Received: 11 October 2022 Revised: 28 December 2022 Accepted: 29 December 2022

DOI: 10.1002/app.53619

RESEARCH ARTICLE

Tuning of renewable sponge-like polyurethane

physical-chemical and morphological properties using
the pullulan as a reactive filler

Lana S. Maia 1 | Anne Shayene C. de Bomfim 2 | Daniel M. de Oliveira 2 |

Fernanda R. Pinhati 1 | Monique O. T. da Conceição 3 | Hernane S. Barud 4 |
Simone A. Medeiros 5 | Derval S. Rosa 6 | Daniella R. Mulinari 3
1
Department of Chemistry and Environmental, State University of Rio de Janeiro (UERJ), Rio de Janeiro, Brazil
2
Department of Materials and Technology, School of Engineering and Science, São Paulo State University (UNESP), São Paulo, Brazil
3
Department of Mechanical and Energy, State University of Rio de Janeiro (UERJ), Rio de Janeiro, Brazil
4
Department of Biotechnology, Laboratory of Polymers and Biomaterials, University of Araraquara (UNIARA), Araraquara, Brazil
5
Chemical Engineering Department, Engineering School of Lorena, University of São Paulo, São Paulo, Brazil
6
Center for Engineering, Modeling, and Applied Social Sciences (CECS), Federal University of ABC (UFABC), Santo André, Brazil

Correspondence
Daniella R. Mulinari, Department of Abstract
Mechanical and Energy, State University This study aimed to evaluate the effect of pullulan (Pull) on a renewable poly-
of Rio de Janeiro (UERJ), Rio de Janeiro,
urethane sponge-like scaffold synthesized from the Pull mixture with the castor
Brazil.
Email: [email protected]; oil-based polyol and the prepolymer using the situ foaming method. The Pull
[email protected] contents on the sponge-like are 5, 10, and 20 wt.%. The samples were evaluated
by optical microscopy (OM), scanning electron microscopy (SEM), density, Fou-
Funding information
Fundação Carlos Chagas de Amparo à rier transform infrared spectroscopy (FTIR), contact angle, Thermogravimetry,
Pesquisa do Estado do Rio de Janeiro, X-ray diffraction analysis (XRD), and compression strength test, respectively.
Grant/Award Numbers: E-
26/010.101232/2018, E-
FTIR results indicated that Pull was hydrogen-bonded to PU foam chains and
26/010/002530/2019, E-26/210.450/2021; increased the sponge-like scaffolds density, inducing a decrease in average cell
FAPESP, Grant/Award Numbers: FAPESP size compared to the pristine PU, confirming that they act as nucleating agents.
2020/13703-3, FAPESP 2019/04269-0
The Pull addition improved PU foams' hydrophobicity and mechanical proper-
ties and caused a thermal stability median between Pull and pristine PU. Thus,
all renewable sponge-like scaffolds were hydrophobics and presented appropri-
ate mechanical behavior, exhibiting better physicochemical properties, and
appearing as promising candidates for biomedical applications.

KEYWORDS
castor polyol-based polyurethane, pullulan, reactive filler, renewable scaffold, sponge-like

1 | INTRODUCTION various biological functions (e.g., energetic and structural) in

the organisms and microorganisms that produce them. Most
Natural polysaccharides (NPSs) are a polymeric material natural polysaccharides are found in plants in the form of
class widely used in industries due to their unique physical energy reserves; they can be synthesized by algae, bacteria,
and chemical characteristics.1 NPSs are synthesized to fulfill fungi, and yeasts or extracted from the exoskeleton of

J Appl Polym Sci. 2023;140:e53619. wileyonlinelibrary.com/journal/app © 2023 Wiley Periodicals LLC. 1 of 13

https://doi.org/10.1002/app.53619
2 of 13 MAIA ET AL.

arthropods.2 Some of these polysaccharides such as starch, flexibility, biodegradability, and physicochemical properties.
cellulose, carrageenan, or alginate are industrially extracted Besides, it has a broad spectrum of prospective biomedical
on large scale for applications as stabilizers, excipients, thick- and clinical applications due to its biocompatibility and
eners, and gelling agents.3 They often show biocompatibility, biostability.26,27 When proposing the study of antibacterial
bioactivity, and biodegradability, sharing similar structures biomaterials for skin dressings, Liang et al.21 reported that
to the natural extracellular matrix.4,5 Among the several the properties of PU can be achieved by the chemical struc-
resources for the synthesis of NPSs, the microbial exopoly- ture and molar mass of the polyol, the constitution of hard
saccharides (EPSs) have attracted much attention scientific and soft segments, and synthetic methods.21,28 Thus, the
community given their potential for food, cosmetic, pharma- choice of their building blocks (which are used in the syn-
ceutical, and biomedical applications.6–8 thesis of PU as macrodiols, diisocyanates, and chain
Pullulan (Pull), classed, as Generally Regarded as Safe extenders) can be implemented to obtain biomimetic con-
(GRAS) by the Food and Drug Administration (FDA) in structs, which can mimic the native tissue in terms of
the USA, is one of the more essential EPSs biopolymers.9 mechanical, morphological, and surface properties.29
It is produced by the fungus Aureobasidium pullulans and Aligned to this, the incorporation of fillers into the
contains α-1,6-glycosidic bonded maltotriose units.10,11 PU polymer matrix can generate products with improved
Their alpha linkages configuration (α-1,4 and α-1,6 link- antibacterial properties, cell proliferation, and migra-
ages) is responsible for the structural flexibility and water tion.21,27 Członka et al.30 proposed the development of
solubility, providing them with unique thin film and fiber- bio-based polyurethane composite foams with improved
forming characteristics, which are similar to certain mechanical, thermal, and antibacterial properties. They
petrol-derived plastics.9,12,13 Besides, it has wound healing, reported that adding clove filler (an Agro-industrial waste
antibacterial activities, and high thermal resistance (until with excellent antioxidant, antimicrobial, and antifungal
200
C).6 Another important issue discussed for Pull is activity properties) significantly enhances PU foams' anti-
their well-known non-mutagenic, non-carcinogenic, non- bacterial properties, manufacturing remarkable antimi-
toxic, biodegradable, and bioadhesive properties configur- crobial PU composite foams.
ing as a potential biomaterial.7,14,15 In the literature, there are works on the improvement
An exciting property of polymeric materials is their abil- of the antibacterial properties of PU through the insertion
ity to be efficiently designed according to the desired three- of cellulose nanofiber,31 Ag immobilized lignin,32 chitosan-
dimensional structure, such as hydrogels, macroporous scaf- modified graphene oxide,33 levofloxacin-loaded mesoporous
folds, microspheres, or micro-molded matrices, films, silica microspheres/nano-hydroxyapatite.34 However, there
sponges, and foams.4,7,16–18 In this context, the employment is a lack of work evaluating the feasibility of pullulan as a
of Pull for these devices' fabrication is continuously increas- PU filler for biomedical applications. Considering their out-
ing due to its outstanding antimicrobial property, which standing biocompatibility, biostability, and antimicrobial
can act as carriers for oral administration of therapeutic properties, the investigation of Pull into PU matrix for bio-
drugs, osteosynthesis materials (plates, pins, screws), and medical application is necessary.35,36
scaffolds for tissue engineering.19–21 Dalgic et al.22 proposed The purpose work was to synthesize and characterize
the development of a natural origin bilayer pullulan-PHBV sponges-like polyurethane from castor oil-based polyol
scaffold for wound healing applications. They reported that using pullulan as a reactive filler with different amounts
the nanofibrous Pull membrane presented a smaller pore (0, 5, 10, and 20 wt%). The amount of Pull and its impact
size providing an efficient barrier against bacterial transmis- on sponges-like polyurethane physical-chemical and
sion and enabling optimum oxygen and water vapor trans- morphological properties were evaluated.
mission. Other important works were published by
Raychaudhuri et al.23 and Muthusamy et al.24 which both
proposed a review approaching the several applications of 2 | MATERIAL A ND METHODS
pullulan in the biomedical area. They reported the develop-
ment of pullulan-based formulations with excellent biocom- 2.1 | Materials
patibility and antimicrobial properties for cancer treatment
and tumor targeting, stem cell therapy, vaccine, and gene Pullulan (Pull, Hayashibara Inc., Okayama, Japan) was used
deliveries. as received. The analysis of molar mass and dispersity of
Regarding possible polymeric matrices, polyurethane commercial pullulan via GPC provided Mn = 120 kDa,
(PU) represents a significant class of synthetic polymers that Mw = 244 kDa, and Đ = 2.03. For the synthesis of polyure-
consist of formed urethane linkages between isocyanate and thane sponge, the company Polyurethane (Minas Gerais,
polyol through polyaddition polymerization.21,25 PU is an Brazil) provided the following reagents: the renewable cas-
advanced functional material with excellent mechanical tor oil-based polyol (Biopol L40H, 531 mg KOH g1) and
MAIA ET AL. 3 of 13

F I G U R E 1 Schematic representation of the adopted methodology: (a) the process of preparing pristine PU and their sponge-like
scaffolds with different contents of Pull, and (b) polymerization reactive to obtain PU sponges. [Color figure can be viewed at
wileyonlinelibrary.com]

isocyanate (Biopol p-MDI, 4,4-diphenylmethane diisocya- inquire about the surface pores' morphological aspect,
nate, average functionality equal to 2.6, average molar mass shape, and distribution. The surface morphology of the
equal to 349.9 g mol1, solvent-free and 30%–32% of NCO). sponge-like scaffolds was determined by scanning electron
microscopy (SEM) (model TR-3000, HITACHI Ldt., Tokyo,
Japan). The equipment operated at 5 kV and a tungsten
2.2 | Sponge-like scaffolds filament recorded the images, employing a low-vacuum
technique and secondary electron detector. ImageJ soft-
The Pull/PU sponge-like scaffolds were manufactured ware was used to analyze PU cells' pore sizes, with
using the situ foaming method.37 First, the pristine PU 100 measures made for each sample. The pristine PU and
manual mixing of castor oil-based polyol and prepolymer their Pull/PU sponge-like scaffolds were cut and weighed
was made respecting the proportion of 1:1 for 50 s at to determine the apparent density into cubic shapes
room temperature (27
C) in molds. After 24 h, sponges (10  10  10 mm3). Densities were determined, by divid-
were removed from the molds to guarantee their com- ing the weight of the specimens by the calculated volume
plete expansion, and then the samples were cut for the by a digital caliper (MTX 316119). The values presented
characterization tests. The Pull/PU sponge-like was syn- correspond to the average density determined for five spec-
thesized from the pullulan (Pull) mixture with the castor imens of each material, as proposed by Zanini et al.38,39
oil-based polyol, and after that, the prepolymer was
added to the mixture. The sequential steps for the sponge
were analog to the pristine PU sponge. Samples were 2.3.2 | Fourier transformed infrared
named PU + X% Pull, where X represents the percentage spectroscopy
of pullulan (5, 10, and 20 wt.%) inserted in the PU. The
PU + X% Pull sponge-like scaffolds were cut into small The chemical structures of the pullulan (Pull), pristine PU,
cuboids (10  10  10 mm3). Figure 1 shows a schematic and their sponge-like scaffolds were evaluated by attenuated
representation of the adopted methodology. total reflectance (ATR) spectroscopy in a Perkin Elmer
Spectrophotometer (model Spectrum 100) by a transmit-
tance method, covering wavenumbers from 4000 to
2.3 | Characterization 650 cm1 with 12 scans and a spectral resolution of 4 cm1.

2.3.1 | Optical microscopy, scanning

electron microscopy, and density 2.3.3 | Contact angle

Stereomicroscopy was obtained using a stereoscope micro- The contact angle analysis (WCA) of pristine PU and their
scope (SZ STMPRO, BEL Engineering, Monza, Italy) to sponge-like scaffolds was performed using a Ramé-Hart
4 of 13 MAIA ET AL.

contact angle goniometer model Phoenix Mini
mf  mi

W g g1 ¼ ð1Þ
(Succasunna, USA). 20 measurements using 5 μl of water mi
drops were realized to investigate the surface interaction
(hydrophobicity/wetting) of pristine PU and their sponge- mf  mi
Efficiency ð%Þ ¼  100 ð2Þ
like scaffolds and, thus, obtain the average contact angle. mf

Where:
2.3.4 | Thermal properties - mf is the final sample mass.
- mi is the initial sample mass.
The thermogravimetric analysis (TGA) was performed
based on ASTM E2250 for Pull, pristine PU, and PU
sponge-like containing different Pull concentrations, to 3 | RESULTS A ND DISCUSSION
verify the mass loss as a function of temperature and the
temperature of thermal degradation. TGA was performed 3.1 | Optical microscopy, scanning
on TG/DTA 6200 from SII Nanotechnology – SEIKO. electron microscopy, and density
Samples with 5–10 mg were placed in a platinum pan,
and alumina was used as reference material. The follow- In 3D scaffolds, the porosity and pore size are impor-
ing parameters were used: temperature range from 30 to tant parameters that directly affects their functionality
700
C, a heating rate of 10
C min1, and a nitrogen during biomedical applications. Porosity, permeability,
atmosphere with a flow rate of 100 ml min1. and the mechanical properties of the scaffold are cru-
cial in terms of cell ingrowth, cell growth and migra-
tion, and scaffold colonization. Scaffold porosity is
2.3.5 | X-ray diffractometry determined by closed and open pores of varying sizes,
shapes, spatial distributions, and mutual interconnec-
The samples' structural properties were analyzed by X-ray tion.40 These porous and interconnected networks are
diffraction with XDR 600 equipment from Shimadzu with essential for cell nutrition, proliferation, mass trans-
Cu-Kα radiation (λ = 1.5406 Å) and 2θ in the range port/migration, and organization, which guarantees
between 10
to 80
. The objective was to identify the crys- potential vascularization and the formation of new tis-
talline and amorphous phases. sues.41,42 It is worth mentioning that this morphologic
structure also facilitates mechanical interlocking
between the scaffolds and concerned tissue, leading to
2.3.6 | Mechanical properties the full recovery of the injured organs.41,43 Figure 2
evidence the morphology of pristine PU, sponge-like
Pristine PU foam and their sponge-like scaffolds were scaffolds, and their respective pore size distribution
submitted to the compression tests by an EMIC testing and density.
machine (model DL2000) with pneumatic claws with a The micrograph analysis of the PU foam evidenced a
load cell of 5 kN, following the standard ASTM D 695-15, porous surface with interstitial spaces and closed cells.
with 5 mm min1 to 20% deformation. Five specimens of The addition of Pull in the PU matrix reduced the pore
50  50  25 mm3 and positioned between the two par- size compared to pristine PU and, consequently,
allel plates for compression were tested. increased the number of pores per area, as shown in the
histograms (Figure 2). In the pristine PU scaffold (PU),
the average pore size was obtained at 628 ± 212 μm. The
2.3.7 | Water uptake test average pore size of PU + 5% Pull, PU + 10% Pull, and
PU + 20% Pull reduced to 358 ± 94, 127 ± 26, and 136
The water absorption capacity of PU and sponge-like ± 40 μm, respectively. Subsequently, a 43%, 80%, and
scaffolds was determined using a dynamic system in trip- 78% decrease in pore size was observed for Pull rein-
licate. The sponges' samples were dimensioned in forced scaffolds compared to pristine PU. In general, scaf-
10  10  10 mm3. Then, 0.30 g of material was folds presented open and interconnected pores with
immersed in 50 ml of water and submitted to manual agi- different sizes and irregular shapes, presenting character-
tation for 5, 10, 20, and 40 min, 1, 2, and 4 h. After the istics of porous foams with a rough and continuous sur-
contact time, the sorbent was removed and drained to face.44 These observations may be related to interactions
eliminate excess water for 1 min. Lastly, the sample was between PU chains and Pull structure and this discussion
weighed in a precision balance. Equations (1) and (2): will better approach in the next section ‘Fourier
MAIA ET AL. 5 of 13

F I G U R E 2 Scanning electron microscopy (SEM) and optical microscopy (OM) images, as well as the pore size distribution histograms
and density of sponge-like scaffolds and pristine PU of (a) PU, (b) PU + 5% Pull, (c) PU + 10% Pull, and (d) PU + 20% Pull. [Color figure
can be viewed at wileyonlinelibrary.com]

transformed infrared spectroscopy’ (‘FTIR’). When eval- Pull in the PU matrix, the greater the apparent density
uating the development of Fe3O4 HA/PU superpara- of the scaffold. This fact can be explained due to the
magnetic composite porous scaffolds for bone repair additives incorporated during the cross-linking process
application, Yan et al.45 observed that the prepared scaf- directly affecting the foam density and pore size.49 The
folds exhibited a uniform porous structure, with a pore Pull acts as a crosslinking agent in PU foams and as
size distribution of 100–600 μm, which is beneficial to nucleation sites for cell formation, and since there is a
both the cells and bone tissue ingrowth. In the literature, higher number of cells that start to nucleate at the
recent works have reported similar pore size distribution same time, there is less gas available for cell growth,
in PU scaffolds, highlighting materials with pore sizes resulting in smaller pore sizes.38 Thus, the insertion of
greater than 100 μm.46–48 Therefore, the obtained PU Pull to the PU matrix increases the open cells, and this
+ X% Pull sponge-like scaffolds are configured as promis- behavior is more significant, mainly in higher contents
ing biomedical materials. (20 wt.%). Besides, the viscosity also increased during
Figure 2 evidenced that the pore size directly influ- the polymerization process due to the addition of filler,
ences the density values since the decreased pore size causing more centers of bubble nucleation than the
caused an increase in the density of the sponge-like pristine PU, influencing the morphology of the
scaffolds. The average density of the pristine scaffold sponges.
(PU) was obtained at 42 ± 1.1 g cm3. The average By single-factor ANOVA statistical analysis compar-
density of PU + 5% Pull, PU + 10% Pull, and PU ing the density (g cm3) versus the percentage of pullu-
+ 20% Pull enlarged to 45 ± 1.3, 47 ± 1.6, and 50 lan (0, 5, 10, and 20 wt.%) inserted in the PU, was
± 1.1 g cm3, respectively. The greater the amount of indicated a non-homogeneity of the variance due to a P-
6 of 13 MAIA ET AL.

value of less than 5%, 0.001 (Mean square = 42.59 and

Fvalue = 4.06), respectively, confirming the reliability of
the analysis, respectively, confirming the reliability of the
analysis.
The additives incorporated during the cross-linking
process directly affect the foam density and pores size. It
lowers the cell nucleation energy, promoting a higher
number of cells presenting a finer structure and smaller
size, resulting in a lower density foam.
Członka et al.30 observed a similar trend when study-
ing the clove-reinforced polyurethane composite foams
with antibacterial properties for biomedical applications.
It is worth mentioning that the relation between Pull's
dispersion and the PU matrix's viscosity is the main fac-
tor that influences the morphology of the sponge-like
scaffold. According to Martins et al.44 the viscosity
increases during the polymerization process due to the
adding filler, causing more centers of bubble nucleation
than the pristine PU. However, the increase in the nucle-
ation effect promoted an irregular morphology in the
sponge, composed of thinner cell walls.

3.2 | Fourier transformed infrared

spectroscopy

Fourier transformed infrared spectroscopy spectra of pris-

tine PU and Pull, PU + 5% Pull, PU + 10% Pull, and PU
+ 20% Pull are shown in Figure 3.
All samples showed typical infrared spectra of PU
foams: the presence of urethane bond could be observed F I G U R E 3 Fourier transformed infrared spectroscopy analysis
from peaks at 3490 cm1 (free N H), 3330 cm1 (hydro- (a) of pullulan, pristine PU, sponge like-scaffolds, and
gen-bonded N H), 1218 cm1 (C N) and 1064 cm1 deconvolution (b) of sponge-like scaffolds spectra. [Color figure can
(C O). The shoulder at 1615 cm1 evidenced the occur- be viewed at wileyonlinelibrary.com]
rence of urea linkages, and at 1710 cm1 is assigned to
the urethane carbonyl group. The characteristic absor-
bance of the N C O group appeared at 2277 cm1.50 was observed in pectin/pullulan films which hydroxyl
The FTIR spectrum of pullulan showed an intense group at 3272 cm1 reduced in intensity related to the
band at 3330 cm1, characteristic of hydroxyl groups. It pullulan peak, indicating the formation of intermolecular
was also identified characteristics peaks at 2923– H-bonds among the pullulan and pectin molecular
2898 cm1 (C H bonds), 1640 cm1 (O C O), chain.53 Besides, a shift was observed at the 1640 cm1
1354 cm1 (C O stretching), 1250–1070 cm1 (ether pullulan peak, considering the sponge-like scaffolds (this
bonds), 925 cm1 (a-1,6-glycosidic bonds), and 752 cm1 peak was not observed in the sponge-like scaffolds). In
(a-1,4-glycosidic bonds).51,52 general, in the FTIR of the sponge-like scaffolds PU,
Despite the overlapping of the OH and N-H bands in chemical groups with transformations in peak position or
all spectra, it is possible to notice an evident tendency to intensity were observed, in which is associated possible
reduce the band's intensity at 3330 cm1 in the sponge- bindings break and formation of new bonds between Pull
like scaffolds when compared to the intensity of the pull- and PU ‘s functional groups. To better understand the
ulan band. This behavior was more significative for scaf- peaks close to these displacements, the deconvolution at
folds reinforced with 15% and 20% Pull suggesting the 1710 cm1 peak of the sponge-like scaffolds was per-
possible physical interactions, such as H-bonds and van formed as represented in Figure 3 (b). This revealed the
der Walls interaction, between the urea group from PU presence of this peak at 1670 cm1 (PU + 5% Pull),
and the hydroxyl groups from pullulan.13 The same trend 1660 cm1 (PU + 10% Pull), and 1655 cm1 (PU + 20%
MAIA ET AL. 7 of 13

F I G U R E 4 Schematic representation of polyurethane (PU) foams and their sponge-like scaffolds considering possible hydrogen bonds
formed between PU chains and pullulan’ available active groups. [Color figure can be viewed at wileyonlinelibrary.com]

F I G U R E 5 Contact angle (WCA) of pristine PU and sponge-like scaffolds and a representative schematic of hydrophilic and
hydrophobic surfaces. [Color figure can be viewed at wileyonlinelibrary.com]

Pull). When evaluated the viability of preparation of understanding and control of the material characteristics
carboxymethyl-gellan and pullulan blend films, Zhang for regulating biocompatibility. One of the primary keys
et al.18 reported changes within the frequency range of to governing cell response is surface hydrophobicity,
1200 to 1700 cm1 and this trend is indicative of alter- which can be assessed by measuring the WCA through
ations in carboxymethyl structure. The authors noticed the water spread of a droplet on a surface.54 Figure 5
the shift of some peaks (2928, 1604, 1408 cm1) in the shows the measurements of the WCA for the pristine PU
spectra of carboxymethyl-gellan mixed with pullulan. and their respective sponge-like scaffolds.
This can suggest an interaction between the carbonyl The results indicated a hydrophobic nature both to
bonds from pristine PU and the pullulan (polysaccha- pristine PU and sponge-like scaffolds. Compared to the
rides) probably by a hydrogen bridge. pristine PU, the WCA values increased up to 1.9%, 10.6%,
To better understand the interactions between PU and 10.2% after adding Pull in order 5%, 10%, and 20%
and Pull, the proposed mechanism is elucidated in wt., respectively, possibly further increasing surface selec-
Figure 4. tivity. Due to an interaction between the carbonyl bonds
of pure PU and pullulan (polysaccharides) probably a
hydrogen bond was formed (as shown in Figure 4),
3.3 | Contact angle decreasing the pore size, conferring thus the water-
repellence property, that is, ‘lack of water affinity’. Water
Biomaterial development has focused on surface modifi- repellency is achieved decreasing the surface energy of
cations of polymeric matrixes to promote a greater the material, so that the water on the surface does not
8 of 13 MAIA ET AL.

T A B L E 1 Water absorption capacity and absorption efficiency TABLE 1 (Continued)

of pristine PU and sponge-like scaffolds as a function of time.
Water absorption after 4 h
Water absorption after 5 min
Absorption
Absorption Samples capacity (g g1) Efficiency (%)
Samples capacity (g g1) Efficiency (%) PU + 5% Pull 0.37 ± 0.04 27.0 ± 2.3
Pristine PU 0.054 ± 0.02 5.1 ± 1.5 PU + 10% Pull 0.43 ± 0.03 30.1 ± 1.3
PU + 5% Pull 0.12 ± 0.03 10.5 ± 2.3 PU + 20% Pull 0.48 ± 0.05 32.2 ± 2.3
PU + 10% Pull 0.079 ± 0.05 7.1 ± 4.7
PU + 20% Pull 0.24 ± 0.04 19.0 ± 3.0 “wet” the material. The adhesive force between the water
Water absorption after 10 min droplet and the sample surface decreases in the order
Absorption
80.98 < 78.71 < 70.09 < 68.23 mJ m2 for the pristine
Samples capacity (g g1) Efficiency (%) PU, PU + 5% Pull, PU + 10% Pull, and PU + 20% Pull
Pristine PU 0.12 ± 0.02 10.6 ± 1.5 sponges, respectively. It shows that with an increase in
PU + 5% Pull 0.29 ± 0.04 22.7 ± 2.4
hydrophobicity, the water molecules find it difficult to
get attached to the substrate surface.55 By single-factor
PU + 10% Pull 0.26 ± 0.01 20.7 ± 0.7
ANOVA statistical analysis comparing the CA values ver-
PU + 20% Pull 0.37 ± 0.02 27.0 ± 1.3
sus the percentage of pullulan (0, 5, 10, and 20 wt.%)
Water absorption after 20 min inserted in the PU, was indicated a non-homogeneity of
Absorption the variance due to a P-value of less than 5%, 0.0001
Samples capacity (g g1) Efficiency (%) (Mean square = 60.34 and Fvalue = 4.07), respectively,
Pristine PU 0.12 ± 0.02 10.6 ± 1.5 confirming the reliability of the analysis, respectively,
PU + 5% Pull 0.33 ± 0.05 25.1 ± 2.8 confirming the reliability of the analysis.
PU + 10% Pull 0.34 ± 0.003 25.7 ± 0.16 Table 1 shows water absorption capacity (g g1) and
PU + 20% Pull 0.43 ± 0.05 29.8 ± 2.4 efficiency (%) of neat PU and PU + X% Pull. Analyzing
the data, the scaffolds absorbed a slightly greater
Water absorption after 40 min
amount of water than the neat PU, and the insertion of
Absorption a high filler content in the PU resulted in a higher
Samples capacity (g g1) Efficiency (%)
water absorption value. Neat PU showed a maximum
Pristine PU 0.12 ± 0.02 10.6 ± 1.5
water uptake of 10.6% ± 1.5, while PU + 5% Pull, PU
PU + 5% Pull 0.40 ± 0.02 28.8 ± 1.3 + 10% Pull, and PU + 20% Pull scaffolds showed a
PU + 10% Pull 0.43 ± 0.03 30.1 ± 1.33 maximum water uptake of 28.8% ± 1.3, 30.1% ± 1.3,
PU + 20% Pull 0.48 ± 0.05 32.2 ± 2.3 and 32.2% ± 2.3, respectively, in 40 min. A similar
Water absorption after 1 h trend was observed by Atiqak et al.56 where the addi-
tion of palm fiber to PU increased water absorption
Absorption
Samples capacity (g g1) Efficiency (%) when compared to pure PU. This can be explained due
Pristine PU 0.12 ± 0.02 10.6 ± 1.5
to the pullulan consists of hydrophobic and hydro-
philic characteristics, which makes it a unique struc-
PU + 5% Pull 0.40 ± 0.02 28.8 ± 1.3
ture.17 This might favor water absorption since the
PU + 10% Pull 0.43 ± 0.03 30.1 ± 1.3
filler is on the surface of the polymeric matrix, how-
PU + 20% Pull 0.48 ± 0.05 32.2 ± 2.3
ever, with values low (lowest than 33%).
Water absorption after 2 h The hydrophobicity may be the result of the connec-
Absorption tions between the matrix and the reinforcement. When
Samples capacity (g g1) Efficiency (%) interacting with the available groups of hydrophilic char-
Pristine PU 0.12 ± 0.02 10.6 ± 1.5 acter (Pull), there was a decrease in their availability,
PU + 5% Pull 0.37 ± 0.04 27.0 ± 2.3 thus decreasing the hydrophilic tendency, resulting in a
PU + 10% Pull 0.43 ± 0.03 30.1 ± 1.3 hydrophobic material. This effect can be attributed to the
PU + 20% Pull 0.48 ± 0.05 32.2 ± 2.3 greater surface roughness of foams with smaller pore
sizes and the lack of large surface pores in which water
Water absorption after 4 h
droplets can be stored.49
Absorption Hydrophobic and superhydrophobic biomaterials are
Samples capacity (g g1) Efficiency (%)
materials with unique properties for biomedical applica-
Pristine PU 0.12 ± 0.02 10.6 ± 1.5 tions. In the review developed by Paterlini et al.57 was
MAIA ET AL. 9 of 13

F I G U R E 6 Weight (a) and derivative weight (b) TGA curves of the studied materials. [Color figure can be viewed at
wileyonlinelibrary.com]

cited that the success of a biomaterial depends on how decreases compared to pristine PU, increasing the stabil-
fluids, proteins, and cells interact with the foreign mate- ity of the scaffolds.
rial. Thus, the surface properties of biomaterial, such as Pristine PU showed a weight loss of 6.5% in the range
composition, charge, wettability, and roughness, must be of 100–250
C, possibly associated with a small portion of
optimized for the desired application to be achieved. unreacted reagents and a small amount of water retained
They affirmed also that hydrophobic surfaces can act as by the PU60 or related to the loss of volatile compounds
substrates to control protein adsorption, cellular interac- (additives used in the synthesis of PU).61 In the range of
tion, and drug delivery devices and diagnostic tools 250–440
C, a weight loss of 63.1% was also observed,
platforms. related to the scission of urethane bonds and loss of diphe-
nylmethane present in the prepolymer, followed by degra-
dation of the polyol (ester bonds) between 450–550
C with
3.4 | Thermal properties a weight loss of 11.7%.37,39,60,62,63
The scaffolds PU + 5% Pull presented a smaller
Thermogravimetric (TG) and first derivative thermogravi- weight loss percentage in the range of 225–440
C com-
metric (DTG) curves are shown in Figure 6, and the data pared to pristine PU, attributed to the incorporation of
are compiled in Table 2. filler (Pull) which decreases the active polyol groups,
Based on the results, Pull has at least one step of degra- reducing the reactive points. A minor weight loss was
dation and one primary thermal event (a slight first peak observed when inserting a more significant amount of
and a prominent peak in the DTG), whereas pristine PU pullulan into the PU.
and their sponge-like scaffolds show at least three steps The thermal properties of pullulan explain the impact of
(three peaks in the DTG). Pullulan presents a slight peak thermal degradation. The insertion of pullulan into PU
in the DTG curve between 30–150
C, with a 5% weight caused a higher residue percentage of 700
C (17.9%–26.7%)
loss and Tmax equals 77
C, related to moisture evaporation compared to pristine PU (15.5%) and pullulan (9.6%),
due to the inherent hydrophilicity.7,58 It also presents a evidencing intermediary thermal stability between pullulan
foremost degradation step between 280–450
C, with 74.2% and pristine PU. Anancharoenwong et al.64 observed similar
weight loss and Tmax equals 319
C, corresponding mainly behavior when evaluating natural rubber-based polyurethane
to the decomposition of the organic backbone of nanocomposites' thermo-mechanical and antimicrobial prop-
pullulan,7,59 and 9.6% residue at 700
C. erties for biomedical applications.
The TGA curves for pristine PU and sponge-like scaf-
folds showed similar behavior. On the other hand, when
inserting pullulan in polyurethane, the Tonset decreases 3.5 | X-ray diffractometry
compared to pristine PU (Tonset 250
C) but increases
compared to pullulan (Tonset 200
C), presenting an inter- Figure 7 evidence the physical characteristics of pullulan,
mediary behavior, which corroborates with data related pristine PU, and its sponge-like scaffolds obtained by the
to weight loss at 200
C. Besides, the amount of residue X-ray diffraction technique.
10 of 13 MAIA ET AL.

TABLE 2 Summary data from thermogravimetric analysis for pristine PU and their sponge-like scaffolds.

Weight loss (%)

Residue at Thermal Tmax Weight
Samples 100
C 200
C 300
C 400
C 500
C 700
C 700
C (%) Tonset (
C) event (
C) (
C) loss (
C)
Pull 3.4 5.4 13.6 80.3 83.7 90.4 9.6 200 30–150 77 5
280–450 319 74.2
Pristine PU 0 2.4 19.0 62.6 78.7 83.5 16.5 250 100–250 208 6.5
250–450 324 63.1
450–550 459 11.7
PU + 5% Pull 0 4.4 20.3 61.3 74.6 79.0 21.0 225 100–225 185 6
225–440 321 61.4
440–550 471 9.5
PU + 10% Pull 0 4.8 21.5 63.2 79.0 82.1 17.9 225 100–225 194 6.4
225–420 316 60.7
420–550 450 13.6
PU + 20% Pull 0 4.3 21.8 60.3 68.3 73.3 26.7 225 100–225 187 5.7
225–425 315 57.2
425–550 463 7.1

Abbreviations: Tmax, DTG peak temperature; Tonset, onset degradation temperature.

The X-ray diffractograms of pullulan showed a broad

peak appearing near 19
, corresponding to a molecular
packing distance of approximately 4.7 Å, indicating that
pullulan was a fully amorphous polymer. de Carvalho
et al.65 Islam et al.66 and Xiao et al.67 have reported similar
results. In addition, the X-ray diffractograms of pristine PU
and their sponge-like scaffolds showed a characteristic broad
peak at 20
, corresponding to a packing distance of approxi-
mately 4.4 Å. The addition of pullulan to PU showed that
the primary halo's position is visibly shifted compared to the
pullulan sample, indicating a decrease in the local molecu-
lar packing arrangement distance.68

3.6 | Mechanical properties

F I G U R E 7 X-ray diffraction (XRD) patterns of pullulan,
Mechanical properties are crucial for understanding the
pristine PU, and their sponge-like scaffolds. [Color figure can be
filler influence on scaffold structure, and these properties
viewed at wileyonlinelibrary.com]
can be strongly affected by the density of materials.62 The
effect of pullulan (Pull) on the polyurethane structure
was evaluated in terms of compressive strength and den- indicated a non-homogeneity of the variance due to a P-
sity, as shown in Figure 8. value of less than 5%, 0.00044 (Mean square = 0.0033 and
From Figure 8, it can be seen that pristine PU has a Fvalue = 4.07), respectively, confirming the reliability of the
compressive strength of 0.13 ± 0.01 MPa. A similar result analysis, respectively, confirming the reliability of the
was reported by Dzulkifli et al.69 Incorporating Pull into analysis.
the PU matrix, it can be observed that the biopolymer One hypothesis is that the polyurethane chains interact
exerted influence on the compressive strength values of with Pull improving the interfacial adhesion, corroborated
the sponge-like scaffolds with 5%, 10%, and 20% wt. Pull by the higher contact angles found in the sponge-like scaf-
increased compressive strength values of 30.8%, 34.6%, folds. A similar trend was reported by Czlonka et al.70 when
and 50.7%, respectively, compared to the pristine PU. studying the effect of clove in the PU matrix. They observed
By single-factor ANOVA statistical analysis comparing significant improvement in their materials' compressive
the compression strength values versus the percentage of strength after inserting 1% and 2% fiber. Another significant
pullulan (0, 5, 10, and 20 wt.%) inserted in the PU, was discussion is related to the density of scaffolds. According to
MAIA ET AL. 11 of 13

the literature, the incorporation of Pull can affect the cross- changed from an open-cell to a partial closed-cell structure
linking density of PU foams since the active groups as with a smaller pore size than neat PU, and composites
hydroxyls can be involved in the polymerization reaction, showed a higher contact angle. These additional crosslink-
increasing the number of urethane groups and hard seg- ing points could result in a PU matrix with dense structures,
ments. The presence of hard segments was confirmed in improving the mechanical properties of sponge-like
the FTIR section. Maia et al.37 evidenced similar behavior scaffolds.71,72
when evaluate PU composite foams with different coffee Although the results showed a low compressive
husk residues (CR) contents for sorption capacity to miti- strength when compared to cortical bone strength
gate oil contamination. It was observed that CR promoted (130–180 MPa),73 Keller et al.74 presented the compres-
harder segments within the PU structure, influencing the sive strength results for cancellous bones with values
composite foam's density. In addition, the pore morphology in the range of (0.1–16 MPa), which is in agreement
with the values obtained for the sponge-like scaffolds
samples. Therefore, the potential application would be
in the regeneration of small bone defects/voids or soft
tissue regeneration in vivo, as already reported by the
other authors in relation to polyurethane-based soft
foam.75
Figure 9 summarizes the main results and conclu-
sions presented in this work. This study creates an ideal
scenario for the emergence of innovative opportunities
using pullulan as well as the use of castor oil for the
development of sponge-like scaffolds. Considering all the
results, the pullulan incorporation in the polyurethane
foam resulted in materials with a porous structure (tiny
pores) and mechanical and thermal properties, which
could be applied in several biomedical areas.76 Sponge-
like with pullulan may offer solutions based on current
F I G U R E 8 Compressive tests and density of pristine PU, and challenges—using renewable raw materials instead of
their sponge-like scaffolds. [Color figure can be viewed at crude oil source resources and developing biomaterials
wileyonlinelibrary.com] that can help the world population's health.

FIGURE 9 Illustration of the main results obtained in this work. [Color figure can be viewed at wileyonlinelibrary.com]
12 of 13 MAIA ET AL.

4 | C ON C L U S I ON [3] F. G. Torres, O. P. Troncoso, A. Pisani, F. Gatto, G. Bardi, Int.

J. Mol. Sci. 2019, 20, 5092.
The pullulan effect on renewable castor polyol-based [4] N. Ahmed Omar, J. Amédée, D. Letourneur, J. C. Fricain, M.
Fenelon, Front. Bioeng. Biotechnol. 2022, 10, 1.
polyurethane sponge-like showed that the addition of
[5] M. Filippi, G. Born, M. Chaaban, A. Scherberich, Front.
Pull induces a decrease in average cell size compared to Bioeng. Biotechnol. 2020, 8, 474. https://doi.org/10.3389/fbioe.
the pristine PU, which increased the sponges-like density. 2020.00474
XRD patterns and FTIR data showed good interactions [6] M. Hamidi, O. V. Okoro, P. B. Milan, M. R. Khalili, H. Samadian,
between Pull and PU foam chains caused by hydrogen L. Nie, A. Shavandi, Carbohydr. Polym. 2022, 284, 119152.
bonds, improving mechanical properties. Furthermore, [7] M. Omar-Aziz, M. S. Yarmand, F. Khodaiyan, M. Mousavi, M.
the Pull incorporation into the PU caused intermediary Gharaghani, J. F. Kennedy, S. S. Hosseini, Int. J. Biol. Macro-
mol. 2020, 164, 3485.
thermal stability between Pull and pristine PU. The Pull
[8] R. S. Singh, N. Kaur, J. F. Kennedy, Carbohydr. Polym. 2015,
inserted into the castor-oil-derived polyurethane sponge-
123, 190.
like was hydrophobic and had proper mechanical behav- [9] M. Rai, M. Wypij, A. P. Ingle, J. Trzci nska-Wencel, P.
ior exhibiting better physicochemical characteristics com- Golinska, Int. J. Mol. Sci. 2021, 22, 13596. https://doi.org/10.
pared to pristine PU, making it a potential candidate for 3390/ijms222413596
biomedical applications. [10] R. S. Singh, N. Kaur, V. Rana, J. F. Kennedy, Carbohydr.
Polym. 2017, 171, 102.
A U T H O R C ON T R I B U T I O NS [11] S. Tang, L. Zhang, X. Mao, Y. Shao, M. Cao, L. Zhang, X.
Liang, Polym. Bull. 2022, 79, 10765.
Lana S. Maia: Methodology (lead); writing – original
[12] M. B. Coltelli, S. Danti, K. de Clerk, A. Lazzeri, P. Morganti,
draft (lead). Anne Shayene C. de Bomfim: Methodol- J. Funct. Biomater. 2020, 11, 1.
ogy (lead); validation (lead). Daniel M. de Oliveira: [13] S. Roy, P. Ezati, J. W. Rhim, ACS Appl. Bio Mater. 2022, 5,
Investigation (lead); methodology (lead); writing – review 2316.
and editing (lead). Fernanda R. Pinhati: Resources [14] A. Amrita, P. Sharma, D. S. Katti, Carbohydr. Polym. 2015,
(supporting); visualization (lead). Monique O. T. da 123, 180.
Conceição: Formal analysis (lead); investigation (lead); [15] T. Khaliq, M. Sohail, S. A. Shah, A. Mahmood, M. Kousar, N.
project administration (lead). Hernane S. Barud: Con- Jabeen, Int. J. Biol. Macromol. 2022, 209, 1826.
[16] H. Mert, B. Özkahraman, H. Damar, J. Drug Deliv. Sci. Tech-
ceptualization (supporting). Simone A. Medeiros: Con-
nol. 2020, 60, 101962.
ceptualization (lead); funding acquisition (supporting).
[17] R. S. Singh, N. Kaur, V. Rana, J. F. Kennedy, Carbohydr.
Derval S. Rosa: Funding acquisition (supporting); super- Polym. 2016, 153, 455.
vision (supporting). Daniella Mulinari: Funding acqui- [18] L. Zhang, J. Liu, X. Zheng, A. Zhang, X. Zhang, K. Tang, Car-
sition (lead); project administration (lead); resources bohydr. Polym. 2019, 216, 45.
(lead); supervision (lead); validation (lead); visualization [19] L. M. Ferreira, A. A. De Velasquez, S. R. Schaffazick, L. Cruz,
(lead); writing – review and editing (lead). Brazilian J. Pharm. Sci. 2015, 51, 27.
[20] T. Laube, J. Weisser, S. Berger, S. Börner, S. Bischoff, H.
Schubert, M. Gajda, R. Bräuer, M. Schnabelrauch, Mater. Sci.
A C K N O WL E D G M E N T S
Eng. C 2017, 78, 163.
This research was funded by Fundação Carlos Chagas de [21] Y. Liang, Y. Liang, H. Zhang, B. Guo, Asian J. Pharm. Sci.
Amparo à Pesquisa do Estado do Rio de Janeiro - 2022, 17, 353.
FAPERJ (E-26/010.101232/2018, E-26/010/002530/2019, [22] A. D. Dalgic, E. Koman, A. Karatas, A. Tezcaner, D. Keskin,
and E-26/210.450/2021), FAPESP 2020/13703-3 and Mater. Sci. Eng. C 2021, 134, 112554.
FAPESP 2019/04269-0. [23] R. Raychaudhuri, S. Naik, A. B. Shreya, N. Kandpal, A.
Pandey, G. Kalthur, S. Mutalik, Int. J. Biol. Macromol. 2020,
DATA AVAILABILITY STATEMENT 161, 1189.
[24] S. Muthusamy, S. J. Anandharaj, P. S. Kumar, Y. Meganathan,
The data that support the findings of this study are available
D. V. N. Vo, V. K. Vaidyanathan, S. Muthusamy, Microbial
from the corresponding author upon reasonable request.
Pullulan for Food, Biomedicine, Cosmetic, and Water Treat-
ment: A Review, Environmental Chemistry Letters, Vol. 20,
ORCID
Springer, New York 2022, pp. 3199-3234.
Daniella R. Mulinari https://orcid.org/0000-0003-4464- [25] L. Xu, X. Li, F. Jiang, X. Yu, J. Wang, F. Xiao, Constr. Build.
5620 Mater. 2022, 344, 128252.
[26] M. Gonzalez-Torres, I. H. Serrano-Aguilar, A. Cabrera-
R EF E RE N C E S Wrooman, R. Sanchez-Sanchez, R. Pichardo-Bahena, Y.
[1] Z. Wu, H. Li, X. Zhao, F. Ye, G. Zhao, Carbohydr. Polym. 2022, Melgarejo-Ramírez, G. Leyva-Gomez, H. Cortés, M. de los
284, 119182. Angeles Moyaho-Bernal, E. Lima, C. Ibarra, C. Velasquillo,
[2] D. Elsa, Polysaccharides 2022, 3, 480. Carbohydr. Polym. 2021, 270, 117916.
MAIA ET AL. 13 of 13

[27] G. Kasi, S. Gnanasekar, K. Zhang, E. T. Kang, L. Q. Xu, [55] J. Liu, B. Yu, B. Ma, X. Song, X. Cao, Z. Li, W. Yang, F. Zhou,
J. Appl. Polym. Sci. 2022, 139, 52181. Colloids Surf. A Physicochem. Eng. Asp. 2011, 380, 175.
[28] E. Shahrousvand, M. Shahrousvand, Mater. Today Commun. [56] A. Atiqah, M. Jawaid, M. R. Ishak, S. M. Sapuan, Procedia
2021, 27, 102421. Eng. 2017, 184, 581.
[29] S. Sartori, V. Chiono, C. Tonda-Turo, C. Mattu, G. Ciardelli, [57] T. T. Paterlini, L. F. B. Nogueira, C. B. Tovani, M. A. E. Cruz,
J. Mater. Chem. B 2014, 2, 5128. R. Derradi, A. P. Ramos, Biophys. Rev. 2017, 9, 683.
[30] S. Członka, A. Strakowska, K. Strzelec, A. Kairyte, A. [58] L. S. Martins, L. I. C. C. O. Cortat, N. C. Zanini, R. F. S.
Kremensas, Materials (Basel) 2020, 13, 1. Barbosa, A. G. Souza, S. F. Medeiros, D. S. Rosa, D. R.
[31] T. Y. Yu, Y. K. Tseng, T. H. Lin, T. C. Wang, Y. H. Tseng, Mulinari, Mater. Today Commun. 2021, 28, 102690.
Y. H. Chang, M. C. Wu, W. F. Su, Carbohydr. Polym. 2022, [59] S. Coseri, A. Spatareanu, L. Sacarescu, V. Socoliuc, I. S.
291, 119549. Stratulat, V. Harabagiu, J. Appl. Polym. Sci. 2016, 133, 2.
[32] H. Xie, H. Zhang, X. Liu, S. Tian, Y. Liu, S. Fu, Int. J. Biol. [60] D. I. Pelufo, S. C. Neto, R. C. B. Gobbo, A. J. dos Santos, A. J.
Macromol. 2021, 189, 690. Terezo, A. B. de Siqueira, J. Polym. Res. 2020, 27, 143. https://
[33] S. A. A. Najafabadi, A. Mohammadi, A. Z. Kharazi, Mater. Sci. doi.org/10.1007/s10965-020-02123-3
Eng. C 2020, 115, 110899. [61] J. M. Cangemi, S. C. Neto, G. O. Chierice, A. M. Dos Santos,
[34] Q. Wang, C. Chen, W. Liu, X. He, N. Zhou, D. Zhang, H. Gu, Polimeros 2006, 16, 129.
J. Li, J. Jiang, W. Huang, Sci. Rep. 2017, 7, 1. [62] B. P. de Oliveira, L. C. S. Balieiro, L. S. Maia, N. C. Zanini,
[35] L. C. Rusu, L. C. Ardelean, A. A. Jitariu, C. A. Miu, C. G. E. J. O. Teixeira, M. O. T. da Conceição, S. F. Medeiros, D. R.
Streian, Polymers (Basel) 2020, 12, 1. Mulinari, J. Mater. Cycles Waste Manag. 2022, 24, 553.
[36] S. Wendels, L. Avérous, Bioact. Mater. 2021, 6, 1083. [63] L. S. Martins, N. C. Zanini, L. S. Maia, A. G. Souza, R. F. S.
[37] L. S. Maia, N. C. Zanini, A. G. de Souza, R. F. S. Barbosa, D. S. Barbosa, D. S. Rosa, D. R. Mulinari, Chemosphere 2021, 267,
Rosa, H. S. da Barud, D. R. Mulinari, Iran. Polym. J. English 129288.
Ed. 2022, 31, 383. [64] E. Anancharoenwong, W. Chueangchayaphan, N. Rakkapao,
[38] N. C. Zanini, R. F. S. Barbosa, A. G. de Souza, D. S. Rosa, D. R. S. Marthosa, B. Chaisrikhwun, Polym. Bull. 2021, 78, 833.
Mulinari, J. Compos. Mater. 2021, 55, 813. [65] L. T. de Carvalho, M. L. da S. Paula, R. M. de Moraes, G. M.
[39] N. C. Zanini, A. G. de Souza, R. F. S. Barbosa, D. S. Rosa, D. R. Alves, T. M. Lacerda, J. C. Dos Santos, A. M. Dos Santos, S. de
Mulinari, J. Cell. Plast. 2021, 1–20, 139. F. Medeiros, Polymers (Basel) 2020, 12, 1.
[40] M. Bartoš, T. Suchý, R. Foltan, J. Geophys. Res. Planets 2018, 17, 1. [66] M. S. Islam, J. H. Yeum, A. K. Das, J. Colloid Interface Sci.
[41] Q. L. Loh, C. Choong, Tissue Eng. - Part B Rev. 2013, 19, 485. 2012, 368, 273.
[42] G. Lutzweiler, A. N. Halili, N. E. Vrana, Pharmaceutics 2020, [67] Q. Xiao, K. Lu, Q. Tong, C. Liu, J. Food Process Eng. 2015,
12, 1. 38, 155.
[43] I. Bružauskaitė, D. Bironaitė, E. Bagdonas, E. Bernotienė, [68] S. Bates, G. Zografi, D. Engers, K. Morris, K. Crowley, A.
Cytotechnology 2016, 68, 355. Newman, Pharm. Res. 2006, 23, 2333.
[44] L. S. Martins, F. M. Monticelli, D. R. Mulinari, Results Mater. [69] M. H. Dzulkifli, M. Y. Yahya, F. S. Md Akhir, R. A. Majid,
2020, 8, 100143. J. Teknol. Sciences Eng. 2014, 68, 53.
[45] Y. Yan, Y. Zhang, Y. Zuo, Q. Zou, J. Li, Y. Li, Mater. Lett. [70] S. Czlonka, A. Str ˛akowska, K. Strzelec, A. Kairytė, A.
2018, 212, 303. Kremensas, Materials 2020, 13, 1.
[46] H. Moris, A. Ghaee, M. Karimi, M. Nouri-Felekori, A. [71] S. Członka, A. Strąkowska, A. Kairytė, Materials (Basel) 2020, 13, 1.
Mashak, Biomater. Adv. 2022, 135, 212733. [72] H. Olcay, E. D. Kocak, Appl. Acoust. 2021, 173, 107733.
[47] M. Savaris, C. S. C. Garcia, M. Roesch-Ely, J. A. P. Henriques, [73] D. Bourell, D. Espalin, K. Arcaute, D. Rodriguez, F. Medina,
V. dos Santos, R. N. Brandalise, Mater. Sci. Eng. C 2019, M. Posner, R. Wicker, Rapid Prototyp. J. 2010, 16, 164.
96, 539. [74] T. S. Keller, D. R. Carter, C. J. Hernandez, G. S. Beaupre, Bone
[48] D. Xia, Y. Wang, R. Wu, Q. Zheng, G. Zhang, S. Xu, P. Zhou, 2001, 29, 74.
Appl. Mater. Today 2022, 29, 101607. [75] I. Gerges, M. Tamplenizza, F. Martello, C. Recordati, A.
[49] L. S. Maia, N. C. Zanini, A. M. Claro, N. C. Amaral, H. S. Tocchio, C. Lenardi, Acta Biomater. 2018, 73, 141.
Barud, D. R. Mulinari, J. Appl. Polym. Sci. 2021, 51259, [76] H. Zhang, Y. Jin, C. Chi, G. Han, W. Jiang, Z. Wang, H. Cheng,
51259. C. Zhang, G. Wang, C. Sun, Y. Chen, Y. Xi, M. Liu, X. Gao, X.
[50] X. Huang, C. F. De Hoop, J. Xie, Q. Wu, D. Boldor, J. Qi, Lin, L. Lv, J. Zhou, Y. Ding, Biomaterials 2021, 273, 120824.
Mater. Des. 2018, 138, 11.
[51] Z. Feng, S. Chen, A. Ahmad, L. Chen, W. Bai, Carbohydr.
Polym. 2022, 295, 119836.
[52] S. Li, J. Yi, X. Yu, Z. Wang, L. Wang, Int. J. Biol. Macromol. How to cite this article: L. S. Maia,
2020, 148, 258. A. S. C. de Bomfim, D. M. de Oliveira,
[53] R. Priyadarshi, S. M. Kim, J. W. Rhim, Food Chem. 2021, 347, F. R. Pinhati, M. O. T. da Conceição, H. S. Barud,
129022. S. A. Medeiros, D. S. Rosa, D. R. Mulinari, J. Appl.
[54] H.-I. Chang, Y. Wang, Regenerative Medicine and Tissue Engi-
Polym. Sci. 2023, 140(11), e53619. https://doi.org/
neering - Cells and Biomaterials. (Ed: D. EberliI), Intech Open,
London, UK 2011, Ch. 27, pp. 569. https://doi.org/10.5772/
10.1002/app.53619
21983