Polymer Testing 86 (2020) 106490

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Polymer Testing
journal homepage: http://www.elsevier.com/locate/polytest

Engineering quaternized chitosan in the 3D bacterial cellulose structure for

antibacterial wound dressings
Haiyong Ao a, *, Wenwen Jiang a, Yanjiao Nie a, Chen Zhou a, Jiajia Zong b, Mingzhuo Liu c,
Xuqiang Liu d, Yizao Wan a, e, *
a
Institute of Advanced Materials, East China Jiaotong University, Nanchang, 330013, China
b
School of Materials Science and Engineering, East China Jiaotong University, Nanchang, 330013, China
c
Department of Burn, The First Affiliated Hospital of Nanchang University, Nanchang, 330013, China
d
Department of Orthopedics, The First Affiliated Hospital of Nanchang University, The Artificial Joint Engineering and Technology Research Center of Jiangxi Province,
Nanchang, 330013, China
e
School of Materials Science and Engineering, Tianjin University, Tianjin, 300072, China

A R T I C L E I N F O A B S T R A C T

Keywords: Balancing antibacterial properties with biocompatibility is of paramount importance for wound dressings loaded
Wound dressings with antibacterial agents. In this work, a water soluble antibacterial agent, quaternized chitosan (hydrox­
Quaternized chitosan ypropyltrimethyl ammonium chloride chitosan, HACC) with an appropriate degree of substitution was intro­
Bacterial cellulose
duced into the bacterial cellulose (BC) network by adding it into the BC culture medium. Results indicated that
Antibacterial properties
Biocompotibility
the addition of HACC could affect the yield of BC, porous structure, thermal stability, water absorption and
antibacterial properties. HBC-1 with a low content of HACC (13.65 � 0.30%) cannot inhibit the biofilm for­
mation of bacteria, while HBC-3 with a high content of HACC (62.05 � 0.90%) has a low yield of BC and
confused structure. HBC-2 with an optimum concentration of HACC (37.33 � 0.80%) possessed a typical porous
structure, acceptable thermal stability, good water absorption and favorable antibacterial properties against
Staphylococcus aureus (S. aureus, ATCC 25923) and methicillin-resistant S. aureus (ATCC 43300). Most impor­
tantly, none of the HACC/BC films exhibited cytotoxicity to NIH3T3 cells. We believe that obtained HACC/BC
films with favorable bactericidal properties and biocompatibility could be potential candidates for wound
dressings in clinical applications.

1. Introduction long time. As inorganic antimicrobial agents, the antibacterial proper­

As a candidate for wound dressings, bacterial cellulose (BC) pro­
duced by several strains has a nanoscale three-dimensional (3D)
network structure that can retain moisture in wounds, ensure the cir­
culation of water and oxygen, remove excess exudates and prevent
bacterial invasion [1]. However, due to the lack of intrinsic bactericidal
performance, BC is unable to eliminate the colonized bacteria. There­
fore, much effort has been made to load antibacterial agents into the BC
network.
Antibiotics with broad-spectrum antibacterial activity, such as
chloramphenicol [2], ceftriaxone [3] and amoxicillin [4], were intro­
duced into a BC structure by adsorptive or covalent immobilization.
Nevertheless, drug resistance may appear after using antibiotics for a
ties of silver (Ag) and gold (Au) are as good as those of antibiotics [5,6].
Ag nanoparticle (AgNP)-deposited BC has excellent antibacterial effects
on Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) [7,8]. Ag
compounds such as Ag nitrate and Ag sulfadiazine (AgSD) have also
been used in burn and wound treatment [9,10]. However, the cytotox­
icity of Ag and Au may limit their clinical application on dressings [11,
12]. Zangeneh et al. found that after conjugation with extracts of plants
such as Camellia sinensis leaf [13], Hibiscus sabdariffa flower [14],
Thymus vulgaris leaf [15], and Spinacia oleracea L. leaf [16], the cyto­
toxicity of AgNPs and Au nanoparticles was reduced. In contrast, chi­
tosan (CS), possessing biocompatibility, nontoxicity, and intrinsic
antibacterial properties, is considered a favorable antibacterial material.
BC-CS films developed by Kingkaew et al. have been shown to be

* Corresponding author.
** Corresponding author. Institute of Advanced Materials, East China Jiaotong University, Nanchang, 330013, China.
E-mail addresses: aohyong@126.com (H. Ao), yzwan@tju.edu.cn (Y. Wan).

https://doi.org/10.1016/j.polymertesting.2020.106490
Received 5 January 2020; Received in revised form 8 March 2020; Accepted 12 March 2020
Available online 16 March 2020
0142-9418/© 2020 Elsevier Ltd. All rights reserved.
H. Ao et al.
effective against S. aureus and Aspergillus niger [17]. Lin et al. [18] found
that wounds treated with BC–CS epithelialized and regenerated faster
than those treated with BC. However, the poor water solubility of chi­
tosan affects its antimicrobial properties. Thus, it cannot be used to
address infected wounds in the clinical setting. Hence, there is a need to
select a suitable antimicrobial agent that simultaneously possesses
outstanding antibacterial properties and biocompatibility.
Hydroxypropyltrimethyl ammonium chloride chitosan (HACC),
which is a quaternized CS derivative, has drawn extensive attention as a
novel antimicrobial agent [19–22]. Due to its better water solubility,
HACC has significantly better antibacterial properties than CS [19,23].
Furthermore, in our previous work, we found that HACC-based multi­
layer modified titanium coatings could inhibit the colonization and
biofilm formation of several drug-resistant bacteria stains, including
methicillin-resistant S. aureus (MSRA, ATCC 43300) and
methicillin-resistant S. epidermidis (MRSE 287) isolated from the clinical
setting [24]. More importantly, HACC with a moderate degree of sub­
stitution (DS) of quaternary ammonium (approximately 18–26%)
clearly exhibits good biocompatibility with osteogenic cells [24–26].
Therefore, it is anticipated that loading HACC into the BC network may
endow BC dressings with antibacterial properties while maintaining
favorable biocompatibility. Nevertheless, introducing HACC into the BC
structure for skin dressings has not been reported.
The primary objective of the present study was to develop antimi­
crobial HACC/BC films for potential wound healing applications via in
situ biosynthesis. Scanning electron microscopy (SEM), Fourier trans­
form infrared spectroscopy (FTIR), X-ray diffraction (XRD) and ther­
mogravimetric (TG) analysis were used to characterize the obtained
HACC/BC films. The water absorption of the HACC/BC films was also
investigated. Furthermore, S. aureus (ATCC 25923) and ATCC 43300
and NIH3T3 fibroblast cells were employed to investigate the bacteri­
cidal activities and biocompatibility, respectively, of the HACC/BC
composite films.
2. Materials and methods
2.1. Preparation of the HACC/BC films
HACC/BC films were produced by the bacterial strain Acetobacter
xylinum X-2 via in situ preparation. HACC with 23% DS of quaternary
ammonium was synthesized according to a previous report [27]. HACC
was added into the BC culture medium at three HACC concentrations of
1, 3 and 5 mg/mL, respectively. After being sterilized at 115 � C for 30
min, the resultant three culture media were inoculated with the bacterial
strain. An aliquot of 2.8 mL of culture media containing HACC and
bacteria was added in a 24-well plate and stationary culture for 3 days.
The harvested HACC/BC samples were purified by ultrasonic oscillation
at 0.5% (w/v) NaOH solution for 0.5 h and were washed with deionized
water several times until they were neutral. Then, all samples were
freeze-dried and stored for later use. The obtained HACC/BC films were
named HBC-1, HBC-2 and HBC-3 corresponding to the HACC concen­
trations of 1, 3, and 5 mg/mL, respectively. Pure BC (pBC) membranes,
which served as a control group, were produced using the same pro­
cedure without adding HACC. The pBC and HACC/BC films were
weighed and the weight ratio of HACC was calculated.
2.2. Characterization of the HACC/BC films
The surface morphologies of the BC-based films were examined by
SEM (SU8010, Hitachi, Japan). The chemical compositions of the sam­
ples (pBC, HACC, HBC-1, HBC-2 and HBC-3) were investigated with
FTIR (DSQ II-TherMax 700 TGA-6700, Thermo Fisher, USA). The crys­
talline structure of the films was measured using XRD (D8 ADVANCE,
Bruker, Germany). The crystallinity index (C⋅I.) values were calculated
by using the Segal formula [28]. The water contact angles of pBC and
HACC/BC were measured using contact angle meter (Drop Master 300,
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Polymer Testing 86 (2020) 106490
Face-kyowa, Japan).
2.3. Pore size and porosity
The pore size measurements were carried out by measuring at least
100 randomly selected pores as reported in our previous work [29]. The
porosities of pBC and HACC/BC films were measured by the liquid
displacement method as described in a previous report [30].
Freeze-dried samples were immersed in absolute ethyl alcohol and the
porosity (P, %) was determined by the following equation:
W1 W0
P¼
ρV0
W0 and W1 represent the weight of the samples before and after
immersion, respectively. ρ is a constant of the density of absolute ethyl
alcohol and V0 is the volume of the sample before immersion.
2.4. Thermogravimetric analysis
The TG analysis of the samples was performed using Pyris 1 TG
analyzer (Diamond TG/DTA, PerkinElmer, USA) in the temperature
range of 50–800 � C at a constant heating rate of 10 � C/min.
2.5. Water absorption
Dried samples (Wdry) were allowed to absorb distilled water at room
temperature. At certain time intervals, after gently removing and aspi­
rating surface water by filter paper, the samples were weighed (Wwet).
The water absorption rate (WAR, %) of the samples was calculated as
follows:
Wwet Wdry
WAR ¼ � 100%
Wdry
2.6. Antibacterial assay
ATCC 25923 and ATCC 43300 were employed to investigate the
antibacterial properties of the BC and HACC/BC films. First, the disk
diffusion method was used to evaluate the antibacterial properties by
measuring the width of the zone of inhibition (ZOI). Typically, 200 μL of
bacterial suspensions with 1 � 108 colony-forming units (CFUs) was
evenly plated onto tryptone soy agar (TSA). A piece of a specimen was
placed on the center of the TSA and then cultivated in a thermostatic
biochemical incubator for 24 h at 37 � C. The width of ZOI was measured
by the following formula: H¼ (D-d)/2, where H is the width of the
bacteriostatic circle, D is the width of the outer diameter of the anti­
bacterial belt, and d is the diameter of each film.
The spread plate method was used to further study the antibacterial
properties and bacteriostatic rates of the HACC/BC films. One milliliter
of each bacterial suspension with 1 � 106 CFUs per mL was incubated
with a sample (pBC, HBC-1, HBC-2 and HBC-3) for 6 and 24 h. At the
designated time point, the adherent bacteria on the samples were dis­
lodged using ultrasonication. Then, the bacterial suspensions were
serially diluted 10-fold, plated onto TSA and incubated for 24 h. The
number of bacterial colonies on the substrates was counted and the
antibacterial rate of samples was calculated relative to the number of
bacterial colonies on BC. The number of bacterial colonies on pBC group
was measured as a baseline, and the bacteriostatic rates of HACC/BC
films were calculated by the microbial reduction compared to that of
pBC.
Regarding the SEM assay, after the samples were fixed in 2.5%
glutaraldehyde solution, the bacterial/material specimens were dehy­
drated using a series of graded ethanol solutions. Then, the samples were
observed through SEM.
H. Ao et al.
Fig. 1. SEM images of pBC and HACC/BC films.
2.7. Biocompatibility of the HACC/BC composite films
NIH3T3 cells were used to analyze the biocompatibility of the films
before and after introducing HACC. Briefly, sterilized materials were
placed in 24-well dishes containing 1 mL of cell suspension with 1 � 104
NIH3T3 cells and were cultured at 37 � C with 5% CO2 for 1, 3 and 6
days. The number of cells at each time point was analyzed with a CCK-8
assay kit according to the previously described procedure [31]. The OD
value of pBC group was measured as a baseline, and the cell viability of
NIH3T3 on HACC/BC films was expressed as the ratio of the OD value
relative to the value of the pBC group.
The growth of the cells on samples was also observed by SEM and
fluorescence microscope. After culturing cells with the specimens for 3
days, the cells were fixed in 2% glutaraldehyde overnight. The speci­
mens were rinsed twice in phosphate-buffered saline (PBS), followed by
dehydration in a graded series of ethanol and various ratios of ethanol/
hexamethyl disilazane (HMDS). Cells on the specimens were observed
by SEM. Regarding the fluorescence microscopy assay, cells were
stained with live/dead staining reagent (fluorescein diacetate and pro­
pidium iodide), and the morphologies of cells were visualized by
Fig. 2. The FTIR spectra (A) and XRD spectra (B) for pBC, HACC and HACC/BC films.
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Polymer Testing 86 (2020) 106490
fluorescence microscopy (TS2, Nikon, Japan).
2.8. Statistical analysis
All experiments were repeated three times. Statistical analysis was
performed with SPSS software. Variance analysis was used to compare
data between groups (ANOVA). p < 0.05 indicated a statistically sig­
nificant effect.
3. Results and discussion
3.1. Growth kinetics of BC
The components of the HACC/BC films are listed in Table S1. The
contents of HACC in the HACC/BC films were 2.80 mg for HBC-1, 8.40
mg for HBC-2 and 14.00 mg for HBC-3. After weighing the dry samples,
the contents of BC in the HACC/BC films were 17.76 � 1.05 mg for HBC-
1, 14.14 � 1.08 mg for HBC-2 and 8.58 � 0.71 mg for HBC-3. Compared
with the yield of pBC (18.44 � 0.99 mg), that of BC in the HACC/BC
films decreased. The decrease in the BC yield was only 3.68% for HBC-1
H. Ao et al.
Fig. 3. Typical thermal degradation profiles (A) and derivative curves (B) for pBC, HACC and HACC/BC films.
with 1 mg/mL HACC, and it rose to 23.32% for HBC-2 with 3 mg/mL
HACC. The largest decline occurred in HBC-3 (53.47%) with 5 mg/mL
HACC. The results indicated that as a mild antibacterial agent, HACC at
a low concentration has little effect on the biosynthesis of BC, but HACC
at a high concentration could seriously affect the output of BC. The
weight ratios of HACC were 13.65 � 0.30% for HBC-1, 37.33 � 0.80%
for HBC-2 and 62.05 � 0.90% for HBC-3.
3.2. Characterization of the HACC/BC composite films
The surface morphologies of the BC-based films were observed by
SEM. As shown in Fig. 1, the pBC film showed a typical 3D porous
nanofibrous structure, which is in agreement with our previous report
[32]. In contrast, BC fibers became less clear in HACC/BC films due to
the covering of BC fibers by HACC. It was very difficult to identify
regular BC fibers in the HBC-3 sample, as the HACC content was very
high (62.05 � 0.90%). Furthermore, the porosity and pore size of
HACC/BC films decreased with the increase in the HACC content, as
shown in Fig. S1.
To further characterize the obtained HACC/BC films, FTIR analysis
was employed. The FTIR spectra of the pBC, HACC and HACC/BC films
are shown in Fig. 2A. In the pBC spectrum, the peak at 3345 cm 1 was
due to the O–H stretching vibration, and the peaks at 2940 cm 1 and
2893 cm 1 were assigned to the aliphatic C–H stretching vibration. The
absorption peak at 1641 cm 1 was derived from the H–O–H bending of
water [33,34]. Moreover, an intense peak at 1148 cm 1 was due to the
C–O–C stretching vibration. In the HACC spectrum, two characteristic
peaks at 1651 cm 1 and 1570 cm 1 corresponded to the amide I ab­
sorption of the acetyl-glucosamine units (C– – O stretching) and the
deformation vibration of the amine groups (-NH2) of HACC, respectively
[35]. All characteristic bands of BC and HACC appeared in the spectra of
HACC/BC films. After careful examination, it can be found that
Fig. 4. Contact angle (A) and water absorption (B) of pBC and HACC/BC films.
4
Polymer Testing 86 (2020) 106490
characteristic peaks of C–H stretching, C– – O stretching and –NH2
deformation vibration were blueshifted. The results indicated that there
were intermolecular interactions between the BC fibrils and HACC
molecules.
The crystal structures of the pBC and HACC/BC films were deter­
mined through XRD measurement (Fig. 2B). The XRD pattern of pBC
exhibited diffraction peaks at 2θ values of approximately 14.76� and
22.37� , which were attributed to the (1 1 0) and (2 0 0) reflection planes
of the cellulose І structure, respectively [11,36,37]. After introducing
HACC, the XRD patterns of HBC-1, HBC-2 and HBC-3 did not change
significantly. The results showed that the introduction of HACC did not
influence the crystal structure of BC. Nevertheless, the C.I. values of
these samples are different. pBC shows the highest C.I. of 82.6%, while
the C.I. values were 80.6% for HBC-1, 79.1% for HBC-2 and 77.9% for
HBC-3, respectively. The decreasing trend of the C.I. with the content of
HACC is due to the intermolecular interaction between BC fibrils and
HACC molecules as elucidated by other researchers and our previous
studies [17,38–40].
3.3. Thermogravimetric analysis
TG analysis was used to investigate the thermal decomposition
behavior and thermal stabilities of HACC/BC films (Fig. 3). The pBC
films began decomposing at approximately 300 � C (Fig. 3A) and reached
a maximum decomposition rate at 360.9 � C, as shown in the derivative
TG curve (DTG) (Fig. 3B). The TG curves of HACC showed two mass loss
steps. The first step occurred from room temperature to 100 � C and
corresponded to a weight loss of 11%, which was mainly attributed to
the evaporation of free water adsorbed in HACC. The second mass loss
step, corresponding to the organic decomposition regions of HACC,
began at approximately 214 � C with the maximum peak at 252.8 � C in
the DTG curve. The final mass loss of 69% was at 800 � C, which was
H. Ao et al.
Fig. 5. Antibacterial properties of HACC/BC films. A: The viable bacteria on sample surfaces determined by the spread plate method after 6 h and 24 h incubation; B:
Bacteriostatic rates against S. aureus (ATCC 25923); C: Bacteriostatic rates against MRSA (ATCC 43300).
consistent with the literature [41]. The TG curves of the HACC/BC films
also showed two mass loss steps, the evaporation of free water and
organic decomposition. In the evaporation of free water step, the weight
loss was 8%. The organic decomposition began at 224–234 � C, which
was between the decomposition temperatures for HACC and pBC. Spe­
cifically, the maximum decomposition peaks were at 341.7 � C for
HBC-1, 331.4 � C for HBC-2, and 325.7 � C for HBC-3, which means that
the thermal stability of HACC/BC films decreased with the increase in
HACC.
3.4. Wettability and water absorption
Excellent absorbability is necessary for wound dressings to absorb
the excess exudates of wound in a timely manner [42]. The wettabilities
and water absorptions of the HACC/BC films were analyzed (Fig. 4). Due
to the abundant –OH groups on the BC fibers, the pBC film showed
excellent hydrophilicity and the contact angle of the pBC film was 38.0
� 2.5 � C (Fig. 4A). After HACC was introduced, the contact angles
increased to 47.0 � 2.4 � C for HBC-1, 59.0 � 2.8 � C for HBC-2 and 67.0
� 3.5 � C for HBC-3. The contact angle of HACC/BC films increased as the
content of HACC increased, because HACC was less hydrophilic than BC
and the contact angle of HACC was 82.0 � 3.4 � C.
Fig. 4B shows that the absorption rate of pBC was 60.46 � 3.64 g/g,
while the absorption rates of the three HACC/BC films were 40.74 �
2.03 g/g for HBC-1, 26.85 � 3.81 g/g for HBC-2, and 20.74 � 2.18 g/g
for HBC-3. The lower wettability of HACC may be one of the reasons for
these results. It was also reported that the wettability of the introduced
component could affect the water absorption capability of BC-based
composites. Lin et al. [18] found that the introduced CS could
decrease the absorption rate, while Saska et al. [43] found that the water
uptake of BC-based nanocomposites was improved by combining
collagen with excellent hydrophilicity. Another reason may be that the
intertwining of BC fibers limited the volume expansion of BC during
water absorption, while the introduced HACC led to a decrease in
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Polymer Testing 86 (2020) 106490
porosity, as described in Fig. S1. Therefore, the space for water ab­
sorption in HACC/BC films was less than that in pBC. Regarding
hydrogels with high water absorption, the water absorption will also be
decreased after the molecular chains are cross linked or form inter­
penetrating networks [44]. The water absorption rates of the HACC/BC
films were as good as that of BC, which may be helpful for removing the
tissue fluid of wounds in a timely manner.
3.5. Antibacterial assessment
The disk diffusion method was used to verify the antibacterial ac­
tivity of pBC and HACC/BC films against S. aureus and MRSA. Repre­
sentative images of ZOI around the samples are shown in Fig. S3. The
pBC group did not have a ZOI, which was consistent with previous re­
ports showing that pBC films did not have antibacterial properties [1].
All of the ZOIs of HBC-2 and HBC-3 against the two strains were very
obvious, and the widths of the ZOIs for HBC-2 were 2.7 � 0.5 mm
against S. aureus and 2.5 � 0.4 mm against MRSA, while those for HBC-3
were 2.5 � 0.4 mm against S. aureus and 2.4 � 0.4 mm against MRSA.
The results indicated that the HBC-2 and HBC-3 had satisfactory inhib­
itory effects against S. aureus and MRSA. Notably, the widths of the ZOIs
for HBC-1 were not clear against the two strains due to the low amount
of HACC.
The results of the spread plate method assay are shown in Fig. 5. It
can be found that the number of viable bacteria for each strain on the
three kinds of HACC/BC films was significantly lower than that on pBC
at 6 h (Fig. 5A), which indicated that all HACC/BC groups could reduce
pathogenic bacterial colonization of both the bacterial stains. After 24 h
of cultivation, the number of viable bacteria on HBC-2 and HBC-3 was
also few, while lots of viable bacteria were found on plates of the HBC-1
group. The results mean that HBC-2 and HBC-3 could inhibit the biofilm
formation of pathogenic bacterial but HBC-1 cannot. From the results of
the bacteriostatic rates (Fig. 5B and C), it can be seen that the bacte­
riostatic rates of all HACC/BC samples against the two strains were
H. Ao et al.
Fig. 6. Viability of NIH3T3 cells on the surface of pBC and HACC/BC films at 1,
3 and 6 days was expressed as the ratio of the OD value relative to the value
of pBC.
greater than 90% after incubation for 6 h. At 24 h, the bacteriostatic
rates of the HBC-1 group were approximately 29.4% against ATCC
25923 and approximately 27.2% against ATCC 43300, while those of
HBC-2 and HBC-3 were still over 90% at 24 h. These results indicated
that HBC-1 with 13.65 � 0.30% HACC did not have satisfactory anti­
bacterial properties, and with the increase in HACC, the bacteriostatic
activity of the HACC/BC films was improved.
The characterization of bacterial adhesion and biofilm formation on
surface of the HACC/BC samples was confirmed with SEM (Fig. S3). It
was observed that many bacteria adhered to the surface of the pBC film
at 6 h, while it was difficult to find bacteria on the three HACC/BC films.
Fig. 7. Fluorescence micrographs of NIH-3T3 cells with living/Dead cells staining after proliferation on the four different specimens for 3 days.
6
Polymer Testing 86 (2020) 106490
After incubation for 24 h, a thick bacterial biofilm formed on the surface
of BC, and many bacteria were seen on HBC-1, while few bacteria were
observed on HBC-2 and HBC-3, which agreed well with the results of the
antibacterial experiments.
3.6. Cell viability assay
The characterization of cell viability is crucial for potential wound
dressing materials [42]. To evaluate the biocompatibility of HACC/BC,
the viability of NIH3T3 on specimens was evaluated by a CCK-8 assay
and the results are shown in Fig. 6. At all time points, the viability of
NIH3T3 on all HACC/BC groups was as good as that of on pBC, there
were no statistically significant differences between them. The results
indicated that the introduction of HACC with 23% DS did not have a
negative influence on the cytocompatibility of the BC-based composite
films, although the percentages of HACC were high at 37.33 � 0.80% in
HBC-2 and 62.05 � 0.90% in HBC-3. This conclusion was also verified
by the results of live/dead staining assay (Fig. 7). After the cells were
cultured for 3 days, there was no dead cell on all specimens. The mor­
phologies of NIH-3T3 cells cultured for 3 days on the four different
specimens were observed by SEM (Fig. S4). It can be found that there
were many cells on the pBC and HACC/BC films. These results suggested
that the HACC/BC films had good biocompatibility although the content
of HACC was as high as 62.05 � 0.90% in HBC-3, which was consistent
with many previous reports. Peng et al. [27] showed that HACC with
18% DS was noncytotoxic to L-929 cells and had good biocompatibility
with osteogenic cells. The results reported by Tan et al. [45] indicated
that loaded HACC with 26% DS did not influence the proliferation of
hMSCs on HACC/polymethylmethacrylate. Yang et al. [46] cross-linked
H. Ao et al.
HACC onto PLGA nanofiber membranes, and the PLGA-HACC mem­
branes exhibited favorable biocompatibility and a significant ability to
stimulate the adhesion, spread and proliferation of human dermal fi­
broblasts and HaCaT cells.
4. Conclusions
In conclusion, HACC/BC films were fabricated via in situ biosyn­
thesis for wound dressings. Adding HACC to the culture medium of BC
could affect the growth kinetics, yield and crystallinity of BC, depending
on the content of HACC, and it was found that the content of HACC could
also affect the physicochemical and antibacterial properties. When an
optimum concentration of HACC (3 mg/mL) was introduced into the
culture medium, the obtained HACC/BC films, namely, HBC-2 with
37.33 � 0.80% HACC, possessed an acceptable yield of BC, typical
porous structure, and high water absorption. Most importantly, it
showed favorable antibacterial properties against S. aureus and MRSA
while still maintaining acceptable biocompatibility. Therefore, a good
HACC-loaded BC wound dressing with good antibacterial properties and
biocompatibility could be obtained by optimizing the content of HACC
with an appropriate DS. We believe that the optimized HBC-2 wound
dressing can meet the requirements for use in a clinical setting.
Data availibility statement
The processed data required to reproduce these findings would be
available on request.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
CRediT authorship contribution statement
Haiyong Ao: Conceptualization, Funding acquisition, Resources,
Writing - original draft, Project administration. Wenwen Jiang: Inves­
tigation, Data curation. Yanjiao Nie: Investigation. Chen Zhou: Inves­
tigation. Jiajia Zong: Formal analysis. Mingzhuo Liu: Writing - review
& editing. Xuqiang Liu: Methodology. Yizao Wan: Funding acquisition,
Resources, Writing - review & editing.
Acknowledgements
This work is supported by the National Natural Science Foundation
of China (Grant No. 31760265, 81501856, 31870963 and 51973058),
grant awarded by Natural Science Foundation of Jiangxi Province
(20192ACB80008, 20171ACB21036 and 20181BAB205019).
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.polymertesting.2020.106490.
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