gel formation is
stabilization by 6 weeks, with smooth muscle cells (SMCs) reversible: a switch in the DC current induces a migration of
showing a phenotypic switch to the less proliferative SMC-α silk to the new positive electrode (this is possible because of
the absence of strong β-sheets). Third, e-gels have strong adhe- of trophic and angiogenic factors. Furthermore, transplantation
sive properties that are typically absent from other silk fibroin of minimal islet cell grafts and MSCs (serving as a supporting
hydrogels. Fourth, e-gels have outstanding elastic properties stromal cells) using self-assembling silk fibroin hydrogels
and can withstand strains up to 2500%.[149] resulted in euglycemia control in 75% of the transplanted mice
The assembly of silk micelles into an e-gel at the electrode at day 37, whereas no other treatment combinations success-
is completed within minutes. The local drop in pH at the posi- fully abolished diabetes.[154] The use of a silk fibroin hydrogel
tive electrode below the isoelectric point of silk screens repul- maximized the clinical performance; however, histological
sive charges and enables hydrogel formation.[150] Overall, e-gels examination showed an unintended complication as the MSCs
have been proposed to support a range of biomedical applica- differentiated into osteoblasts at day 42. MSCs are responsive
tions (e.g., adhesives for medical devices, sensors, etc.) and to environmental cues (e.g., mechanical forces, cytokines, etc.),
proof of principle studies are eagerly awaited. which indicates that (silk) hydrogels still require fine-tuning to
Self-assembling Bombyx mori silk fibroin hydrogels with a ensure the final desired outcome.
high β-sheet content have also been explored for a broad range
of biomedical applications. For example, self-assembling silk
fibroin hydrogels were loaded with doxorubicin and adminis- 9.6. Particles
tered locally to breast tumors in mice. Locally administered silk
fibroin hydrogels loaded with doxorubicin provided a signifi- The preclinical development of silk (nano)particles is often
cant reduction in primary tumor growth and metastasis when aimed at the delivery of cytotoxic small-molecular-weight anti-
compared to equivalent doses of doxorubicin administered sys- cancer drugs (reviewed by ref. [70]). Entrapment into a (nano)
temically.[151] A parallel study that used the same animal model, particle changes the pharmacokinetic and biodistribution char-
drug, dosing schedule, and treatment strategy showed that the acteristics of the payload, as these characteristics are now dic-
heparin-modified polyethylene glycol (PEG) hydrogels were tated by the carrier and not the physiochemical properties of the
outperformed by the self-assembling silk hydrogel.[152] Self- drug. Therefore, engineering the carriers opens up new possi-
assembling silk fibroin hydrogels that use sonication as a trigger bilities for tuning the overall drug performance; for example,
are ideal for the delivery of biologics. The payload is added fol- by altering the residence time in the blood or the uptake
lowing sonication and during the solution-gel transition time mechanism into target cells. Nanoparticles are often proposed
window. For example, self-assembling silk fibroin hydrogels for solid tumor targeting, as they can exploit the leaky vascula-
have been examined for the delivery of bevacizumab (a mono- ture and reduced lymphatic drainage associated with tumors,
clonal antibody of antivascular endothelial growth factor) for which results in enhanced permeability and retention (EPR) at
the treatment of age-related macular degeneration.[153] the target tissue.[155] Inclusion of a targeting ligand can further
Direct comparison of a silk fibroin hydrogel formulation with increase the specificity. The EPR effect exploits pathophysi-
the current standard treatment showed that intravitreal injec- ology, but full clinical exploitation remains to be realized.[156]
tion in healthy rabbits significantly improved the drug levels Nanoparticles designed for EPR-mediated tumor targeting are
in the vitreous and aqueous humor when compared to the typically injected into the blood circulation and must therefore
commercial liquid formulation. The bioavailability of the com- be compatible with blood. Silk fibroin nanoparticles showed
mercial product and the silk fibroin hydrogel formulation was very low plasmatic coagulation and the observed response was
similar, but the terminal half-life for the silk hydrogel was two- significantly better when compared to silica nanoparticles.
fold to threefold higher. This improvement would be expected Furthermore, under simulated venous blood flow, the silk and
to reduce the number of intravitreal injections required.[153] PEGylated silk fibroin nanoparticles also showed low inflam-
Although the results are encouraging, further optimization of mation when compared to silica nanoparticles.[72] Overall, these
the silk fibroin hydrogel is warranted to reduce the initial burst initial studies on the hemocompatibility of silk fibroin nanopar-
release of bevacizumab and the placement of the silk hydrogel, ticles are encouraging.
because vitreous administration has the potential to obstruct Early studies set out to explore techniques for the manu-
the light path into the eye and thereby limit vision. facturing of silk fibroin nanoparticles and to establish their
Self-assembling silk hydrogels are emerging as useful tools respective loading capabilities using (model) drugs (reviewed
for the therapeutic delivery of (stem) cells. For example, pan- by ref. [157]). Two of the early manufacturing techniques used
creatic islet transplantation is plagued by a functional decline to generate silk fibroin nanoparticles were nanoprecipitation[158]
and decreased viability of the islets during the peritransplanta- and capillary microdot printing.[159] Capillary-dot microprinted
tion period, so self-assembling silk fibroin hydrogels have been nanoparticles loaded with curcumin showed extended release
examined as a potential delivery system.[154] Silk self-assembly profiles and a higher in vitro efficiency against breast cancer
was initiated by vortexing, the islet cells were added prior to the cells when compared to silk/chitosan composite nanoparti-
completion of the solution–gel transition, and the mixture was cles.[159] Nanoparticles generated from Bombyx mori and Anthe-
injected into the epididymal fat pad of diabetic mice. Functional raea mylitta (the tropical tasar silkworm) silks were stable,
tests showed that silk fibroin hydrogels loaded with pancreatic spherical, negatively charged, and 150–170 nm in diameter, and
islets were able to control glucose levels within 4 days, whereas they showed no cytotoxicity at the tested concentrations.[160]
this time was extended to 14 days in the absence of the silk car- The delivery of small molecular weight drugs with (silk)
rier matrix. Intraperitoneal glucose tolerance tests showed that nanoparticles changes their uptake mechanisms (and their
cotransplantation of mesenchymal stem cells (MSCs) with the susceptibility to drug efflux pumps) from passive diffusion
islets improved the function of the graft through the production across the plasma membrane to an energy-dependent endocytic
uptake (independent of drug efflux pumps). For these reasons, self-assembly of SELP nanofibers,[176] while macroscopic silk
silk fibroin nanoparticles were able to improve anticancer drug constructs have been patterned using ion beam lithography
delivery into drug resistance breast cancer cells.[161] to yield diverse silk-like constructs with defined shapes at the
Once inside the cells, the payloads on silk fibroin nanoparti- nanometer scale.[177] Other examples include engineered silk
cles can also be activated within lysosomes (i.e., lysosomotropic oligonucleotide conjugates that direct silk assembly into a par-
drug delivery) by the low pH and the proteolytic enzymes of lys- allel, antiparallel, and branched configurations (reviewed in
osomes, given the correct intracellular trafficking of the nano- ref. [178]). The timely review by Aigner et al. provides extensive
particles.[162] The lysosomal environment not only triggers drug insight into this important branch of silk research.[167a] Here,
release but is also the site of silk fibroin nanoparticle degrada- we only provide a selection of a few examples of recombinant
tion.[51] Silk fibroin (nanoparticles) can stabilize a broad spectrum silks (Table 4), with a specific reference to silk (nano)particles.
of payloads by tailoring the water content, locking the payload into Recombinant silk proteins have been inspired by Araneus
place, buffering the microenvironment, and restricting the access diadematus fibroin 4 (ADF4); ADF4 is from the common Euro-
of degradative enzymes (reviewed in ref. [134]). For example, pean garden spider. ADF4 is one of the most widely studied
entrapment of L-asparaginase into Bombyx mori silk fibroin nan- spidroins[5a] and has been used to prepare microcapsules for
oparticles resulted in an increased resistance to enzymatic deg- drug delivery by exploiting its self-assembly at an emulsion
radation, better stability in serum, prolonged storage stability in interface.[187] The resulting spider silk-based carriers are useful
solution, and minimal leakage of the enzyme from the carrier.[163] for encapsulating low molecular weight drugs under mild con-
PEGylation of silk fibroin nanoparticles has been exploited to ditions, which maintains the activity of the payload.[187] Expo-
improve colloidal stability and to tailor drug release and carrier sure of ADF4-like silks to potassium chloride (>400 × 10−6 m)
degradation.[164] Magnetically responsive, drug-loaded silk fibroin generates particles,[188] which have been extensively character-
nanoparticles have also been developed by seeding silk with ized for their ability to entrap and release (small molecular
Fe3O4 nanoparticles.[165] Subjecting these silk nanoparticles to a weight) payloads.[189] Silk sequence modifications are now
magnetic field in the tumor area allowed their enrichment in the yielding cationic proteins that can be loaded with low and high
tumor, thereby promoting drug accumulation and an improved molecular anionic payloads (e.g., nucleic acids).[190] Libraries of
antitumor response.[165] Silk fibroin nanoparticles have also been silk proteins containing modifications, such as the RGD inte-
combined with other silk formats (for example, silk fibroin hydro- grin binding domain and the Tat cell penetrating peptide, have
gels) to yield first generation all-silk dual-drug delivery systems.[166] been designed to enhance cellular uptake and drug delivery.[191]
The first inroads have been made to unravel the mechanisms
for endocytic (e.g., caveolae, clathrin-mediated) uptake of nano-
10. Bioengineered Silks particles into cells using putative chemical inhibitors.[192]
Recombinant technologies have also been exploited to gen-
Recent efforts have focused on developing recombinant forms erate silks inspired by major ampullate spidroin 1 (MaSp1)
of silks that can be altered at the sequence level to achieve spe- from the Gold Orb-web spider (Nephila clavipes). For example,
cific modalities, for example, for biomedical use.[167] Recom- MaSp1-like proteins have been modified with poly(l-lysine)
binant approaches are unique because they allow the design cationic sequences to allow complexation with nucleic acids via
and manufacture of bespoke “silks.”[168] For example, key ele- electrostatic interactions for use in gene delivery. These engi-
ments of the spider silk sequence can be lifted and combined neered protein vectors demonstrated excellent DNase resistance
with polylysine to develop novel biopolymers for the delivery and gave transfection efficiencies similar to those achieved with
of genetic material (reviewed in ref. [169]). Chimeric proteins, the commercial reagent Lipofectamine 2000. The silk sequence
such as silk-collagen-like proteins[170] or silk-elastin-like pro- has also been fused with tumor-homing peptides (e.g., F3,
teins (SELPs), are exciting new materials (although the SELPs Lyp1 CGKRK) and attachment ligands (e.g., RGD to exploit cell
have been extensively studied over the past three decades; binding and receptor-mediated endocytosis) to enhance their
reviewed in ref. [171]). SELPs are facile biopolymers that can be targeting capabilities.[61c,93,166] These recombinant silks formed
fine-tuned to achieve a desired form and function; for example, nanometer-sized globular complexes with plasmid DNA (150–
SELP micelles have been designed for anticancer drug delivery 250 nm in diameter), and they demonstrated significantly
that exploits both passive and active tumor targeting. SELPs have improved target specificity for melanoma and highly metastatic
also been used to generate hydrogels intended for local drug human breast cancer cells.[93,166] A silk vector harboring the
release.[171] Many other studies have examined the performance tumor-homing peptide F3 showed minimal toxicity in healthy
of “biopolymer alloys” by blending silk with another biopolymer, cells, the best tumor specificity, and a capability to deliver its
such as tropoelastin,[172] collagen,[173] and fibronectin[174] or payload in a human orthotopic breast cancer model.[193] Overall,
have included inorganic ceramics (reviewed in ref. [175]) to these studies demonstrate the potential of silk-based delivery
generate new material systems with expanded function. One of systems as nonviral gene delivery vectors.
the hallmarks of silk is its inherent ability to organize structures
at the nanometer scale; these structures then assemble, grow,
and ultimately produce macroscale constructs with defined 11. (Old) New Silk Industries: Opportunities
function. The ability of silk to work seamlessly across several
and Challenges for the Road Ahead
orders of magnitude is exciting and has motivated the devel-
opment of engineered nanoscale systems. For example, atomic Current production routes for Bombyx mori silk are well estab-
force microscopy has been used to drive and direct the lished and are the foundation of several products in routine
Table 4. Examples of silk and functional motifs used for engineered silk biomaterials.
Motif Amino acid sequence Function Reference

Major ampullate spidroin 1 (MaSp1) GRGGLGGQGAGAAAAAGGAGQGGYGGLGSQG Repeated domain [179]
from Nephila clavipes
GAGAAAAAGGAG Hydrophobic block [180]
QGGYGGLGSQGSGRGGLGGQ Hydrophilic block [180]
ADF-4 from the garden spider GSSAAAAAAAASGPGGYGPENQGPSGPGGYGPGGP Repeated domain [181]
Araneus diadematus
B. mori silkworm heavy chain GAGAGS Beta-sheet motif [182]
[GERGDLGPQGIAGQRGVV(GER)3GAS]8GPPGPCCGGG [TGRGDSPAS]8 Repeated domain [183]
Dentin matrix protein 1 (CDMP1) RGDNPDNTSQTGDQRDSESSEEDRLNTFSSSESQSTEEQGDSESNESLSL- Controlled hydroxyapatite growth [39a]
SEESQESAQDEDSSSQEGLQSQSASRESRSQESQSEEDSRSEENRDS-
DSQDSSRSKEESNSTGSTSSSEEDNHPKNIEADNRKLIVDAYHNKPIG-
DQDDNDCQDGYLE
R5 unit of silaffin-1 precursor SSKKSGSYSGSKGSKRRIL Induce silica precipitation [184]
polypeptide from Clavulinopsis fusiformis
Bone sialoprotein MKTALILLSI LGMACAFSMK NLHRRVKIED SEENGVFKYR PRYY- Induce bone regeneration [93]
LYKHAY FYPHLKRFPV QGSSDSSEEN GDDSSEEEEE EEETSNEGEN
NEESNEDEDS EAENTTLSAT TLGYGEDATP GTGYTGLAAI QLPKKAGDIT
NKATKEKESD EEEEEEEEGN ENEESEAEVD ENEQGINGTS TNSTE-
AENGN GSSGGDNGEE GEEESVTGAN AEDTTETGRQ GKGTSKTTTS
PNGGFEPTTP PQVYRTTSPP FGKTTTVEYE GEYEYTGANE YDNGYEIYES
ENGEPRGDNY RAYEDEYSYF KGQGYDGYDG QNYYHHQ
Cell penetrating peptide ppTG1 GLFKALLKLLKSLWKLLLKA Delivery system into cells [61c,185]
Tumor homing peptide F3 KDEPQRRSARLSAKPAPPKPEPKPKKAPAKK Target specificity to tumor cells [93,186]
RGD RGD Cell adhesive and integrin mediated [61c,184a]
delivery
clinical use today. Following a period of decline, global silk pro- This, in turn, is likely to simplify the regulatory requirements
duction is growing once again and has the capacity to keep up that currently mandate a well-defined material stock (although
with a growing silk demand for (bio)medical use.[19] However, these regulations vary depending on the specific classification
sericulture is an agricultural process that depends on several fac- of the final product as “medical device,” “excipient,” “advanced
tors, including, but not limited to, climate, seasonal variations, therapy medicinal product,” “novel chemical entity,” etc.).
disease and pest control, and the susceptibility of silkworms to Full reverse engineering of the silk cocoon is complex from
common pesticides used in other agriculture sectors (within both industrial and regulatory perspectives. At present, all
geographical proximity, with fatal consequences for the silk- licensed (Bombyx mori) silk products in the USA and EU are
worm). The silk community should make a concentrated effort considered medical devices based on the nascent, but deg-
to work with well-controlled silk cocoon stocks with a known ummed, silk fiber (Tables 1, and 2). By contrast, reverse engi-
process history in order to propel the silk research community neered silk is marketed as cosmetics to navigate regulatory
into working practices that align with Good Laboratory Practice. challenges. Irrespective of the medical or cosmetic use, the cur-
This type of approach would be invaluable for improving com- rent silk production technologies are batch processes, which go
parisons across different studies and accelerating clinical trans- against the current industrial efforts aimed at continuous manu-
lation. While public guidelines exist for CE-marked organic facturing to ultimately improve product reliability and cut costs.
sericulture, the production of Bombyx mori silk for medical use Manipulating (the germ line of) silkworms by genetic engi-
is shrouded in secrecy and proprietary protocols. For example, neering opens up new possibilities to generate improved silks,
the domesticated Bombyx mori silkworm line used for silk for example, by inserting spider silk sequences into Bombyx
sutures and surgical meshes is a well-kept secret. mori silk to improve the mechanical properties of the in vivo
The optimal environmental condition during cocoon spin- spun fiber.[195] Another possibility is to exploit the silkworm as
ning is another unknown. Laboratory experiments indicate that a biosynthetic host to synthesize xenogenic proteins or func-
silk cocoons spun in a low humidity environment do not require tional silks (e.g., insertion of green fluorescent protein, RGD
degumming by boiling in an aqueous alkaline solution; instead, sequences, etc.[196]). Improvements in genome editing are
physical manipulation is sufficient for quantitative removal of likely to increase the protein yields, ultimately leading to new
sericin.[194] Of course, adequate silk cleaning is critical for pro- silkworm lines (e.g., that produce modified silk sequences, etc.)
ducing hypoallergenic silk. Eliminating the boiling step also of appreciable economic value.
preserves the silk structure, thereby yielding a monodispersed Recombinant proteins are routinely used in the clinic and are
biopolymer (rather than polydispersed silk fibroin fragments). manufactured on a large scale by the pharmaceutical industry.
Table 5. Examples of current companies with R&D and/or silk-based products for biomedical applications.
Company Location Silk Products Clinical trial(s)

Sofregen Inc. Medford, MA, USA Bombyx mori SERI Surgical Scaffolda),b) NCT02033590
Silk fillersc) NCT01389232
NCT01914653
NCT01981044
NCT02293798
NCT02016612
Vaxess Inc. Boston, MA, USA Bombyx mori Encapsulating and stabilizing payloadsc)
Silk Therapeutics Inc. Medford, MA, USA Bombyx mori Anti-aging skin carea)
Cocoon Biotech Inc. Cambridge, MA, USA Bombyx mori Silk-based drug deliveryc) (e.g., hydrogels, micro-
spheres for ophthalmic and osteoarthritis)
Silk Technologies Ltd. Plymouth, MN, USA Bombyx mori Topical occular therapy (e.g., dry eye); silk-derived
protein pharmacological active ingredientc)
Oxford Biomaterials Ltd. Oxford, UK Bombyx mori and others Vascular graftsc)
Orthox Ltd. Oxford, UK Bombyx mori FibroFix Meniscusb) NCT02732873
Spidrex tissue scaffoldsc) NCT02205645
Neurotex Ltd. Oxford, UK ND Spidrex nerve conduitc)
AL.PRE.TEC. S.r.l. San Donà di Piave, Italy Bombyx mori Textiles for skin diseases (e.g. dermatitis)a) andc),
DermaSilk (see Table2)a)
Suzhou Soho Biomaterial Science Suzhou, Jiangsu Prov- Bombyx mori Sidaiyi wound dressinga) biomedical use of other silk NCT01993030
and Technology Co., Ltd, China ince, China formatsc)
Zhejiang Xingyue Biotechnology Hangzhou, China Bombyx mori HQ Matrix medical wound dressingb); HQ Matrix NCT01993030
Co., Ltd. Soft Tissue Mesh (hernia repair)b) NCT02487628
Daewoong-Bio Inc. Seoul, South Korea Bombyx mori Tympasil silk patcha) (tympanic membrane Yes, but not registered at
perforation) ClinicalTrials.gov
AMSilk GmbH Planegg, Germany Recombinant spider silk Cosmetic (silk beads, silk gels sold to cosmetic Yes, but not registered at
industry)a) ClinicalTrials.gov
Biosteel fibersc)
Surgical meshesc)
Coating of breast implantsb)
Spiber Technologies AB Stockholm, Sweden Recombinant spider silk Spiber genetically modified for new biomedical
functionsc)
SolvNT protein solubility tagc)
Spiber Inc. Yamagata, Japan Recombinant spider silk QMONOS silk fibers, apparel and automotivec),
biomedical secondary
Bolt Threads Inc. Emeryville, CA, USA Recombinant spider silk Silk fiber and Bolt Microsilk clothing (c) and lim-
iteda)). Biomedical use undisclosed
The Synthetic Bioproducts Center Logan, UT, USA Recombinant spider silk Proprietary recombinant silks for biomedical
applicationsc)
Kraig Biocraft Laboratories Inc. Ann Arbor, MI, USA Bombyx mori Silk modified with spider silk sequences & produced
in silkworms. Dragon Silk and Monster
Silk lead products as
ballistic shoot packs.c) Biomedical use
undisclosed
a)Commercial
product/marketed; b)In clinical trials; c)In development.
Therefore, the necessary expertise exists within the health- constructs, the very high glycine content of the protein, and
care sector to manufacture these complex products to clinical the high molecular weight of the product (250–320 kDa). These
specifications. Already, today, recombinant DNA technology, challenges can be addressed by using genetically modified
in combination with Escherichia coli expression systems, has E. coli with an elevated glycyl-tRNA pool, which give native-like
revolutionized silk research[167a] and is opening up new com- silk proteins, albeit with a low protein yield.[198] Another com-
mercial avenues (Table 5). However, silk presents a number of plication is that silk inspired proteins with repeated domains
specific challenges; for example, the silk molecules cannot be containing Ala-rich sequences are poorly soluble in water and
post-translationally modified, but this is required to faithfully buffers. Inclusion of a cationic histidine tag with the hydro-
mimic the silk protein.[197] The expression of native-sized silk phobic crystallizable silk sequences improves the aqueous
proteins is also not possible using standard E. coli expression solubility of the resulting protein.[61c] The economic issues
systems because of the highly repetitive nature of the gene and greenhouse gas emissions associated with E. coli cultures
are other critical aspects that must be considered for the currently working with recombinant silks are generating fibers.
development of a sustainable recombinant silk industry.[199] The remarkable mechanical properties of spider silk, coupled
with its processing under ambient conditions in an all aqueous
environment, might serve as an inspiration for this new industry.
11.1. Staying on the “Silk Road” The Synthetic Bioproducts Center, under the leadership of
Randy Lewis (Utah State University, Logan, UT, USA), produces
Perhaps one of the most important aspects to consider synthetic spider silks for biomedical engineering applications
regarding the future of the biomedical use of silk is the using a range of expression hosts, including genetically modified
safeguarding of the silk development pipeline. Silk is a truly goats. These genetically engineered dairy goats carry the dragline
amazing biopolymer that has inspired generations of scientists silk genes of the Nephila clavipes (identified by the Lewis lab[202])
and is likely to continue to do so. However, researchers must in mammary gland cells and excrete soluble silk in their milk.
not get carried away or overpromise. In a globally connected This silk can be extracted and subsequently spun into fibers. The
world, with many clinically unmet needs, the news of promising genetically modified goats were developed by Nexia Biotechnolo-
biomedical research has the potential to make headlines simply gies Inc. (Quebec, Canada) in the early 2000s and subsequently
by the fact that the material is “silk.” We must not exploit the acquired by Randy Lewis. Nexia Biotechnologies and the Materials
familiarity of the general population with common silk for the Science Team of the U.S. Army and Soldier Biological Chemical
purpose of short term gains. The road to the clinical translation Command (Natick, MA, USA) expressed the silk proteins ADF-3/
of basic biomedical research is a long and tortuous one—this MaSp2 and MaSp1 for the first time in mammalian cells,[203]
also applies to silk research. The path is difficult and requires thereby laying the foundation for subsequent work in goats.
a carefully measured balance of optimism (to inspire people) AMSilk GmbH (Planegg/Munich, Germany) is a spin-out
and realism (to avoid a silk bubble). It is important to learn company based on seminal work by Scheibel and co-workers and
from past failures as well as from the challenges experienced is the world’s first industrial supplier of recombinant silk biopoly
in allied fields.[200] Clearly, silk has its limitations, as detailed mers across a range of applications. AMSilk exploits ADF4-
in this review; for example, i) it is an expensive biopolymer inspired silks (and others) to generate high quality materials for
when compared to mass-produced fully synthetic polymers, ii) apparel, cosmetic, industrial, and structural applications. Biosteel
sericulture is a labor-intensive agricultural process and highly fiber is their leading product, and it is at various developmental
responsive to its environment, which ultimately impacts silk stages for application in the footwear and automotive industries.
quality, and iii) the durability of recombinant spider silks might AMSilk received ISO 13 485 accreditation in 2017—a prerequi-
be challenging based on the simple premise that the material site for initiating clinical studies in humans. In February 2018,
has evolved in nature as a short-term high-performance mate- AMSilk announced the launch of the POSIS Phase I clinical trial,
rial (e.g., orb-weaver spiders repair or build a new web often in partnership with POLYTECH Health & Aesthetics GmbH.
daily). It is therefore important to consider alternative materials The trial examines the performance of silicone breast implants
as well that are able to perform, and perhaps surpass, the func- coated with ADF4-based silk (BioShield-S1) (SILKline) in several
tion of silk. The emergence of fully synthetic sutures is a testa- human subjects at a number of University hospitals in Austria
ment to this type of development. However, silk fibroin sutures (with the option to expand to other centers). This trial has been
still remain the first choice for specialized surgeries, indicating underpinned by both proprietary product development efforts
that silk should serve as a blueprint for next generation sutures. as well as animal studies available in the public domain. For
example, ADF4 coated silicone implants showed no acute toxicity
or immunogenicity and they reduced postoperative inflamma-
11.2. The Current Silk Drivers tion and minimized implant-induced capsule thickness and con-
traction when compared to uncoated control implants.[204]
Silk continues to inspire and serves as the thread for exploration Founded in 2008, Spiber Technologies AB (Stockholm,
new avenues: curiosity-driven research and learning from nature Sweden) produces silk biomaterials, which can be processed
are key elements for innovation. Newly emerging silk industries into a range of material formats (e.g., fibers, films, foams, coat-
are now translating research findings that go beyond fiber tech- ings, etc.). Their work is underpinned by academia–industry
nology[201] (Table 5). As these technologies, which often have their collaborations that support the preclinical research and devel-
origins in the academic setting, move from the public domain into opment pipeline exploiting spider silk for biomedical applica-
the industrial space, tracking their progress becomes more dif- tions (e.g., cell–matrix interactions, culture matrices, surface
ficult. However, the first products have entered clinical trials and modifications, electrochemical meshes, etc.).[205] Scientists
products are emerging on the market (Table 5 and detailed below). affiliated with Spiber Technologies AB have also unraveled the
Recombinant (spider) silks are the lifelines of many small to molecular mechanisms of spider silk spinning[206] in order to
medium sized enterprises. Spider silk is a prime focus, because develop better biomimetic fibers.[207]
this remarkable biopolymer cannot be obtained at an industrial The pH-sensitive N-terminal of silk enables the silk protein
scale by “farming.” Instead, genetic engineering and recom- to stay in solution at very high protein concentrations; thus,
binant expression systems are essential, and they also provide recombinant fusion of the N-terminal domain converted an
greater flexibility and rapid production of novel silk-inspired pro- aggregation-prone therapeutic protein to its hypersoluble coun-
teins. The (pharmaceutical) industry is accustomed to working terpart (this technology is now marketed as SolvNT).[208] This
with recombinant proteins, and these provide many opportuni- strategy also simplified protein purification, improved yields,
ties for patterning and proprietary knowledge. Many companies and allowed the expression of nontransmembrane proteins that
are otherwise refractory to recombinant production. SolvNT [1] a) J. Sparkes, C. Holland, Nat. Commun. 2017, 8, 594; b) D. Porter,
exemplifies how silk continues to amaze us, and goes beyond J. Guan, F. Vollrath, Adv. Mater. 2013, 25, 1275; c) D. Porter,
applications we are accustomed to. F. Vollrath, Z. Shao, Eur. Phys. J. E: Soft Matter Biol. Phys. 2005,
Spiber Inc. (Yamagata, Japan), founded by Kazuhide Seki- 16, 199; d) N. G. Rim, E. G. Roberts, D. Ebrahimi, N. Dinjaski,
M. M. Jacobsen, Z. Martin-Moldes, M. J. Buehler, D. L. Kaplan,
yama in 2007 as a start-up at Keio University, produces var-
J. Y. Wong, ACS Biomater. Sci. Eng. 2017, 3, 1542; e) D. Ebrahimi,
ious types of recombinant proteins, especially spider silk-like O. Tokareva, N. G. Rim, J. Y. Wong, D. L. Kaplan, M. J. Buehler,
proteins (note that Spiber Inc. is an independent company ACS Biomater. Sci. Eng. 2015, 1, 864.
from Spiber Technologies AB). The synthetic spider thread [2] a) C. Craig, Annu. Rev. Entomol. 1997, 42, 231; b) C. L. Craig, Spider-
QMONOS (based on the Japanese word kumonosu for spider webs and Silks: Tracing Evolution from Molecules to Genes to Pheno-
web) is a technology fiber that has been fed into several proof types, Oxford University Press, New York 2003; c) T. D. Sutherland,
of concept items, such as a child’s dress, and the North Face J. H. Young, S. Weisman, C. Y. Hayashi, D. J. Merritt, Annu.
branded MOON PARKA. The current target markets are the Rev. Entomol. 2010,55, 171; d) Q. Xia, S. Li, Q. Feng, Annu. Rev.
apparel and automotive segments, with healthcare materials Entomol. 2014, 59, 513; e) P. L. Babb, N. F. Lahens, S. M. Correa-
serving as an emerging future area. Garhwal, D. N. Nicholson, E. J. Kim, J. B. Hogenesch, M. Kuntner,
L. Higgins, C. Y. Hayashi, I. Agnarsson, B. F. Voight, Nat. Genet.
Another contender in the recombinant silk market space is Bolt
2017, 49, 895; f) N. Yonemura, F. Sehnal, J. Mol. Evol. 2006,
Threads Inc. (Emeryville, CA, USA), which produces silk fibers by 63, 42.
wet spinning (Engineered Silk) using advanced yeast expression [3] a) P. R. Laity, S. E. Gilks, C. Holland, Polymer 2015, 67,
systems. The recombinant silk is produced on a scale that makes 28; b) P. Laity, C. Holland, Int. J. Mol. Sci. 2016, 17, 1812;
it a viable contender for broad applications. To date, Bolt Threads c) M. Andersson, L. Holm, Y. Ridderstråle, J. Johansson, A. Rising,
has already introduced several engineered Silk fiber and Bolt Biomacromolecules 2013, 14, 2945; d) A. Rising, J. Johansson, Nat.
Microsilk products (e.g., ties, clothing, etc). Whether these mate- Chem. Biol. 2015, 11, 309.
rials will also enter the healthcare arena is not known at present. [4] a) B. Mortimer, C. Holland, F. Vollrath, Biomacromolecules
2013, 14, 3653; b) B. Mortimer, S. D. Gordon, C. Holland,
C. R. Siviour, F. Vollrath, J. F. C. Windmill, Adv. Mater. 2014, 26,
5179; c) G. R. Plaza, P. Corsini, E. Marsano, J. Perez-Rigueiro,
12. Conclusions M. Elices, C. Riekel, C. Vendrely, G. V. Guinea, J. Polym. Sci.,
Part B: Polym. Phys. 2012, 50, 455; d) G. B. Perea, C. Solanas,
In this progress report, we have unraveled some of the mys- N. Marí-Buyé, R. Madurga, F. Agulló-Rueda, A. Muinelo, C. Riekel,
teries of silk and critically examined the current and emerging M. Burghammer, I. Jorge, J. Vázquez, G. R. Plaza, A. L. Torres,
clinical uses of silk. We eagerly await further clinical reports, F. del Pozo, G. V. Guinea, M. Elices, J. L. Cenis, J. Pérez-Rigueiro,
especially on engineered second-generation silk materials, such Eur. Polym. J. 2016, 78, 129.
as “silk”-coated implants. For the past 5000 years, silk has capti- [5] a) T. Y. Lin, H. Masunaga, R. Sato, A. D. Malay, K. Toyooka,
vated humans, and it continues to amaze us as we explore new T. Hikima, K. Numata, Biomacromolecules 2017, 18, 1350;
applications. The silk biopolymer represents a pioneer material b) W. Abdel-Naby, B. Cole, A. Liu, J. Liu, P. Wan, R. Schreiner,
for medical applications today, and yet, even after many centu- D. W. Infanger, N. B. Paulson, B. D. Lawrence, M. I. Rosenblatt,
PLoS One 2017, 12, e0188154.
ries, it continues to be a valued suture material. As we continue
[6] a) A. D. Malay, R. Sato, K. Yazawa, H. Watanabe, N. Ifuku,
to extract secrets from silk, we will be able to develop new “old H. Masunaga, T. Hikima, J. Guan, B. B. Mandal, S. Damrongsakkul,
materials” to address current and future biomedical needs. K. Numata, Sci. Rep. 2016, 6, 27573; b) M. Humenik, T. Scheibel,
A. Smith, Prog. Mol. Biol. Transl. Sci. 2011, 103, 131; c) J. Guan,
Y. Wang, B. Mortimer, C. Holland, Z. Shao, D. Porter, F. Vollrath,
Acknowledgements Soft Matter 2016, 12, 5926.
[7] a) B. Madsen, Z. Z. Shao, F. Vollrath, Int. J. Biol. Macromol.
F.P.S. acknowledges funding support from EPSRC (EP/N03127X/1). 1999, 24, 301; b) Z. Z. Shao, F. Vollrath, Nature 2002, 418, 741;
J.R.K. acknowledges funding support from the Australian Research c) J. Perez-Rigueiro, M. Elices, G. Plaza, J. I. Real, G. V. Guinea,
Council (Grant # DP150104242). C.H. acknowledges funding support J. Exp. Biol. 2005, 208, 2633.
from EPSRC (EP/K005693/1). [8] A. A. Walker, C. Holland, T. D. Sutherland, Proc. Biol. Sci. 2015,
282, 20150259.
[9] C. A. Holland, A. E. Terry, D. Porter, F. Vollrath, Nat. Mater. 2006,
5, 870.
Conflict of Interest [10] a) M. A. Garrido, M. Elices, C. Viney, J. Perez-Rigueiro, Polymer
F.P.S. is supervising an MRes candidate who is sponsored by Allergan Inc. 2002, 43, 4495; b) T. A. Blackledge, R. A. Cardullo, C. Y. Hayashi,
K.N. has a collaborative project with Spiber Inc. (Yamagata, Japan). Invert. Biol. 2005, 124, 165; c) H.-P. Zhao, X.-Q. Feng, H.-J. Shi,
Mater. Sci. Eng., C 2007, 27, 675; d) C. Viney, in Conference Pro-
ceedings of the Society for Experimental Mechanics Series, Vol. 5,
Springer, New York 2013, p. 127.
Keywords [11] J. F. V. Vincent, O. A. Bogatyreva, N. R. Bogatyrev, A. Bowyer,
A.-K. Pahl, J. R. Soc., Interface 2006, 3, 471.
clinical uses, fibroin, recombinant silk, silk, spider silk [12] E. S. Gil, B. B. Mandal, S. H. Park, J. K. Marchant, F. G. Omenetto,
D. L. Kaplan, Biomaterials 2010, 31, 8953.
Received: April 29, 2018 [13] E. Morgan, Gossamer Days: Spiders, Humans and Their Threads,
Revised: August 4, 2018 Strange Attractor, London, UK 2016.
Published online: [14] F. X. Bon de Saint Hilaire, Philos. Trans. R. Soc. London 1710, 27, 2.
[15] A Midsummer Night’s Dream. The Arden Shakespeare, 2nd series, [45] J. Perez-Rigueiro, C. Viney, J. Llorca, M. Elices, J. Appl. Polym. Sci.
(Ed: H. F. Brooks), Methuen & Co. 1979, ISBN 0-415-02699-7. 2000, 75, 1270.
[16] M. Cartwright, Ancient History Encyclopedia, https://www.ancient. [46] H. W. Kwak, J. E. Ju, M. Shin, C. Holland, K. H. Lee, Biomacromol-
eu/Silk/ (accessed: February 2018). ecules 2017, 18, 2343.
[17] I. L. Good, J. M. Kenoyer, R. H. Meadow, Archaeometry 2009, [47] a) C. Z. Zhou, F. Confalonieri, M. Jacquet, R. Perasso, Z. G. Li,
51, 457. J. Janin, Proteins 2001, 44, 119; b) C. Z. Zhou, F. Confalonieri,
[18] A. Hyman, Science and Reform: Selected Works of Charles Babbage, N. Medina, Y. Zivanovic, C. Esnault, T. Yang, M. Jacquet, J. Janin,
Cambridge University Press, Cambridge, England 1989. M. Duguet, R. Perasso, Z. G. Li, Nucleic Acids Res. 2000, 28, 2413.
[19] FAOSTAT Statistics Devision, Food and Agriculture Organization [48] a) T. Asakura, K. Okushita, M. P. Williamson, Macromol-
of the United Nations, http://faostat3.fao.org/download/Q/QP/E ecules 2015, 48, 2345; b) S. W. Ha, H. S. Gracz, A. E. Tonelli,
(accessed: October 2016). S. M. Hudson, Biomacromolecules 2005, 6, 2563.
[20] T. M. Muffly, A. P. Tizzano, M. D. Walters, J. R. Soc. Med. 2011, [49] H. Yamada, Y. Igarashi, Y. Takasu, H. Saito, K. Tsubouchi, Biomate-
104, 107. rials 2004, 25, 467.
[21] a) A. Teshome, S. K. Raina, F. Vollrath, J. Insect Sci. 2014, 14, 36; [50] H. J. Jin, D. L. Kaplan, Nature 2003, 424, 1057.
b) M. F. Astudillo, G. Thalwitz, F. Vollrath, J. Cleaner Prod. 2014, [51] T. Wongpinyochit, B. Johnston, F. P. Seib, ACS Biomater. Sci. Eng.
81, 158. 2018, 4, 942.
[22] H. C. McCook, American Spiders and Their Spinningwork. A Natural [52] T. Asakura, K. Ohgo, T. Ishida, P. Taddei, P. Monti, R. Kishore, Bio-
History of the Orbweaving Spiders of the United States, with Special macromolecules 2005, 6, 468.
Regard to Their Industry and Habits, Vol. 3, Academy of Natural [53] E. Pennisi, R. F. Service, Science 2017, 358, 292.
Sciences of Philadelphia, Philadelphia, USA 1893. [54] L. Eisoldt, A. Smith, T. Scheibel, Mater. Today 2011, 14, 80.
[23] R. Lewis, BioScience 1996, 46, 636. [55] A. Sponner, W. Vater, S. Monajembashi, E. Unger, F. Grosse,
[24] S. Peers, Golden Spider Silk, V&A Publishing, London 2012. K. Weisshart, PLoS One 2007, 2, e998.
[25] A. Koeppel, C. Holland, ACS Biomater. Sci. Eng. 2017, 3, 226. [56] a) T. Lefevre, M. E. Rousseau, M. Pezolet, Biophys. J. 2007, 92,
[26] a) M. Tomenosuke, H. Saburo (Kanebo Ltd), Type US1714039A, 2885; b) J. D. van Beek, S. Hess, F. Vollrath, B. H. Meier, Proc.
1927; b) I. Wilhelm, M. Herbert (IG Farbenindustrie AG), Type Natl. Acad. Sci. USA 2002, 99, 10266; c) Q. Wan, K. J. Abrams,
US1913487A, 1930. R. C. Masters, A. C. S. Talari, I. U. Rehman, F. Claeyssens,
[27] C. R. Baumann, G. G. Diesser (Carl Rudolf BaumannGottlieb Got- C. Holland, C. Rodenburg, Adv. Mater. 2017, 29, 1703510.
tfried Diesser), Type US976977A, 1908. [57] a) S. Keten, Z. Xu, B. Ihle, M. J. Buehler, Nat. Mater. 2010, 9, 359;
[28] H. Masaru (SHOZABURO HOSHINO), Type US1603080A, 1924. b) K. Numata, Polym. J. 2015, 47, 537.
[29] F. Heinrich, R. Ernst, Type US1990588A, 1928. [58] a) C. Y. Hayashi, R. V. Lewis, J. Mol. Biol. 1998, 275, 773;
[30] L. D. Myers, L. A. Stegemeyer (TWITCHELL PROCESS Co), Type b) O. N. Tretinnikov, Y. Tamada, Langmuir 2001, 17, 7406.
US1896494A, 1929. [59] a) T. Asakura, M. Okonogi, K. Horiguchi, A. Aoki, H. Saito,
[31] A. P. Furman (Ira Furman), Type US1992202A, 1932. D. P. Knight, M. P. Williamson, Angew. Chem., Int. Ed. Engl.
[32] I. Meisel, R. Mülhaupt, Macromol. Chem. Phys. 2003, 204, 199. 2012, 51, 1212; b) T. Asakura, M. Okonogi, K. Horiguchi, A. Aoki,
[33] A. Bloch, A. S. Messores (Ethicon Inc), Type US3424164A, 1966. H. Saito, D. P. Knight, M. P. Williamson, Angew. Chem. 2012, 124,
[34] M. Iwatsuki, T. Hayashi (Ajinomoto Co Inc), Type US4818291A, 1238
1986 [60] a) R. E. Marsh, R. B. Corey, L. Pauling, Biochim. Biophys. Acta 1955,
[35] Y. Tsukada, N. Minoura (National Institute of Advanced Industrial 16, 1; b) Y. Takahashi, M. Gehoh, K. Yuzuriha, Int. J. Biol. Mac-
Science and Technology Minister of Agriculture, Forestry and Fish- romol. 1999, 24, 127.
eries), Type JPH01118544A, 1987. [61] a) D. L. Kaplan, C. M. Mello, S. Arcidiacono, S. Fossey, K. Senecal,
[36] T. Ninagawa, K. Tsubouchi (National Institute of Sericultual and W. Muller, in Protein-Based Materials, Springer, Boston 1997,
Entomological Science), Type JPH11104228A, 1997. p. 103; b) E. M. Pritchard, D. L. Kaplan, Expert Opin. Drug Delivery
[37] a) M. Murase (Marie Murase), Type JPH06166850A, 1992; 2011, 8, 797; c) K. Numata, P. Cebe, D. L. Kaplan, Biomaterials
b) S. Yoshida, K. Yamaura, S. Matsuzawa, M. Suzuki, Type 2010, 31, 2926.
JPH0669485B2, 1988. [62] a) J. Guan, F. Vollrath, D. Porter, Biomacromolecules 2011, 12,
[38] H. Akai, T. Nagashima, S. Terauchi (Takayuki Nagashima), 4030; b) D. Porter, F. Vollrath, Biochim. Biophys. Acta 2012, 1824,
Type JPH08268905A, 1995. 785.
[39] a) J. Huang, C. Wong, A. George, D. L. Kaplan, Biomaterials 2007, [63] C. Boutry, T. A. Blackledge, J. Exp. Biol. 2010, 213, 3505.
28, 2358; b) D. L. Kaplan, R. Nazarov, G. Vunjak-Novakovic, [64] B. B. Mandal, A. Grinberg, E. S. Gil, B. Panilaitis, D. L. Kaplan,
L. Meinel (Tufts University, Massachusetts Institute of Tech- Proc. Natl. Acad. Sci. USA 2012, 109, 7699.
nology), Type US7842780B2, 2003 [65] S. Yodmuang, S. L. McNamara, A. B. Nover, B. B. Mandal,
[40] a) P. C. DeMuth, Y. Min, D. J. Irvine, P. T. Hammond, Adv. M. Agarwal, T. A. Kelly, P. H. Chao, C. Hung, D. L. Kaplan,
Healthcare Mater. 2014, 3, 47; b) H.-Y. Wang, Y.-Q. Zhang, Soft G. Vunjak-Novakovic, Acta Biomater. 2015, 11, 27.
Matter 2013, 9, 138; c) Y. Chen, Y. Lin, K. M. Davis, Q. Wang, [66] D. F. Williams, Biomaterials 2008, 29, 2941.
J. Rnjak-Kovacina, C. Li, R. R. Isberg, C. A. Kumamoto, J. Mecsas, [67] a) M. Jewell, W. Daunch, B. Bengtson, E. Mortarino, Ann. N.Y.
D. L. Kaplan, Sci. Rep. 2015, 5, 13708; d) C. Holland, A. E. Terry, Acad. Sci. 2015, 1358, 44; b) A. van Turnhout, C. J. J. Franke,
D. Porter, F. Vollrath, Polymer 2007, 48, 3388. E. J. C. Vriens-Nieuwenhuis, W. B. van der Sluis, J. Plast. Reconstr.
[41] G. H. Altman, F. Diaz, C. Jakuba, T. Calabro, R. L. Horan, J. S. Chen, Aesthetic Surg. 2018, 71, 644.
H. Lu, J. Richmond, D. L. Kaplan, Biomaterials 2003, 24, 401. [68] S. Franz, S. Rammelt, D. Scharnweber, J. C. Simon, Biomaterials
[42] D. N. Rockwood, R. C. Preda, T. Yucel, X. Wang, M. L. Lovett, 2011, 32, 6692.
D. L. Kaplan, Nat. Protoc. 2011, 6, 1612. [69] A. E. Thurber, F. G. Omenetto, D. L. Kaplan, Biomaterials 2015, 71, 145.
[43] a) Y. Liu, Z. Z. Shao, F. Vollrath, Nat. Mater. 2005, 4, 901; [70] F. P. Seib, AIMS Bioeng. 2017, 4, 239.
b) F. Vollrath, Nature 2010, 466, 319. [71] a) F. P. Seib, M. Herklotz, K. A. Burke, M. F. Maitz, C. Werner,
[44] S. Barabino, Y. Chen, S. Chauhan, R. Dana, Prog. Retinal Eye Res. D. L. Kaplan, Biomaterials 2014, 35, 83; b) F. P. Seib, M. F. Maitz,
2012, 31, 271. X. Hu, C. Werner, D. L. Kaplan, Biomaterials 2012, 33, 1017.
[72] M. F. Maitz, C. Sperling, T. Wongpinyochit, M. Herklotz, [104] R. De Vita, E. M. Buccheri, M. Pozzi, G. Zoccali, J. Exp. Clin. Cancer
C. Werner, F. P. Seib, Nanomedicine 2017, 13, 2633. Res. 2014, 33, 78.
[73] N. A. Fine, M. Lehfeldt, J. E. Gross, S. Downey, G. M. Kind, [105] T. P. Crowley, T. Collin, J. Plast. Reconstr. Aesthetic Surg. 2015, 68, 1629.
G. Duda, D. Kulber, R. Horan, J. Ippolito, M. Jewell, Plast. Reconstr. [106] D. Almesberger, N. Zingaretti, C. Di Loreto, S. Massarut,
Surg. 2015, 135, 339. A. Pasqualucci, P. C. Parodi, J. Plast. Reconstr. Aesthetic Surg. 2015,
[74] J. E. Gross, R. L. Horan, M. Gaylord, R. E. Olsen, L. D. McGill, 68, 870.
J. M. Garcia-Lopez, K. Biber, K. Barnico, I. Toponarski, G. Altman, [107] N. Karp, M. Choi, D. A. Kulber, S. Downey, G. Duda, G. M. Kind,
Plast. Reconstr. Surg. 2014, 134, 700e. M. L. Jewell, D. K. Murphy, M. R. Lehfeldt, N. Fine, Plast. Reconstr.
[75] Z. Jiao, Y. Song, Y. Jin, C. Zhang, D. Peng, Z. Chen, P. Chang, Surg. Glob. Open 2017, 5, e1327.
S. C. Kundu, G. Wang, Z. Wang, L. Wang, Macromol. Biosci. 2017, [108] M. F. Freshwater, J. Plast. Reconstr. Aesthetic Surg. 2014, 67, 1457.
17, 1700229. [109] a) C. Fontanini, I. Berti, L. Monasta, G. Longo, G. Ital. Dermatol.
[76] B. Panilaitis, G. H. Altman, J. Chen, H. J. Jin, V. Karageorgiou, Venereol. 2013, 148, 293; b) D. Y. Koller, G. Halmerbauer, A. Bock,
D. L. Kaplan, Biomaterials 2003, 24, 3079. G. Engstler, Pediatr. Allergy Immunol. 2007, 18, 335; c) G. Ricci,
[77] H. K. Soong, K. R. Kenyon, Ophthalmology 1984, 91, 479. A. Patrizi, B. Bendandi, G. Menna, E. Varotti, M. Masi, Br. J.
[78] C. Pecquet, Eur. J. Dermatol. 2013, 23, 767. Dermatol. 2004, 150, 127; d) G. Stinco, F. Piccirillo, F. Valent,
[79] E. Fuchs, Dtsch. Med. Wochenschr. 1955, 80, 36. Dermatology 2008, 217, 191; e) K. S. Thomas, L. E. Bradshaw,
[80] a) J. C. Celedon, L. J. Palmer, X. Xu, B. Wang, Z. Fang, S. T. Weiss, T. H. Sach, J. M. Batchelor, S. Lawton, E. F. Harrison, R. H. Haines,
Pediatrics 2001, 107, E80; b) C. M. Wen, S. T. Ye, L. X. Zhou, Y. Yu, A. Ahmed, H. C. Williams, T. Dean, N. P. Burrows, I. Pollock,
Ann. Allergy 1990, 65, 375. J. Llewellyn, C. Crang, J. D. Grundy, J. Guiness, A. Gribbin,
[81] a) M. Dewair, X. Baur, K. Ziegler, J. Allergy Clin. Immunol. 1985, 76, E. J. Mitchell, F. Cowdell, S. J. Brown, A. A. Montgomery,
537; b) W. Zaoming, R. Codina, E. Fernandez-Caldas, R. F. Lockey, U. K. D. C. T. N. s. C. T. Team, PLoS Med. 2017, 14,
J. Invest. Allergol. Clin. Immunol. 1996, 6, 237. e1002280.
[82] P. S. Lai, D. C. Christiani, Curr. Opin. Pulm. Med. 2013, 19, 152. [110] C. Schaunig, D. Kopera, Int. J. Dermatol. 2017, 56, 589.
[83] R. I. Kunz, R. M. Brancalhao, L. F. Ribeiro, M. R. Natali, Biomed. [111] A. Inoue, I. Ishido, A. Shoji, H. Yamada, Contact Dermatitis 1997,
Res. Int. 2016, 2016, 8175701. 37, 185.
[84] H. Xie, W. Yang, J. Chen, J. Zhang, X. Lu, X. Zhao, K. Huang, [112] G. Senti, L. S. Steinmann, B. Fischer, R. Kurmann, T. Storni,
H. Li, P. Chang, Z. Wang, L. Wang, Adv. Healthcare Mater. 2015, P. Johansen, P. Schmid-Grendelmeier, B. Wuthrich, T. M. Kundig,
4, 2195. Dermatology 2006, 213, 228.
[85] B. Kundu, S. C. Kundu, Biomaterials 2012, 33, 7456. [113] W. Zhang, L. Chen, J. Chen, L. Wang, X. Gui, J. Ran, G. Xu,
[86] L. Meinel, S. Hofmann, V. Karageorgiou, C. Kirker-Head, J. McCool, H. Zhao, M. Zeng, J. Ji, L. Qian, J. Zhou, H. Ouyang, X. Zou, Adv.
G. Gronowicz, L. Zichner, R. Langer, G. Vunjak-Novakovic, Healthcare Mater. 2017, 6, 1700121.
D. L. Kaplan, Biomaterials 2005, 26, 147. [114] E. S. Gil, B. Panilaitis, E. Bellas, D. L. Kaplan, Adv. Healthcare
[87] Y. Wang, D. D. Rudym, A. Walsh, L. Abrahamsen, H. J. Kim, Mater. 2013, 2, 206.
H. S. Kim, C. Kirker-Head, D. L. Kaplan, Biomaterials 2008, 29, [115] a) J. H. Lee, D. K. Kim, H. S. Park, J. Y. Jeong, Y. K. Yeon, V. Kumar,
3415. S. H. Bae, J. M. Lee, B. M. Moon, C. H. Park, Laryngoscope 2016,
[88] J. Haupt, J. M. Garcia-Lopez, K. Chope, BMC Vet. Res. 2015, 11, 58. 126, 2798; b) J. H. Lee, J. S. Lee, D. K. Kim, C. H. Park, H. R. Lee,
[89] J. Brown, C. L. Lu, J. Coburn, D. L. Kaplan, Acta Biomater. 2015, 11, Clin. Exp. Otorhinolaryngol. 2015, 8, 117.
212. [116] J. Kim, C. H. Kim, C. H. Park, J. N. Seo, H. Kweon, S. W. Kang,
[90] K. Numata, D. L. Kaplan, Biochemistry 2010, 49, 3254. K. G. Lee, Wound Repair Regener. 2010, 18, 132.
[91] U. Slotta, S. Hess, K. Spiess, T. Stromer, L. Serpell, T. Scheibel, [117] C. L. Hu, J. Y. Cui, F. Z. Ren, C. Peng, Int. J. Food Eng. 2008, 4.
Macromol. Biosci. 2007, 7, 183. [118] C. Martinez-Mora, A. Mrowiec, E. M. Garcia-Vizcaino, A. Alcaraz,
[92] K. Lundmark, G. T. Westermark, A. Olsen, P. Westermark, Proc. J. L. Cenis, F. J. Nicolas, PLoS One 2012, 7, e42271.
Natl. Acad. Sci. USA 2005, 102, 6098. [119] D. W. Kim, H. S. Hwang, D. S. Kim, S. H. Sheen, D. H. Heo,
[93] S. Gomes, K. Numata, I. B. Leonor, J. F. Mano, R. L. Reis, G. Hwang, S. H. Kang, H. Kweon, Y. Y. Jo, S. W. Kang, K. G. Lee,
D. L. Kapan, Biomacromolecules 2011, 12, 1675. J. Park, W. S. Eum, Y. J. Cho, S. Y. Choi, J. Microb. Biotechnol. 2012,
[94] L. Fernandez-Garcia, N. Mari-Buye, J. A. Barios, R. Madurga, 22, 494.
M. Elices, J. Perez-Rigueiro, M. Ramos, G. V. Guinea, D. Gonzalez- [120] C. E. Kim, J. H. Lee, Y. K. Yeon, C. H. Park, J. Yang, Sci. Rep. 2017,
Nieto, Acta Biomater. 2016, 45, 262. 7, 44364.
[95] L. S. Wray, X. Hu, J. Gallego, I. Georgakoudi, F. G. Omenetto, [121] J. P. Whitcher, M. Srinivasan, M. P. Upadhyay, Bull. W. H. O. 2001,
D. Schmidt, D. L. Kaplan, J. Biomed. Mater. Res., Part B 2011, 99, 89. 79, 214.
[96] I. Greving, C. Dicko, A. Terry, P. Callowc, F. Vollrath, Soft Matter [122] K. Jaudzems, G. Askarieh, M. Landreh, K. Nordling,
2010, 6, 4389. M. Hedhammar, H. Jornvall, A. Rising, S. D. Knight, J. Johansson,
[97] a) N. Savage, Nature 2016, 533, S10; b) J. Wapner, Nature 2016, J. Mol. Biol. 2012, 422, 477.
533, S13. [123] a) D. H. Kim, J. Viventi, J. J. Amsden, J. Xiao, L. Vigeland,
[98] M. W. Clemens, S. Downey, F. Agullo, M. R. Lehfeldt, Y. S. Kim, J. A. Blanco, B. Panilaitis, E. S. Frechette, D. Contreras,
G. M. Kind, H. Palladino, D. Marshall, M. L. Jewell, A. B. Mathur, D. L. Kaplan, F. G. Omenetto, Y. Huang, K. C. Hwang,
B. P. Bengtson, Plast. Reconstr. Surg. Glob. Open 2014, 2, e246. M. R. Zakin, B. Litt, J. A. Rogers, Nat. Mater. 2010, 9, 511;
[99] J. E. Gross, Plast. Reconstr. Surg. Glob. Open 2013, 1, e86. b) H. Tao, S. W. Hwang, B. Marelli, B. An, J. E. Moreau, M. Yang,
[100] B. Bengtson, R. A. Baxter, M. W. Clemens, D. Bates, Plast. Reconstr. M. A. Brenckle, S. Kim, D. L. Kaplan, J. A. Rogers, F. G. Omenetto,
Surg. Glob. Open 2014, 2, e182. Proc. Natl. Acad. Sci. USA 2014, 111, 17385.
[101] N. Karp, M. Choi, J. Ippolito, M. Lehfeldt, M. Jewell, N. Fine, Eur. J. [124] S. Hazra, S. Nandi, D. Naskar, R. Guha, S. Chowdhury,
Surg. Oncol. 2014, 40, 621. N. Pradhan, S. C. Kundu, A. Konar, Sci. Rep. 2016, 6, 21840.
[102] A. Kornstein, Plast. Reconstr. Surg. Glob. Open 2014, 2, e244. [125] L. J. Bray, K. A. George, S. L. Ainscough, D. W. Hutmacher,
[103] A. N. Kornstein, Plast. Reconstr. Surg. Glob. Open 2014, 2, e190. T. V. Chirila, D. G. Harkin, Biomaterials 2011, 32, 5086.
[126] B. D. Lawrence, Z. Pan, A. H. Liu, D. L. Kaplan, M. I. Rosenblatt, (Eds: H. S. Azevedo, R. M. P. da Silva), Woodhead Publishing,
Acta Biomater. 2012, 8, 3732. Cambridge, MA, USA 2018, p. 27.
[127] L. Jia, C. E. Ghezzi, D. L. Kaplan, J. Biomed. Mater. Res., Part B [148] M. Floren, C. Migliaresi, A. Motta, J. Funct. Biomater. 2016, 7, E26.
2016, 104, 431. [149] G. G. Leisk, T. J. Lo, T. Yucel, Q. Lu, D. L. Kaplan, Adv. Mater. 2010,
[128] C. E. Ghezzi, L. Q. Wang, I. Behlau, J. Rnjak-Kovacina, S. R. Wang, 22, 711.
M. H. Goldstein, J. B. Liu, J. K. Marchant, M. I. Rosenblatt, [150] N. Kojic, M. J. Panzer, G. G. Leisk, W. K. Raja, M. Kojic,
D. L. Kaplan, J. Appl. Biomater. Funct. Mater. 2016, 14, E266. D. L. Kapan, Soft Matter 2012, 8, 6897.
[129] a) J. Wu, J. Rnjak-Kovacina, Y. Q. Du, M. L. Funderburgh, [151] F. P. Seib, E. M. Pritchard, D. L. Kaplan, Adv. Funct. Mater. 2013,
D. L. Kaplan, J. L. Funderburgh, Biomaterials 2014, 35, 3744; 23, 58.
b) S. Wang, C. E. Ghezzi, R. Gomes, R. E. Pollard, [152] F. P. Seib, M. Tsurkan, U. Freudenberg, D. L. Kapan, C. Werner,
J. L. Funderburgh, D. L. Kaplan, Biomaterials 2017, 112, 1. ACS Biomater. Sci. Eng. 2016, 2, 2287.
[130] J. F. Maya-Vetencourt, D. Ghezzi, M. R. Antognazza, E. Colombo, [153] M. L. Lovett, X. Wang, T. Yucel, L. York, M. Keirstead, L. Haggerty,
M. Mete, P. Feyen, A. Desii, A. Buschiazzo, M. Di Paolo, D. L. Kaplan, Eur. J. Pharm. Biopharm. 2015, 95, 271.
S. Di Marco, F. Ticconi, L. Emionite, D. Shmal, C. Marini, [154] D. C. Hamilton, H. H. Shih, R. A. Schubert, S. A. Michie,
I. Donelli, G. Freddi, R. Maccarone, S. Bisti, G. Sambuceti, P. N. Staats, D. L. Kaplan, M. J. Fontaine, J. Tissue Eng. Regener.
G. Pertile, G. Lanzani, F. Benfenati, Nat. Mater. 2017, Med. 2017, 11, 887.
16, 681. [155] H. Nakamura, J. Fang, H. Maeda, Expert Opin. Drug Delivery 2015,
[131] J. Lee, S. H. Park, I. H. Seo, K. J. Lee, W. Ryu, Eur. J. Pharm. Biop- 12, 53.
harm. 2015, 94, 11. [156] S. Wilhelm, A. J. Tavares, Q. Dai, S. Ohta, J. Audet, H. F. Dvorak,
[132] K. Tsioris, W. K. Raja, E. M. Pritchard, B. Panilaitis, D. L. Kaplan, W. C. W. Chan, Nat. Rev. Mater. 2016, 1, 16014.
F. G. Omenetto, Adv. Funct. Mater. 2012, 22, 330. [157] E. Wenk, H. P. Merkle, L. Meinel, J. Controlled Release 2011, 150,
[133] a) P. Johansen, T. Storni, L. Rettig, Z. Y. Qiu, A. Der-Sarkissian, 128.
K. A. Smith, V. Manolova, K. S. Lang, G. Senti, B. Mullhaupt, [158] Y.-Q. Zhang, W.-D. Shen, R.-L. Xiang, L.-J. Zhuge, W.-J. Gao,
T. Gerlach, R. F. Speck, A. Bot, T. M. Kundig, Proc. Natl. Acad. Sci. W.-B. Wang, J. Nanopart. Res. 2007, 9, 885.
USA 2008, 105, 5189; b) C. M. Jewell, S. C. B. Lopez, D. J. Irvine, [159] V. Gupta, A. Aseh, C. N. Rios, B. B. Aggarwal, A. B. Mathur, Int. J.
Proc. Natl. Acad. Sci. USA 2011, 108, 15745. Nanomed. 2009, 4, 115.
[134] A. B. Li, J. A. Kluge, N. A. Guziewicz, F. G. Omenetto, D. L. Kaplan, [160] J. Kundu, Y. I. Chung, Y. H. Kim, G. Tae, S. C. Kundu, Int. J. Pharm.
J. Controlled Release 2015, 219, 416. 2010, 388, 242.
[135] X. Jiang, J. Zhao, S. Wang, X. Sun, X. Zhang, J. Chen, D. L. Kaplan, [161] F. P. Seib, G. T. Jones, J. Rnjak-Kovacina, Y. Lin, D. L. Kaplan, Adv.
Z. Zhang, Biomaterials 2009, 30, 4522. Healthcare Mater. 2013, 2, 1606.
[136] a) J. Rnjak-Kovacina, L. S. Wray, J. M. Golinski, D. L. Kaplan, Adv. [162] J. D. Totten, T. Wongpinyochit, F. P. Seib, J. Drug Targeting 2017,
Funct. Mater. 2014, 24, 2188; b) M. Zamani, M. Khafaji, M. Naji, 25, 865.
M. Vossoughi, I. Alemzadeh, N. Haghighipour, Sci. Rep. 2017, 7, [163] Y. Q. Zhang, Y. J. Wang, H. Y. Wang, L. Zhu, Z. Z. Zhou, Soft
4455. Matter 2011, 7, 9728.
[137] a) M. J. Imola, D. D. Hamlar, W. Shao, K. Chowdhury, S. Tatum, [164] T. Wongpinyochit, P. Uhlmann, A. J. Urquhart, F. P. Seib, Biomac-
Arch. Facial Plast. Surg. 2001, 3, 79; b) S. J. Min, X. Gao, C. M. Han, romolecules 2015, 16, 3712.
Y. Chen, M. Y. Yang, L. J. Zhu, H. P. Zhang, L. Liu, J. M. Yao, J. [165] Y. Tian, X. Jiang, X. Chen, Z. Shao, W. Yang, Adv. Mater. 2014, 26,
Biomater. Sci., Polym. Ed. 2012, 23, 97. 7393.
[138] J. C. Middleton, A. J. Tipton, Biomaterials 2000, 21, 2335. [166] K. Numata, A. J. Mieszawska-Czajkowska, L. A. Kvenvold,
[139] G. S. Perrone, G. G. Leisk, T. J. Lo, J. E. Moreau, D. S. Haas, D. L. Kaplan, Macromol. Biosci. 2012, 12, 75.
B. J. Papenburg, E. B. Golden, B. P. Partlow, S. E. Fox, [167] a) T. B. Aigner, E. DeSimone, T. Scheibel, Adv. Mater. 2018, 30,
A. M. Ibrahim, S. J. Lin, D. L. Kaplan, Nat. Commun. 2014, 5, 3385. 1704636; b) Z. Zhou, S. Zhang, Y. Cao, B. Marelli, X. Xia, T. H. Tao,
[140] A. Schneider, X. Y. Wang, D. L. Kaplan, J. A. Garlick, C. Egles, Acta Adv. Mater. 2018, 30, 1706983.
Biomater. 2009, 5, 2570. [168] N. Dinjaski, D. L. Kaplan, Curr. Opin. Biotechnol. 2016, 39, 1.
[141] K. H. Kim, L. Jeong, H. N. Park, S. Y. Shin, W. H. Park, S. C. Lee, [169] K. Numata, D. L. Kaplan, Adv. Drug. Delivery Rev. 2010, 62, 1497.
T. I. Kim, Y. J. Park, Y. J. Seol, Y. M. Lee, Y. Ku, I. C. Rhyu, S. B. Han, [170] B. An, T. M. DesRochers, G. Qin, X. Xia, G. Thiagarajan,
C. P. Chung, J. Biotechnol. 2005, 120, 327. B. Brodsky, D. L. Kaplan, Biomaterials 2013, 34, 402.
[142] H. Fan, H. Liu, S. L. Toh, J. C. Goh, Biomaterials 2009, 30, 4967. [171] R. Price, A. Poursaid, H. Ghandehari, J. Controlled Release 2014,
[143] a) V. Catto, S. Fare, I. Cattaneo, M. Figliuzzi, A. Alessandrino, 190, 304.
G. Freddi, A. Remuzzi, M. C. Tanzi, Mater. Sci. Eng., C 2015, [172] X. Hu, X. Wang, J. Rnjak, A. S. Weiss, D. L. Kaplan, Biomaterials
54, 101; b) B. Marelli, A. Alessandrino, S. Fare, G. Freddi, 2010, 31, 8121.
D. Mantovani, M. C. Tanzi, Acta Biomater. 2010, 6, 4019; [173] J. O. Buitrago, K. D. Patel, A. El-Fiqi, J. H. Lee, B. Kundu,
c) E. C. Filipe, M. Santos, J. Hung, B. S. L. Lee, N. Yang, H. H. Lee, H. W. Kim, Acta Biomater. 2018, 69, 218.
A. H. P. Chan, M. K. C. Ng, J. Rnjak-Kovacina, S. G. Wise, J. Am. [174] M. M. Jacobsen, D. Li, N. Gyune Rim, D. Backman, M. L. Smith,
Coll. Cardiol. 2018, 3, 38. J. Y. Wong, Sci. Rep. 2017, 7, 45653.
[144] a) S. Kapoor, S. C. Kundu, Acta Biomater. 2016, 31, 17; [175] M. Farokhi, F. Mottaghitalab, S. Samani, M. A. Shokrgozar,
b) F. P. Seib, Ther. Delivery 2018, 9, 469. S. C. Kundu, R. L. Reis, Y. Fatahi, D. L. Kaplan, Biotechnol. Adv.
[145] R. D. Abbott, E. P. Kimmerling, D. M. Cairns, D. L. Kaplan, ACS 2018, 36, 68.
Appl. Mater. Interfaces 2016, 8, 21861. [176] J. Chang, X. F. Peng, K. Hijji, J. Cappello, H. Ghandehari,
[146] E. Bellas, T. J. Lo, E. P. Fournier, J. E. Brown, R. D. Abbott, E. S. Gil, S. D. Solares, J. Seog, J. Am. Chem. Soc. 2011, 133, 1745.
K. G. Marra, J. P. Rubin, G. G. Leisk, D. L. Kaplan, Adv. Healthcare [177] J. Jiang, S. Zhang, Z. Qian, N. Qin, W. Song, L. Sun, Z. Zhou,
Mater. 2015, 4, 452. Z. Shi, L. Chen, X. Li, Y. Mao, D. L. Kaplan, S. N. Gilbert Corder,
[147] F. P. Seib, in Self-Assembling Biomaterials: Molecular Design, X. Chen, M. Liu, F. G. Omenetto, X. Xia, T. H. Tao, Adv. Mater.
Characterization and Application in Biology and Medicine, 1st ed. 2018, 30, e1705919.
[178] M. Humenik, T. Scheibel, J. Phys.: Condens. Matter 2014, 26, T. Kanda, E. Abraham, M. Kamba, N. Komoto, J. L. Thomas,
503102. B. Mauchamp, G. Chavancy, P. Shirk, M. Fraser, J. C. Prudhomme,
[179] J. T. Prince, K. P. McGrath, C. M. DiGirolamo, D. L. Kaplan, Bio- P. Couble, Nat. Biotechnol. 2000, 18, 81.
chemistry 1995, 34, 10879. [197] J. R. dos Santos-Pinto, G. Lamprecht, W. Q. Chen, S. Heo,
[180] O. S. Rabotyagova, P. Cebe, D. L. Kaplan, Macromol. Biosci. 2010, J. G. Hardy, H. Priewalder, T. R. Scheibel, M. S. Palma, G. Lubec, J.
10, 49. Proteomics 2014, 105, 174.
[181] D. Huemmerich, T. Scheibel, F. Vollrath, S. Cohen, U. Gat, S. Ittah, [198] X. X. Xia, Z. G. Qian, C. S. Ki, Y. H. Park, D. L. Kaplan, S. Y. Lee,
Curr. Biol. 2004, 14, 2070. Proc. Natl. Acad. Sci. USA 2010, 107, 14059.
[182] Z. Megeed, M. Haider, D. Li, B. W. O’Malley Jr., J. Cappello, [199] A. M. Edlund, J. Jones, R. Lewis, J. C. Quinn, Nat. Biotechnol. 2018,
H. Ghandehari, J. Controlled Release 2004, 94, 433. 42, 12.
[183] S. Yanagisawa, Z. Zhu, I. Kobayashi, K. Uchino, Y. Tamada, [200] V. J. Venditto, F. C. Szoka Jr., Adv. Drug Delivery Rev. 2013, 65, 80.
T. Tamura, T. Asakura, Biomacromolecules 2007, 8, 3487. [201] a) R. F. Service, Science 2017, 358, 293; b) L. DeFrancesco, Nature
[184] a) C. Wong Po Foo, S. V. Patwardhan, D. J. Belton, B. Kitchel, Biotechnol. 2017, 35, 496.
D. Anastasiades, J. Huang, R. R. Naik, C. C. Perry, D. L. Kaplan, [202] M. Xu, R. V. Lewis, Proc. Natl. Acad. Sci. USA 1990, 87, 7120.
Proc. Natl. Acad. Sci. USA 2006, 103, 9428; b) L. L. Brott, R. R. Naik, [203] A. Lazaris, S. Arcidiacono, Y. Huang, J. F. Zhou, F. Duguay,
D. J. Pikas, S. M. Kirkpatrick, D. W. Tomlin, P. W. Whitlock, N. Chretien, E. A. Welsh, J. W. Soares, C. N. Karatzas, Science
S. J. Clarson, M. O. Stone, Nature 2001, 413, 291. 2002, 295, 472.
[185] K. Rittner, A. Benavente, A. Bompard-Sorlet, F. Heitz, G. Divita, [204] P. H. Zeplin, N. C. Maksimovikj, M. C. Jordan, J. Nickel, G. Lang,
R. Brasseur, E. Jacobs, Mol. Ther. 2002, 5, 104. A. H. Leimer, L. Römer, T. Scheibel, Adv. Funct. Mater. 2014, 24,
[186] S. Christian, J. Pilch, M. E. Akerman, K. Porkka, P. Laakkonen, 2658.
E. Ruoslahti, J. Cell Biol. 2003, 163, 871. [205] a) C. Muller, M. Hamedi, R. Karlsson, R. Jansson, R. Marcilla,
[187] K. D. Hermanson, D. Huemmerich, T. Scheibel, A. R. Bausch, Adv. M. Hedhammar, O. Inganas, Adv. Mater. 2011, 23, 898;
Mater. 2007, 19, 1810. b) L. Nileback, J. Hedin, M. Widhe, L. S. Floderus, A. Krona,
[188] a) U. K. Slotta, S. Rammensee, S. Gorb, T. Scheibel, Angew. Chem., H. Bysell, M. Hedhammar, Biomacromolecules 2017, 18, 846;
Int. Ed. Engl. 2008, 47, 4592; b) U. K. Slotta, S. Rammensee, c) N. D. Shalaly, M. Ria, U. Johansson, K. Avall, P. O. Berggren,
S. Gorb, T. Scheibel, Angew. Chem. 2008, 120, 4668. M. Hedhammar, Biomaterials 2016, 90, 50; d) M. Widhe,
[189] A. Lammel, M. Schwab, U. Slotta, G. Winter, T. Scheibel, Chem. N. D. Shalaly, M. Hedhammar, Biomaterials 2016, 74,
Sus. Chem 2008, 1, 413. 256.
[190] E. Doblhofer, T. Scheibel, J. Pharm. Sci. 2015, 104, 988. [206] G. Askarieh, M. Hedhammar, K. Nordling, A. Saenz, C. Casals,
[191] M. B. Elsner, H. M. Herold, S. Muller-Herrmann, H. Bargel, A. Rising, J. Johansson, S. D. Knight, Nature 2010, 465,
T. Scheibel, Biomater. Sci. 2015, 3, 543. 236.
[192] M. B. Schierling, E. Doblhofer, T. Scheibel, Biomater. Sci. 2016, 4, 1515. [207] M. Andersson, Q. Jia, A. Abella, X. Y. Lee, M. Landreh,
[193] K. Numata, M. R. Reagan, R. H. Goldstein, M. Rosenblatt, P. Purhonen, H. Hebert, M. Tenje, C. V. Robinson, Q. Meng,
D. L. Kaplan, Bioconjugate Chem. 2011, 22, 1605. G. R. Plaza, J. Johansson, A. Rising, Nat. Chem. Biol. 2017,
[194] M. Boulet-Audet, C. Holland, T. Gheysens, F. Vollrath, Biomacro- 13, 262.
molecules 2016, 17, 3198. [208] N. Kronqvist, M. Sarr, A. Lindqvist, K. Nordling, M. Otikovs,
[195] F. Teule, Y. G. Miao, B. H. Sohn, Y. S. Kim, J. J. Hull, M. J. Fraser Jr., L. Venturi, B. Pioselli, P. Purhonen, M. Landreh, H. Biverstal,
R. V. Lewis, D. L. Jarvis, Proc. Natl. Acad. Sci. USA 2012, 109, 923. Z. Toleikis, L. Sjoberg, C. V. Robinson, N. Pelizzi, H. Jornvall,
[196] a) Y. Kambe, K. Yamamoto, K. Kojima, Y. Tamada, N. Tomita, H. Hebert, K. Jaudzems, T. Curstedt, A. Rising, J. Johansson, Nat.
Biomaterials 2010, 31, 7503; b) T. Tamura, C. Thibert, C. Royer, Commun. 2017, 8, 15504.

Holland 2018

Uploaded by

Copyright:

Available Formats

Holland 2018

Uploaded by

Document Information

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Holland 2018

Uploaded by

Copyright:

Available Formats

PROGRESS REPORT

The Biomedical Use of Silk: Past, Present, Future

spend some time considering the motiva-

Dr. C. Holland 2. Introduction

Adv. Healthcare Mater. 2018, 1800465

© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

of water that acts as a plasticizer) and formed

Table 3. Reported human clinical trials using silk.

the treatment of ocular conditions, including dry eye[120] and

Due to their transparent nature, silk films

The composite microneedle consists of a silk fibroin tip loaded

Figure 6. Silk-based devices for fracture fixation. A) scanning electron

expressing cells and increases in collagen,

Silk fibroin hydrogels have emerged as

Motif Amino acid sequence Function Reference

Company Location Silk Products Clinical trial(s)

You might also like