Journal of Colloid and Interface Science 556 (2019) 514–528

Contents lists available at ScienceDirect

Journal of Colloid and Interface Science

journal homepage: www.elsevier.com/locate/jcis

Mussel-inspired, antibacterial, conductive, antioxidant, injectable

composite hydrogel wound dressing to promote the regeneration of
infected skin
Yongping Liang a, Xin Zhao a, Tianli Hu a, Yong Han a, Baolin Guo a,b,⇑
a
Frontier Institute of Science and Technology, and State Key Laboratory for Mechanical Behavior of Materials, Xi’an Jiaotong University, Xi’an 710049, China
b
Key Laboratory of Shaanxi Province for Craniofacial Precision Medicine Research, College of Stomatology, Xi’an Jiaotong University, Xi’an 710049, China

g r a p h i c a l a b s t r a c t

a r t i c l e i n f o a b s t r a c t

Article history: Infection is a major obstacle to wound healing. To enhance the healing of infected wounds, dressings with
Received 9 June 2019 antibacterial activities and multifunctional properties to promote wound healing are highly desirable.
Revised 22 August 2019 Herein, gelatin-grafted-dopamine (GT-DA) and polydopamine-coated carbon nanotubes (CNT-PDA) were
Accepted 23 August 2019
used to engineer antibacterial, adhesive, antioxidant and conductive GT-DA/chitosan/CNT composite
Available online 24 August 2019
hydrogels through the oxidative coupling of catechol groups using a H2O2/HRP (horseradish peroxidase)
catalytic system. The addition of the antibiotic doxycycline endowed the hydrogels with antimicrobial
Keywords:
activity to treat infected full-thickness defect wounds. Additionally, CNT-PDA endowed these hydrogels
Antibacterial adhesive hydrogel
Conductive hydrogel dressing
with an excellent photothermal effect, leading to good in vitro and in vivo antibacterial activities against
Hemostat Gram-positive and Gram-negative bacteria. The catechol group and polydopamine imparted tissue adhe-
Antioxidant siveness, and the hemostatic and antioxidant abilities of these hydrogels were also investigated. The
Infected wound healing porosity, degradability, swelling, rheological, mechanical, and conductive behaviors of these hydrogels
were finely regulated by changing the concentration of CNT-PDA. Hemolysis and cytocompatibility tests
using L929 fibroblast cells confirmed the good biocompatibility of these hydrogels. The wound closure,
collagen deposition, histomorphological examination and immunofluorescence staining results demon-
strated the excellent effects of these hydrogels in an infected full-thickness mouse skin defect wound.
In summary, the adhesive antibacterial and conductive GT-DA/chitosan/CNT hydrogels showed great
potential as multifunctional bioactive dressings for the treatment of infected wounds.
Ó 2019 Elsevier Inc. All rights reserved.

⇑ Corresponding author at: Frontier Institute of Science and Technology, and State
Key Laboratory for Mechanical Behavior of Materials, Xi’an Jiaotong University,
Xi’an 710049, China.
E-mail address: baoling@mail.xjtu.edu.cn (B. Guo).
1. Introduction
It is well-known that infection is a major obstacle to wound
healing [1], and it has become a continuously growing cause of

https://doi.org/10.1016/j.jcis.2019.08.083
0021-9797/Ó 2019 Elsevier Inc. All rights reserved.
 Y. Liang et al. / Journal of Colloid and Interface Science 556 (2019) 514–528
death among patients with serious illnesses. In addition, the treat-
ment of infection places a substantial burden on the medical sys-
tem and even society overall. Currently, antibiotics are still the
main strategy used in the clinical treatment of infection. Therefore,
developing proper antibacterial drug delivery systems that can not
only effectively and accurately deliver antibiotics to the wound site
but also control the release behaviors for active wound healing are
highly desirable [2]. Thus, many types of modern wound dressings
including semipermeable films, semipermeable foams, hydrocol-
loids and hydrogels with sustained drug release properties have
been developed in an attempt to not only avoid the infection of
defect wounds but also to promote the wound repair process [3–
6]. Among these dressings, due to their hydrophilic nature, hydro-
gels can reduce the risk of wound infection by absorbing wound
exudate and maintaining a moist environment, characteristics
which present good potential for improving the repair of damaged
tissue.
As a protein derivative, gelatin (GT) is considered beneficial for
wound healing due to its non-immunogenicity, cell adhesion
behavior and blood coagulation characteristics. GT has been used
in combination with a large number of synthetic or natural macro-
molecules to fabricate wound dressings. For example, the addition
of chitosan (CS) can overcome the shortcomings (weak mechanical
strength and fast degradation rate) of gelatin-based hydrogels. In
addition, CS itself can also trigger hemostasis and accelerate tissue
regeneration due to the migration of inflammatory cells and the
activation of fibroblasts that produce various cytokines [7]. More-
over, previous works on polyelectrolyte GT-CS scaffolds/films
[8,9] have demonstrated their excellent potential for use in skin
tissue engineering applications. On the other hand, excessive reac-
tive oxygen species (ROS) during the wound repair process often
affect the repair by changing or degrading extracellular matrix
(ECM) proteins, damaging dermal fibroblasts and reducing the
function of keratinocytes [10]. Therefore, controlling the levels of
ROS has been demonstrated to be an effective way to promote
wound healing [11]. However, hydrogels based on GT/CS have no
antioxidant capacity. Fortunately, the structure of catechol has
been proven to be widely present in a variety of natural antioxi-
dants and plays an important role in scavenging ROS [12]. Thus,
grafting dopamine to GT (gelatin-grafted-dopamine (GT-DA)) will
endow GT with good antioxidant properties [13,14]. Moreover,
the addition of catechol groups to GT will also enhance the adhe-
siveness of GT-DA/CS-based hydrogels due to the physical bonding
and chemical crosslinking between the catechol or polydopamine
group and the wounded tissue [15]. Additionally, the good adhe-
sion properties also allow the hydrogel to seal the wound, achiev-
ing a good hemostatic effect [16]. Therefore, grafting dopamine
onto GT will give the GT-DA/CS hydrogel good adhesiveness and
hemostatic and antioxidant properties, making it an excellent can-
didate for use in wound dressings.
In addition, researchers have found that the human body has
endogenous bioelectric systems. The surface of intact human skin
carries a more negative charge than do the deeper skin layers
[17]. However, when a defect or wound occurs in the skin, the dee-
per cells of the epidermis and the cells of the wound are positively
charged. The combination of a positively charged wound and neg-
atively charged surrounding intact skin creates what is called a
skin battery [18]. This bioelectric current facilitates wound healing
best when the wound tissue is moistened. The roles of conductive
materials in promoting wound healing have also been demon-
strated in previous studies [19–25]. Due to their excellent mechan-
ical, thermal and electronic properties, carbon nanotubes (CNTs)
have led the boom in nanotechnology research in the past decades
[26–28]. Meanwhile, CNTs-embedded scaffolds [29,30] have been
demonstrated to enhance electrical properties and facilitate signal
propagation, consequently improving cell-cell coupling [31].
515
However, the strong hydrophobic interaction between individual
CNTs makes it difficult for the original CNTs to disperse in water
[32]. Surface coating has been demonstrated to be an efficient
way to improve the CNTs dispersion and to reduce their cytotoxi-
city [33,34]. Dopamine coating is an easy and effective way to
modify the surface of CNTs [35], as dopamine will self-
polymerize in alkaline solution and form a layer of polydopamine
(PDA) coating on the surfaces of CNTs [31], which avoids the disad-
vantages of conventional oxidation modification strategies. Addi-
tionally, CNTs can strongly absorb near-infrared (NIR) energy and
efficiently convert it into thermal energy, and they also exhibit
good photothermal antibacterial effects, which will benefit the
healing of infected wounds [36,37]. Thus, developing CNT-PDA
and GT-DA/CS-based antibacterial adhesive antioxidative conduc-
tive hydrogels is highly desirable to heal infected wounds.
Here, chitosan, gelatin-grafted-dopamine and polydopamine-
coated CNTs were used as building blocks to engineer antibacterial
adhesive conductive GT-DA/CS/CNT composite hydrogels by the
self-polymerization of dopamine using a H2O2/HRP catalytic sys-
tem, and their good therapeutic effect in the treatment of infected
full-thickness defect wounds was also demonstrated. Chitosan was
added into the system to improve the hydrogels’ mechanical prop-
erties and compensate for the disadvantage of rapid degradation.
The use of the H2O2/HRP catalytic system not only reduced the bio-
logical safety problems observed in the traditional oxidative meth-
ods but also produced similar or comparatively better mechanical
properties than the traditional chemical methods. The swelling
ratio, degradation behaviors, morphology and rheology properties
of the engineered GT-DA/CS/CNT hydrogels were characterized.
The properties of tissue adhesiveness, hemostatic and antioxidant
activities, conductivity, photothermal effects and sustained drug
release were also studied. Additionally, to endow antibacterial
ability to the composite hydrogels, doxycycline was loaded into
the polymeric network, and the antimicrobial properties of the
GT-DA/CS/CNT/Doxy hydrogels were evaluated against Gram-(+)
Staphylococcus aureus (S. aureus) and Gram-(
) Escherichia coli
(E. coli). Furthermore, in vitro cytocompatibility and blood compat-
ibility testing of the GT-DA/CS/CNT hydrogels was also conducted.
Last, an infected mouse full-thickness skin defect wound model
was used to evaluate the therapeutic effects of these hydrogels.
These GT-DA/CS/CNT hydrogels’ multifunctional properties,
including antibacterial activity, adhesiveness and conductivity,
make them excellent candidates for the treatment of infected skin
wounds.
2. Materials and methods
2.1. Materials
Gelatin (Type A, gel strength  300 g Bloom, Sigma-Aldrich),
dopamine hydrochloride (DA, Sigma-Aldrich, 98%), 1-(3-dimethyla
minopropyl)-3-ethylcarbodiimide hydrochloride (EDC, J&K Chemi-
cal, >98.5%), N-Hydroxysuccinimide (NHS, Sigma-Aldrich, 98%),
carbon nanotube (CNTs, XFNANO, Materials Tech CO. Ltd.), 2,2-
Diphenyl-1-(2, 4,6-trinitrophenyl)-hydrazyl (DPPH, J&K Chemical,
97%), horseradish peroxidase (HRP, J&K Chemical, 98%), and doxy-
cycline hydrochloride (J&K Chemical, 98%) were used as received.
2.2. Synthesis of gelatin-grafted-dopamine (GT-DA)
Gelatin-grafted-dopamine was synthesized via ethyl-dimethyl-
amino-propylcarbodiimide (EDC) and N-hydroxy-succinimide
(NHS) coupling chemistry. First, 10 mL of phosphate buffered sal-
ine (PBS, pH 5.0) was used to dissolve the gelatin (0.5 g, type A
from porcine skin, Sigma-Aldrich) at 50 by stirring. Then,
516 Y. Liang et al. / Journal of Colloid and Interface Science 556 (2019) 514–528
dopamine hydrochloride (0.741 g, Sigma-Aldrich) was added to the
mixture, followed by the addition of EDC (0.749 g, Sigma-Aldrich)
and NHS (0.45 g, Sigma-Aldrich). The pH value of the solution
was maintained at approximately 5.0 for 12 h at 37 . The reaction
solution was then dialyzed in deionized water for 48 h prior to
lyophilization [38].
2.3. Synthesis of CNT-PDA
PDA-coated CNTs were prepared according to a previous report
[39], and the details of the specific operational steps are available
in the Supporting Information (SI).
2.4. Preparation of GT-DA/CS/CNT hydrogels
The hydrogels were prepared by the polymerization of GT-DA/
CS and CNT-PDA using H2O2/HRP as the initiator system. First,
GT-DA/CS (95:5 w/w) was dissolved in PBS to form a 10 wt% solu-
tion. CNT-PDA was dispersed in PBS to form 1 wt%, 2 wt%, and 4 wt
% CNT-PDA dispersion solutions, respectively. Then, 1 mL of the
CNT-PDA dispersion solutions was added to the 7.5 mL of GT-DA/
CS solution. After thorough stirring, 0.5 mL H2O2 (0.2 mol/mL)
and 1 mL HRP (0.25 mg/mL) were added to the GT-DA/CS/CNT mix-
tures. Then, the sol-gel transformation was completed within a
short period of time. Finally, hydrogels with a constant 10 wt%
GT-DA/CS and concentrations of CNT-PDA to GT-DA/CS increasing
from 0 wt% to 1 wt%, 2 wt% and 4 wt% were prepared for further
study.
2.5. Characterizations
UV–vis spectroscopy, nuclear magnetic resonance (1H NMR),
Fourier-transform infrared spectroscopy (FT-IR), transmission elec-
tron microscopy (TEM) and scanning electron microscopy (SEM)
were used to investigate the physical and chemical characteristics
of the copolymer and hydrogel samples. The specific operational
steps of the conductivity [40,41], swelling [42,43] and degradation
tests [44,45] are available in the SI.
2.6. Rheological test of the hydrogels
The rheological testing of these hydrogels was carried out using
a TA rheometer (DHR-2) [46]. The specific operational steps are
available in the SI.
2.7. Mechanical properties of GT-DA/CS/CNT hydrogels
Compression testing of the GT-DA/CS/CNT hydrogels was per-
formed according to our previous reports [42]. The details are
available in the SI.
2.8. Antioxidant ability of hydrogels
The antioxidant ability of these GT-DA/CS/CNT hydrogels was
evaluated using the stable 1,1-diphenyl-2-picrylhydrazyl (DPPH)
free radical [47]. The specific operational steps are available in
the SI.
2.9. Adhesive strength testing of the hydrogels
To evaluate the tissue-adhesive properties of these hydrogels, a
lap-shear test was performed by referring to previous reports with
some modifications [48,49]. The details of the test are available in
the SI.
2.10. In vivo hemostatic ability testing
According to previous research [50], for evaluation of the hemo-
static ability of the GT-DA/CS/CNT hydrogels, a mouse model with
a bleeding liver (Kunming mice, 25–30 g, female) was established.
The details are available in the SI. All animal experiments in this
work were carried out in accordance with current guidelines for
the care of laboratory animals and were approved by the proper
committee of Xi’an Jiaotong University.
2.11. Photothermal effect of the hydrogels
Examination of the photothermal effect of the hydrogels was
performed by the modification of a previously described protocol
[51], the details of the test are available in the SI.
2.12. In vitro and in vivo photothermal antibacterial activity of the
hydrogels
The in vitro photothermal antibacterial activity was tested
according to previously described methods [52]. The in vivo pho-
tothermal antibacterial activity was also examined by the modifi-
cation of previously described protocols [53,54]. The specific
operational steps are available in the SI.
2.13. In vitro sustained drug release behavior study
Doxycycline was used as the model drug to evaluate the sus-
tained drug release profiles of the GT-DA/CS/CNT hydrogels
[55,56]. For the purpose of further investigating the mechanism
of drug release from the hydrogels, the release behavior was fitted
using the Korsmeyer-Peppas equation [57]. The details are avail-
able in the SI.
2.14. Zone of inhibition testing of antibiotic-loaded GT-DA/CS/CNT
hydrogels
The antibacterial activity of doxycycline-loaded GT-DA/CS/
CNT2 hydrogels against S. aureus and E. coli was evaluated using
a zone of inhibition test [58,59]. The details of the specific opera-
tional steps are available in the SI.
2.15. Hemolytic testing of the hydrogels
Hemolysis testing was carried out by the modification of a pre-
viously described protocol [60]. The details are available in the SI.
2.16. Cytocompatibility testing of the hydrogels
The cytocompatibility was evaluated using a direct contact test
between the hydrogels and L929 cells according to previous
research [42,61]. The details are available in the SI.
2.17. In vivo wound healing evaluation with an infected full-thickness
skin defect model
To further evaluate the effect of GT-DA/CS/CNT hydrogels on
promoting wound repair, an infected full-thickness skin defect
model was established. Our animal experiments were approved
by the institutional review board of Xi’an Jiaotong University.
Female Kunming mice weighing 25–35 g and of 5–6 weeks of age
were used for the studies. The mice were randomly divided into
4 groups including TegadermTM film (control), GT-DA/CS/CNT0
hydrogel, GT-DA/CS/CNT2 hydrogel and GT-DA/CS/CNT2/Doxy
hydrogel. After anesthesia and shaving, 7 mm-diameter skin
wounds were created, and 50 lL S. aureus (108 CFU/mL) was used
 Y. Liang et al. / Journal of Colloid and Interface Science 556 (2019) 514–528
to establish infection at 48 h after implantation. Then, the control
wounds were treated with 50 lL of PBS and dressed with Tega-
dermTM film (3 M Health Care, USA), while the hydrogel group
wounds were treated with 50 lL of GT-DA/CS/CNT0, GT-DA/CS/
CNT2 or GT-DA/CS/CNT2/Doxy hydrogel. Tissues were collected
from each of the 5 mice in the 4 groups on the 3rd, 7th, and 14th
days. All samples were stored at
80 °C before analysis. The regen-
eration process of the wounds was assessed according to the
wound closure ratio [62]. For biochemical analysis, on the 3rd,
7th and 14th days, disc-shaped tissue samples were collected
(diameter = 1 cm), and the collagen amount was evaluated by esti-
mating the hydroxyproline content using a commercial kit (Jian-
cheng Bioengineering, China). All operations followed the
manufacturer’s instructions [50].
2.18. Histology and immunohistochemistry
Histological and immunohistochemistry examinations were
carried out to estimate the blood vessel regeneration and inflam-
mation during the wound healing process. TGF-b and CD31 were
chosen for the immunohistochemistry staining [63]. These details
are shown in the SI.
2.19. Statistical analysis
All experimental data from the studies were analyzed statisti-
cally, and the results were expressed as the mean ± standard devi-
ation (SD). Statistical differences (*P < 0.05) were determined using
one-way ANOVA followed by a Bonferroni post hoc test for multi-
ple comparisons using SPSS, version 24 (IBM).
3. Results and discussion
3.1. Preparation of GT-DA/CS/CNT hydrogel
Herein, we present antimicrobial, adhesive, antioxidative and
conductive hydrogels to treat infected skin defect wounds. These
designed hydrogels were based on gelatin-grafted-dopamine and
polydopamine-coated CNTs. Gelatin, a highly regarded polymer
that provides structural integrity and mimics the native composi-
tion of the ECM to modulate cell function in human skin, is suitable
for skin regeneration applications. Because of its good tissue adhe-
sion characteristics, dopamine was chosen and conjugated to gela-
tin by classical EDC/NHS chemistry to produce GT-DA and
endowed adhesive, hemostatic and antioxidative properties to
the gelatin (Fig. 1a). 1H NMR data (Fig. S1) demonstrated the suc-
cessful grafting of dopamine onto the backbone of gelatin, and
the mass ratio of dopamine in the GT-DA was calculated by UV–
vis spectroscopy to be 3%. The addition of chitosan can further
reduce the disadvantages of gelatin hydrogels, such as their
mechanical weakness and fast degradation rate. GT-DA/CS/CNT
hydrogels with different CS contents (to GT-DA/CS copolymer
weight) varying from 0 wt% to 2.5 wt%, 5 wt% and 10 wt% were
prepared to prove the importance of CS to the stability of GT-DA/
CS/CNT hydrogels. As shown in Fig. S2, GT-DA/CS/CNT hydrogels
containing 0 wt% or 2.5 wt% CS degraded in whole or in large part
within 1 day. However, hydrogels with 5 wt% and 10 wt% CS were
still intact. As the weight ratio of CS increased between 0, 2.5%, 5%
and 10%, the remaining weight of the hydrogel increased from
0 mg to 8.5 mg, 14.4 mg and 14.8 mg. Therefore, the content of
CS was determined to be 5 wt% because of the resulting proper vis-
cosity of the solution and suitable stability of the GT-DA/CS/CNT
hydrogel. On the other hand, CNTs were chosen as the conductive
component on account of excellent electronic, mechanical and
thermal properties. The coating of polydopamine onto the surfaces
517
of CNTs under an alkaline environment enhanced their dispersion
and bioactivity and, in addition, reduced the cytotoxicity of the
CNTs (Fig. 1b). The successful synthesis of CNT-PDA was confirmed
by FT-IR (Fig. S3) and TEM (Fig. S4). As shown in Fig. 1c, the GT-DA/
CS solution and CNT-PDA dispersion liquid were mixed and formed
into a hydrogel network through oxidative coupling of dopamine
by using H2O2/HRP as the initiator system. By changing the con-
tents of CNT-PDA and GT-DA/CS from 0 wt% to 1 wt%, 2 wt% and
4 wt%, a series of GT-DA/CS/CNT hydrogels were synthesized, and
the samples were named GT-DA/CS/CNT0, GT-DA/CS/CNT1, GT-
DA/CS/CNT2 and GT-DA/CS/CNT4, respectively. Fig. 1d shows the
original, bending, compressing, stretching and recovery shapes
and the proven flexible mechanical properties of the hydrogels.
Images of tissue adhesion and wound dressing also show the
potential applications of these hydrogels.
3.2. Physical and chemical properties of the hydrogels
The time of the cross point of the storage modulus (G0 ) and loss
modulus (G00 ) obtained by rheological testing is considered to be
the gelation time. The GT-DA/CS/CNT0 hydrogel took the longest
time of approximately 2 min to form a hydrogel. When the mass
percentage of CNT-PDA increased to 1 wt%, 2 wt% and 4 wt%, the
gelation time gradually decreased to approximately 60 s, 36 s and
17 s, indicating that a higher concentration of CNT-PDA in the
hydrogel network was beneficial to enhancing the efficiency of
gelation.
The most remarkable feature of hydrogels is the hydrophilic
structure, which enables them to retain substantial water in their
polymeric networks and facilitates their absorption of wound exu-
date. Additionally, swelling is an important parameter which con-
trols the release behavior of drugs from these three-dimensional
networks. The swelling ratios of these hydrogels over time were
recorded and are shown in Fig. 2a. Overall, all of the GT-DA/CS/
CNT hydrogels exhibited a water absorption ratio varying from
20% to 40% of their initial wet weights after more than 3 days of
swelling. The swelling ratio was dependent on the concentration
of CNT-PDA. Specifically, when the GT-DA/CS/CNT0 hydrogel was
swollen in PBS for 84 h, the lowest swelling ratio of 26.2% was
observed. As the weight ratio of CNT-PDA increased from 1% to
2% and 4% in the GT-DA/CS/CNT1, GT-DA/CS/CNT2 and GT-DA/CS/
CNT4 hydrogels, the water absorption changed from 32.5% to
41.2% and 46.3%, respectively.
A high crosslinking density leads to a slower degradation rate,
and by adjusting the concentration of CNT-PDA, it was also proven
that these hydrogels’ degradation rate is adjustable. The maximum
mass retention of 25.9% was observed for the GT-DA/CS/CNT4
hydrogel after 15 days. Upon decreasing the concentration of
CNT-PDA, the remaining weight ratio also decreased from 21.4%
to 17.1% and 12.6% for the GT-DA/CS/CNT2, GT-DA/CS/CNT1 and
GT-DA/CS/CNT0 hydrogels, respectively. In addition, it was noted
that the hydrogels in all groups degraded quickly in the first three
days, but then, the degradation rate slowed. In conclusion, the test
results of swelling and degradation showed that increasing the
concentration of CNT-PDA in the hydrogel enhanced the cross-
linking efficiency and stability.
In addition to the good water retention of hydrogels, the inter-
connected porous structure also enables hydrogels to be similar to
the extracellular matrix structure, which provides good support for
their biomedical applications. The SEM images of hydrogels
showed an interconnected microstructure with uniform pore sizes
(Fig. 2c). The pore diameters of the hydrogels became smaller with
increasing CNT-PDA concentrations before swelling (decreasing
from 89 lm to 78, 72 and 63 lm for the GT-DA/CS/CNT0, GT-DA/
CS/CNT1, GT-DA/CS/CNT2 and GT-DA/CS/CNT4 hydrogels, respec-
tively) (Fig. 2d–e), and they obviously increased after swelling,
518 Y. Liang et al. / Journal of Colloid and Interface Science 556 (2019) 514–528

Fig. 1. Schematic representation of GT-DA/CS/CNT hydrogel preparation. Synthesis scheme of GT-DA polymer (a), CNT-PDA (b), and GT-DA/CS/CNT hydrogels (c); (d) Original,
bending, compressing and stretching shape representations and the application as a wound dressing. Scale bar: 5 mm.

with the mean value still showing dependence on the CNT-PDA
concentration.
Rheological properties testing of these hydrogels was carried
out to evaluate the effect of the CNT-PDA content on the mechan-
ical strength. The G0 and G00 of these hydrogels were recorded over
time. As shown in Fig. 2f, G0 became higher than G00 after a very
short time, indicating gelation through catechol group self-
polymerization. Specifically, the G0 of these hydrogels decreased
from 2900.8 Pa to 2519.6 Pa and 2016.1 Pa with decreasing CNT-
PDA weight ratios from 4 wt% to 2 wt% and 1 wt% and then
decreased to 981.5 Pa when the weight ratio of CNT-PDA became
0 wt%. These results demonstrated that the 4 wt% CNT-PDA weight
ratio led to a higher crosslinking density and more stable polymer
networks.
good elasticity under compression. Specifically, all of the GT-DA/
CS/CNT hydrogels could withstand high-level (80%) compression
without any observable damage or plastic deformation. The testing
also showed that incorporating CNT-PDA into the GT-DA/CS/CNT
hydrogels effectively enhanced the mechanical strength and that
hydrogels incorporating higher CNT-PDA concentrations also dis-
played higher compression stress (increasing from 68.0 to 78.9,
98.8, and 115.9 kPa at 80% strain with increasing CNT-PDA con-
tents from 0 wt% to 1 wt%, 2 wt%, and 4 wt%, respectively) at the
same strain. Additionally, after 20 cycles of compression-recovery
tests at 60% strain, the stress-strain diagram of the compression-
recovery of the hydrogels presented loops similar to the original
hydrogels (Fig. 2h–k). In addition, the recovered hydrogels still pre-
sented good shape, elasticity and high compressive strength,
exhibiting the good shape-recovery abilities of these hydrogels.

3.3. Mechanical properties and shape recovery of the GT-DA/CS/CNT 3.4. Tissue adhesion and in vivo hemostatic ability of GT-DA/CS/CNT
hydrogels hydrogels

Compression stress-strain testing was performed on the GT-DA/ Compared with traditional wound dressings, seamless wound
CS/CNT hydrogels. As shown in Fig. 2g, these hydrogels exhibited treatment shows great advantages [64]. A lap-shear measurement
 Y. Liang et al. / Journal of Colloid and Interface Science 556 (2019) 514–528 519

Fig. 2. Characterizations of the GT-DA/CS/CNT hydrogels. (a) Swelling properties; (b) Degradation profiles, with the data shown as the mean ± SD (n = 3); (c) SEM images,
scale bar: 50 lm; (d) Pore diameter distributions of different hydrogels before swelling and (e) after swelling, unit: lm; (f) Rheological behavior of the hydrogels; (g) The
uniaxial compression stress-strain diagram of the hydrogels; the stress–strain cycling diagram of hydrogels (h) GT-DA/CS/CNT0, (i) GT-DA/CS/CNT1, (j) GT-DA/CS/CNT2, and
(k) GT-DA/CS/CNT4 at the compression strain of 60%.

was performed to estimate the adhesion performance of these
hydrogels relative to skin (Fig. 3a, b) [49]. The test results showed
that the shear adhesive strength increased from 3.9 ± 1.2 kPa (GT-
DA/CS/CNT0) to 4.9 ± 1.1 kPa (GT-DA/CS/CNT1), 5.4 ± 0.6 kPa (GT-
DA/CS/CNT2) and 6.5 ± 0.9 kPa (GT-DA/CS/CNT4) (Fig. 3a), indicat-
ing that increasing the CNT-PDA concentration from 1 wt% to 2 wt
% and 4 wt% enhanced the adhesive properties of the resultant
hydrogels. As a skin wound-healing dressing, hydrogels will
directly contact the external environment. Therefore, after success-
ful preparation, samples were placed at 37 °C for 1 h before testing,
and the test results exhibited that the shear adhesive strength was
greatly enhanced to approximately 20–30 kPa (Fig. 3b). These
520 Y. Liang et al. / Journal of Colloid and Interface Science 556 (2019) 514–528

Fig. 3. Adhesive properties of different hydrogels (a) and adhesive strengths of samples after placement in air for 1 h before test (b); (c) Hemostatic ability of GT-DA/CS/CNT2
hydrogels; (d) Conductivity of the hydrogels; (e) DPPH scavenging rate of different concentrations of GT-DA/CS/CNT2 hydrogels; (f) UV–vis spectra of DPPH scavenging; (g)
Doxycycline release curves; (h) The release profiles of GT-DA/CS/CNT hydrogels were well-fitted by the Korsmeyer-Peppas equation; (i) Statistics of the diameter of the zone
of inhibition, with the data shown as the means ± SD (n = 3).

results exhibited the potential for the application of GT-DA/CS/CNT
hydrogels as adhesive wound dressings for wound closure and
healing.
Among the four stages of wound repair, hemostasis is the first
stage, which also shows its importance to the whole repair process.
Hydrogels with adhesion properties can seal the wound instantly
and show good hemostatic performance [65]. A mouse hepatic
bloodletting model was used to estimate the hemostatic effect of
these hydrogels, and the GT-DA/CS/CNT2 hydrogel was selected
as a representative. As shown in Fig. 3c, nearly 700 mg blood trick-
led from the hepar of mice in the untreated group. In contrast, the
GT-DA/CS/CNT2 hydrogel group exhibited as low as a quarter
(170 mg) of the bleeding of the control group. A significant differ-
ence in bloodshed amount between the control and GT-DA/CS/
CNT2 hydrogel groups was calculated with P < 0.01, which proved
the excellent in vivo hemostatic ability of the GT-DA/CS/CNT
hydrogels.
3.5. Conductivity and antioxidant ability of the GT-DA/CS/CNT
hydrogels
Conductive materials have been shown to promote wound heal-
ing, and loading carbon nanotubes into various synthetic polymer
matrices can markedly improve the electrical conductivity [38,66].
The conductivities of these GT-DA/CS/CNT hydrogels were tested
[67]. The GT-DA/CS/CNT0 hydrogel possessed the lowest conduc-
tivity of 2.5  10
2 S/m. When CNT-PDA was added, the conductiv-
ities improved from 6.2  10
2 to 6.7  10
2 and 7.2  10
2 S/m
for the GT-DA/CS/CNT1, GT-DA/CS/CNT2 and GT-DA/CS/CNT4
hydrogels, respectively (Fig. 3d), which would be beneficial for
mediating electrical signals between tissues and accelerating the
rate of wound repair by improving blood flow, enhancing migra-
tion and reducing edema [18].
Free radicals play a vital role in all stages of wound healing.
Topical applications of free radical-scavenging materials have been
 Y. Liang et al. / Journal of Colloid and Interface Science 556 (2019) 514–528
proven to accelerate wound repair [68,69]. The scavenging effi-
ciency for DPPH was used to estimate the antioxidant activity of
these hydrogels. It is shown in Fig. 3e that 3 mg/mL copolymer
scavenged nearly 86.5% of free radicals, and when the hydrogel
concentration increased to 5 mg/mL, almost all free radicals were
scavenged. In conclusion, the antioxidant activity of GT-DA/CS/
CNT hydrogels make them a competitive candidate for use as skin
wound repair dressings.
3.6. Drug release capability of the hydrogels
In clinics, antibiotics are the most common and useful treat-
ment for infection. Hence, as a widely used ‘second-generation’
tetracycline, doxycycline has been selected as a model drug to
endow these hydrogels with antibacterial properties, and the
cumulative release of doxycycline from these hydrogels over time
was recorded. As shown in Fig. 3g, in the beginning, the doxycy-
cline was released relatively quickly, and almost half of the drug
was released within the first 20 h. Then, although the rate of doxy-
cycline release slowed, the drug release course lasted almost 100 h,
and finally, nearly 80% of the doxycycline was released into PBS at
pH 7.4 from the four hydrogel groups. These doxycycline-release
behaviors of the GT-DA/CA/CNT hydrogels demonstrated their
potential for use as drug release carriers. To further investigate
the doxycycline-release behavior of GT-DA/CS/CNT hydrogels, we
found that the Korsmeyer-Peppas equation could be used to fit
the release curves with high correlation (r2 > 0.99).
n
Mt =M1 ¼ k  t
In this equation, n stands for the diffusion exponent. If n is
lower than 0.45 (for cylindrical samples), the method of drug
release can be defined as Fickian diffusion. As shown in Fig. 4h,
the n values of these hydrogels were all less than 0.5, indicating
that the release behavior of doxycycline from GT-DA/CA/CNT
hydrogels is diffusion.
To further confirm the effect of the released doxycycline from
the GT-DA/CS/CNT2/Doxy hydrogels, zone of inhibition testing
was performed with E. coli and S. aureus as the representative bac-
teria (Fig. S5). The PBS control and drug-free GT-DA/CS/CNT2
hydrogel group exhibited no zones of inhibition during the entire
process of the test. In contrast, the whole-release and bacterial
inactivation processes of antibiotics from the GT-DA/CS/CNT2/
Doxy hydrogels lasted 6 days and 9 days for E. coli and S. aureus,
respectively. The diameters of the zones of inhibition for S. aureus
and E. coli also decreased from 3.0 cm to 1.6 cm and 2.5 cm to
1.5 cm, respectively. This result was mainly because the amount
of doxycycline released from those hydrogels gradually decreased
(Fig. 3i).
3.7. Photothermal property and photothermal antibacterial activity
It is well known that CNTs can generate heat by light absorption
due to their strong NIR absorption. The photothermal ability of
these GT-DA/CS/CNT hydrogels was tested. The curves of DT as a
function of the NIR irradiation time for all four GT-DA/CS/CNT
hydrogels were recorded (Fig. 4a). The GT-DA/CS/CNT0 hydrogel
showed about a 10 °C increase in temperature after 10 min NIR
lighting (1.0 W/cm2) because quinones from oxidized catechol
groups possess some photothermal ability [70]. When CNT was
added to the hydrogel, the DT was obviously increased, and as
the CNT-PDA concentration increased from 1 wt% to 2 wt% and
4 wt%, the temperature increment increased from 17.2 °C to
21.3 °C and 23.3 °C, respectively, after 10 min irradiation. The core
region possessed the highest temperatures, ranging from 35 °C to
50 °C, as shown in Fig. S6. All of the results confirmed the excellent
521
photothermal effect of the CNT-PDA contained in GT-DA/CS/CNT
hydrogels.
Although antibiotics can play a good antibacterial role in clinics,
their abuse has become increasingly serious. Researchers have
found that temperatures above 50 degrees can kill bacteria. The
photothermal antibacterial efficiency of the GT-DA/CS/CNT2
(DT = 21.3 °C) hydrogels was estimated, and the GT-DA/CS/CNT0
hydrogel and PBS were used as controls [71]. As shown in the rep-
resentative photos (Fig. 4b), the GT-DA/CS/CNT0 and PBS groups
did not show any macroscopic change in the number of bacteria
under different irradiation times. However, the survival S. aureus
and E. coli obviously decreased with 5 min irradiation, and the bac-
teria were completely invisible with 10 min irradiation for the GT-
DA/CS/CNT2 hydrogels. The quantitative data in Fig. 4d and e also
show consistency with Fig. 4b. When CNT-PDA was added into the
hydrogel, only 3 min of NIR irradiation to the GT-DA/CS/CNT2
hydrogel caused a significant decrease (P < 0.05) in the bacteria
survival ratio, and only 5.9% of S. aureus and 2.1% of E. coli was still
alive. Furthermore, surviving S. aureus and E. coli were almost
invisible as the irradiation time was prolonged to 5 min. By
increasing the irradiation time to 10 min, GT-DA/CS/CNT2 group
exhibited 100% bacteria killing for both S. aureus and E. coli
(Fig. 4d and e). In addition, only 75% S. aureus and 62% E. coli were
still alive after 10 min irradiation to the GT-DA/CS/CNT0 hydrogels,
which benefited from the photothermal effect of the PDA con-
tained in the CNT-PDA. In conclusion, the good in vitro near-
infrared enhanced photothermal antibacterial capability of these
GT-DA/CS/CNT hydrogels was verified, which also increased the
ð1Þ superior performance of these hydrogels in the face of complex
infections.
At present, gram-positive bacteria, especially S. aureus, are the
main cause of wound infection. An in vivo antibacterial test was
further performed to demonstrate the photothermal antibacterial
activity of these hydrogels. An infected mouse skin defect model
was used to carry out this test. Representative photos of the
in vivo antibacterial activity toward S. aureus (Fig. 4c) show a
macroscopic decrease in the number of colonies between the GT-
DA/CS/CNT2 + NIR hydrogel group and the other three groups
(PBS control group, PBS + NIR group and hydrogel GT-DA/CS/
CNT2 group) after the application of 10 min NIR irradiation. The
quantitative data in Fig. 4f also show the obviously lower (3.1%,
P < 0.01) bacterial survival ratios of GT-DA/CS/CNT2 + NIR hydro-
gels when compared with the other three groups. Additionally,
the three control groups showed no significant differences, show-
ing that the lower bacterial survival mainly originated from the
photothermal ability of CNTs. In conclusion, all of the test results
demonstrated the great in vitro and in vivo photothermal-induced
antibacterial activity of GT-DA/CS/CNT hydrogels.
3.8. Cytocompatibility and hemocompatibility evaluation of the
hydrogels
The basic property requirement of biomaterials is good compat-
ibility. Therefore, a direct contact test with L929 fibroblast cells
was performed to evaluate the hydrogels’ in vitro cytotoxicity. As
shown in Fig. 4g, the cell viability exhibited no obvious differences
among all the groups on the 1st and 3rd day, demonstrating the
nontoxicity of the GT-DA/CS/CNT hydrogels. After 5 days of co-
incubation, although all of the hydrogel groups showed lower cell
proliferation than that of the TCP on average, only the GT-DA/CS/
CNT4 hydrogel exhibited a significant difference (P < 0.05) relative
to the TCP. The images in Fig. 4h also reveal results consistent with
those of the quantitative experiments.
The hemocompatibility of the GT-DA/CS/CNT hydrogels was
estimated. The picture in Fig. 4i shows obvious differences in color
among the four hydrogel groups, PBS group and Triton X-100
522 Y. Liang et al. / Journal of Colloid and Interface Science 556 (2019) 514–528

Fig. 4. (a) DT-NIR lighting time diagrams of hydrogels with 1.0 W/cm2 light intensity; (b) NIR lighting-induced in vitro antibacterial activity of hydrogels for in vitro
antibacterial activity, with 0, 1, 3, 5 and 10 representing the different irradiation times (min); (c) Photographs of the in vivo antibacterial activity, with 1–5 indicating the
blank, PBS treated, PBS + NIR irradiation, GT-DA/CS/CNT2 and GT-DA/CS/CNT2 + NIR groups, respectively. Bacterial survival ratios of (d) S. aureus and (e) E. coli; (f) Bacterial
survival ratios of S. aureus in vivo in the antibacterial activity test, with the data shown as the means ± SD (n = 3) (g) Cell compatibility evaluation, *P < 0.05; (h) LIVE/DEAD
staining of L929 cells after 5 days of co-culture with the hydrogels. Scale bar: 200 lm, with the data shown as means ± SD (n = 5); (i) Photographs from the hemolytic assay;
(j) Quantitative data of the hemolytic ratio, with the data shown as means ± SD (n = 3).

group. All of the hydrogel groups were pale yellow, which was
comparable to PBS and obviously different from the Triton X-100
group (red due to complete hemolysis). When the hemolysis ratio
of the positive group was set to 100%, all of the hydrogel groups
exhibited hemolytic ratios of less than 4%, even though the weight
ratio of CNT-PDA was increased to 4 mg/mL in the hydrogels
(Fig. 4j). The GT-DA/CS/CNT0 and GT-DA/CS/CNT1 hydrogels even
exhibited less than 1.5% hemolysis. The GT-DA/CS/CNT2 hydrogel
was chosen for further testing due to its balanced cell proliferation
and mechanical and photothermal properties.
3.9. In vivo wound healing evaluation with an infected full-thickness
skin defect model
An infected full-thickness mouse skin defect model was per-
formed to evaluate the actual effect of GT-DA/CS/CNT hydrogels
in promoting infected wound healing. The GT-DA/CS/CNT2 hydro-
gel was selected as the representative, and a doxycycline
antibiotic-encapsulated GT-DA/CS/CNT2/Doxy hydrogel was used
in another experimental group. The commercially available Tega-
dermTM film was chosen as a control. It can be seen from Fig. 5a that
 Y. Liang et al. / Journal of Colloid and Interface Science 556 (2019) 514–528 523

Fig. 5. (a) Pictures of wounds at 3 days, 7 days and 14 days for TegadermTM film, GT-DA/CS/CNT0, GT-DA/CS/CNT2 and GT-DA/CS/CNT2/Doxy hydrogel; (b) Wound closure for
each group. *P < 0.05; (c) Hydroxyproline content in the new skin tissue. *P < 0.05; (d) Pictures of granulation tissue on the 7th day (green arrows). Scale bar: 200 lm. (e)
Granulation tissue thicknesses for different groups on the 7th day, with the data shown as the means ± SD (n = 5). (For interpretation of the references to color in this figure
legend, the reader is referred to the web version of this article.)

after treatment for 3 days, compared with the TegadermTM film
group, an obviously higher degree of wound closure was observed
for the GT-DA/CS/CNT0, GT-DA/CS/CNT2 and GT-DA/CS/CNT2/
Doxy hydrogels (P < 0.05), revealing the better wound healing
effect of the GT-DA/CS/CNT hydrogels (Fig. 5b). The GT-DA/CS/
CNT2/Doxy hydrogel also exhibited an obviously reduced wound
area (P < 0.05) compared to that of the GT-DA/CS/CNT0 and GT-
DA/CS/CNT2 hydrogels, demonstrating the positive effect of antibi-
otics on the repair of infected wounds. After 7 days and 14 days, all
of the hydrogel groups still exhibited obviously better (P < 0.05)
repair effects compared to TegadermTM film. Additionally, on the
14th day, the wound in the TegadermTM film group was still macro-
scopically unclosed, but the wounds covered with hydrogels were
nearly all closed and even showed smooth new epidermal tissue.
Significantly better wound closure occurred in the GT-DA/CS/
CNT2/Doxy group in comparison with those in the GT-DA/CS/
CNT0 and GT-DA/CS/CNT2 groups (P < 0.05), which benefited from
the addition of antibiotics. It is worth noting that the GT-DA/CS/
CNT2 hydrogel also showed a significantly reduced wound area
compared with the GT-DA/CS/CNT0 hydrogel on the 3rd day and
7th day, proving the promoting effect of CNT-PDA in wound clo-
sure. In conclusion, the wound healing results indicated that GT-
DA/CS/CNT hydrogels with adhesive and hemostatic properties
exhibited a good wound healing effect compared with the Tega-
dermTM film group, especially for hydrogels containing CNT-PDA
and antibiotics.
As an indispensable process in skin wound healing, collagen
deposition is an important indicator in the evaluation of healing
effects. Therefore, the hydroxyproline content in the new skin tis-
sue was tested as representative of collagen deposition to estimate
the repair effect. As shown in Fig. 5c, the collagen deposition levels
exhibited a great increase from the 3rd day to the 7th day and then
a slight elevation on the 14th day. In addition, all hydrogel groups
showed higher collagen levels on the 3rd and 7th day of the repair
process compared to the TegadermTM film group. In particular, the
GT-DA/CS/CNT2/Doxy and GT-DA/CS/CNT2 groups exhibited
higher collagen depositions than did the GT-DA/CS/CNT0 group
(p < 0.05) on the 7th day.
Approximately four days after trauma occurs, granulation tissue
begins to form in the wound, and then macrophages, fibroblasts
524 Y. Liang et al. / Journal of Colloid and Interface Science 556 (2019) 514–528

and blood vessels begin to appear in the granulation tissue, provid-
ing growth factors, new extracellular matrix and oxygen and nutri-
ents to promote wound healing [72]. Therefore, the thickness of
granulation tissue can reflect the quality of wound repair. As
shown in Fig. 5d and e, three hydrogel groups exhibited thicker
granulation tissue (P < 0.05) than the control group. Additionally,
the GT-DA/CS/CNT2/Doxy hydrogel produced thicker granulation
tissue than did the GT-DA/CS/CNT0 and GT-DA/CS/CNT2 hydrogels,
indicating its better wound healing effect.
3.10. Histomorphological evaluation
The regenerated skin tissue of the four groups during wound
healing process was further assessed through histological analysis
[73]. As shown in Fig. 6, there were fewer inflammatory cells in
wound sites of the three hydrogel groups than in those of the Tega-
dermTM film group on day 3 (Fig. 6a), especially for the antibiotic-
loaded GT-DA/CS/CNT/Doxy hydrogel group. On the day 7, there
were fewer inflammatory cells in all the groups. In addition, all
the hydrogel groups exhibited a significantly thicker epidermis
than did the TegadermTM film group on day 7 (P < 0.05), especially
the GT-DA/CS/CNT2 and GT-DA/CS/CNT2/Doxy groups (Fig. 6b).
Furthermore, the GT-DA/CS/CNT2 group showed a thicker epider-
mis compared with that of the GT-DA/CS/CNT0 group due to the
addition of CNT-PDA, and the epidermal regeneration for the GT-
DA/CS/CNT2/Doxy group was also better than that for the GT-DA/
CS/CNT2 group due to the addition of antibiotics. Blood vessels
entered the wound site approximately 4 days after injury, and they
could deliver oxygen and nutrients to support cellular activity [74].
The number of new blood vessels is also a vital indicator for esti-

Fig. 6. (a) Histomorphological measurements of wound healing for the TegadermTM film, GT-DA/CS/CNT0, GT-DA/CS/CNT2 and GT-DA/CS/CNT2/Doxy hydrogel groups after
3 days, 7 days and 14 days. Scale bar: 100 lm. (b) The regenerated epidermis thickness after 7 days; (c) Blood vessel regeneration after 7 days and 14 days; (d) Number of hair
follicles after 14 days, with the data shown as the means ± SD (n = 5).
 Y. Liang et al. / Journal of Colloid and Interface Science 556 (2019) 514–528 525

mating the repair effect. Fig. 6c exhibits an increasing trend of
regenerated blood vessels in the defect skin wounds from the 7th
day to the 14th day after treatment. Statistics of the regenerated
blood vessels confirmed that the wounds in the GT-DA/CS/CNT2
and GT-DA/CS/CNT2/Doxy hydrogel groups possessed improved
vascularization compared to that in the TegadermTM films group
and GT-DA/CS/CNT0 group (P < 0.05). On day 14, almost complete
regeneration of dermis tissue with appendages was observed in the
wounds treated with GT-DA/CS/CNT0, GT-DA/CS/CNT2 and GT-DA/
CS/CNT2/Doxy hydrogels (Fig. 6a) [70], indicating their better
regeneration. However, the wound in the TegadermTM film group
did not exhibit complete re-epithelialization, and even signifi-
cantly fewer hair follicles were observed in the dermis tissue com-
pared with that in the hydrogel groups (Fig. 6d, P < 0.05). The
in vivo infected wound healing test results indicated that all of
the hydrogel groups favored tissue regeneration and physiological
structure remodeling. CNT-PDA-containing hydrogels can effec-
tively reconstruct dermis and epidermis tissue by reducing infec-
tion, transmitting electrical signals and stimulating cells between
the wound through antibacterial activity, conductivity and opti-
mized physicochemical properties. In conclusion, all of these prop-
erties make GT-DA/CS/CNT hydrogels highly promising
multifunctional biomaterials for potential wound healing
applications.
3.11. TGF-b and CD31 expression during wound healing
Transforming growth factor beta (TGF-b) shows multiple effects
on skin wound healing [75]. TGF-b also plays a vital role in promot-
ing further inflammation [76]. It can be seen from Fig. 7a and c that
the expression of TGF-b showed a general trend of gradual decreas-
ing over time during the whole repair process, which also con-
firmed the smooth progress of the repair process [77]. Further
analysis showed that all of the hydrogel groups revealed overall
lower TGF-b expressions than that in the control group on the third
day after treatment, confirming the better effect of GT-DA/CS/CNT

Fig. 7. Images of immunofluorescence labeling of skin wound tissues on the 3rd day, 7th day and 14th day with (a) TGF-b and (b) CD31 on day 7, Scale bar: 50 lm.
Quantitative data of the area coverage by (c) TGF-b and (d) CD31, respectively. Data for the TegadermTM films on the 3rd day for TGF-b and the 7th day for CD31 were set as
100%. *P < 0.05, with the data shown as the means ± SD (n = 3).
526 Y. Liang et al. / Journal of Colloid and Interface Science 556 (2019) 514–528
hydrogels in reducing infection. Due to the addition of the antibi-
otic doxycycline, the GT-DA/CS/CNT2/Doxy hydrogel naturally
resulted in lower TGF-b than did the other three groups. Moreover,
the GT-DA/CS/CNT2 group also showed better results than those of
the GT-DA/CS/CNT0 group, demonstrating the promoting effect of
the addition of CNTs on wound healing. The same patterns were
also observed on the 7th and 14th day, proving that the effective
control of inflammation in the early stage of wound repair has a
positive effect on the whole repair process. CD31 was chosen to
estimate the regeneration of vessels because it is expressed during
early vascular reconstruction. As shown in Fig. 7b and d, wounds
treated with GT-DA/CS/CNT0, GT-DA/CS/CNT2 and GT-DA/CS/
CNT2/Doxy hydrogels exhibited higher expressions of CD31 than
did those treated with TegadermTM films after 7 days, confirming
that the hydrogels, especially those which contained CNTs and
antibiotics, promoted angiogenesis. In conclusion, according to
the results of immunofluorescence analysis, the hydrogel groups
overall showed a better effect in promoting wound healing, prob-
ably due to the reduced expression of TGF-b and the upregulated
expression of CD31.
4. Conclusions
We present a series of antibacterial adhesive antioxidant and
conductive GT-DA/CS/CNT composite hydrogels produced through
the oxidative coupling of catechol groups between gelatin-grafted-
dopamine and polydopamine-coated CNTs. The addition of the
antibiotic doxycycline endowed the hydrogel with antimicrobial
activity, and these multifunctional hydrogels showed great poten-
tial for promoting the repair of infected wounds [78,79]. The poros-
ity, rheology, mechanical properties, conductivity, swelling and
degradability of these hydrogels were demonstrated to be related
to the concentration of CNT-PDA. CNT-PDA endowed better pho-
tothermal and broad-spectrum photothermal antimicrobial activi-
ties, and dopamine endowed tissue-adhesive, hemostatic and
better antioxidative properties, to these hydrogels compared to
our previous research, which was verified [64]. Additionally, the
better drug release [80] and zone of inhibition results of the
doxycycline-encapsulated hydrogels exhibited sustained drug
release profiles and good antibacterial properties. The examination
of hemolysis and co-culturing with L929 cells confirmed the good
in vitro biocompatibility of these hydrogels. The better promoting
effects of these hydrogels in infected skin wound repair were
demonstrated by means of wound closure [81], collagen metabo-
lism, granulation tissue thickness [82], epidermis regeneration,
hair follicles and blood vessels and immunofluorescence staining
of TGF-b and CD31. Further evaluations are still in progress in
our laboratory. In conclusion, the antibacterial, adhesive, antiox-
idative, and conductive GT-DA/CS/CNT hydrogels are competitive
candidates for use in treatment of infected skin defect wounds.
Declaration of Competing Interest
The authors declare no competing financial interest.
Acknowledgement
This work was jointly supported by the National Natural
Science Foundation of China (grant number: 51673155), the State
Key Laboratory for Mechanical Behavior of Materials, the Funda-
mental Research Funds for the Central Universities, the World-
Class Universities (Disciplines), the Characteristic Development
Guidance Funds for the Central Universities, the Natural Science
Foundation of Shaanxi Province (Nos. 2018JM5026, and 2019TD-
020), and the Opening Project of Key Laboratory of Shaanxi Pro-
vince for Craniofacial Precision Medicine Research, College of
Stomatology, Xi’an Jiaotong University (No. 2019LHM-KFKT008).
Appendix A. Supplementary material
Supplementary data to this article can be found online at
https://doi.org/10.1016/j.jcis.2019.08.083.
References
[1] G. Han, R. Ceilley, Chronic wound healing: a review of current management
and treatments, Adv. Ther. 34 (3) (2017) 599–610.
[2] H. Liu, C. Wang, C. Li, Y. Qin, Z. Wang, F. Yang, Z. Li, J. Wang, A functional
chitosan-based hydrogel as a wound dressing and drug delivery system in the
treatment of wound healing, RSC Adv. 8 (14) (2018) 7533–7549.
[3] M.L. Mangoni, A.M. McDermott, M. Zasloff, Antimicrobial peptides and wound
healing: biological and therapeutic considerations, Exp. Dermatol. 25 (3)
(2016) 167–173.
[4] S. Dhivya, V.V. Padma, E. Santhini, Wound dressings–a review, BioMedicine 5
(4) (2015) 22.
[5] N. Zhao, J. Coyne, M. Xu, X. Zhang, A. Suzuki, P. Shi, J. Lai, G.-H. Fong, N. Xiong,
Y. Wang, Assembly of bifunctional aptamer-fibrinogen macromer for VEGF
delivery and skin wound healing, Chem. Mater. 31 (3) (2019) 1006–1015.
[6] J. Zhao, B. Guo, P.X. Ma, Injectable alginate microsphere/PLGA–PEG–PLGA
composite hydrogels for sustained drug release, RSC Adv. 4 (34) (2014) 17736–
17742.
[7] H. Ueno, F. Nakamura, M. Murakami, M. Okumura, T. Kadosawa, T. Fujinaga,
Evaluation effects of chitosan for the extracellular matrix production by
fibroblasts and the growth factors production by macrophages, Biomaterials
22 (15) (2001) 2125–2130.
[8] Y. Huang, S. Onyeri, M. Siewe, A. Moshfeghian, S.V. Madihally, In vitro
characterization of chitosan–gelatin scaffolds for tissue engineering,
Biomaterials 26 (36) (2005) 7616–7627.
[9] L. Fan, H. Yang, J. Yang, M. Peng, J. Hu, Preparation and characterization of
chitosan/gelatin/PVA hydrogel for wound dressings, Carbohyd. Polym. 146
(2016) 427–434.
[10] R. Moseley, J.E. Stewart, P. Stephens, R.J. Waddington, D.W. Thomas,
Extracellular matrix metabolites as potential biomarkers of disease activity
in wound fluid: lessons learned from other inflammatory diseases?, Brit J.
Dermatol. 150 (3) (2004) 401–413.
[11] C. Dunnill, T. Patton, J. Brennan, J. Barrett, M. Dryden, J. Cooke, D. Leaper, N.T.
Georgopoulos, Reactive oxygen species (ROS) and wound healing: the
functional role of ROS and emerging ROS-modulating technologies for
augmentation of the healing process, Int. Wound J. 14 (1) (2017) 89–96.
[12] W. Chen, X. Shen, Y. Hu, K. Xu, Q. Ran, Y. Yu, L. Dai, Z. Yuan, L. Huang, T. Shen, K.
Cai, Surface functionalization of titanium implants with chitosan-catechol
conjugate for suppression of ROS-induced cells damage and improvement of
osteogenesis, Biomaterials 114 (2017) 82–96.
[13] A.I. Neto, A.C. Cibrao, C.R. Correia, R.R. Carvalho, G.M. Luz, G.G. Ferrer, G.
Botelho, C. Picart, N.M. Alves, J.F. Mano, Nanostructured polymeric coatings
based on chitosan and dopamine-modified hyaluronic acid for biomedical
applications, Small 10 (12) (2014) 2459–2469.
[14] S. Hong, K. Yang, B. Kang, C. Lee, I.T. Song, E. Byun, K.I. Park, S.W. Cho, H. Lee,
Hyaluronic acid catechol: a biopolymer exhibiting a pH-dependent adhesive or
cohesive property for human neural stem cell engineering, Adv. Funct. Mater.
23 (14) (2013) 1774–1780.
[15] J.H. Ryu, S. Hong, H. Lee, Bio-inspired adhesive catechol-conjugated chitosan
for biomedical applications: a mini review, Acta Biomater. 27 (2015) 101–115.
[16] L. Han, L. Yan, M. Wang, K. Wang, L. Fang, J. Zhou, J. Fang, F. Ren, X. Lu,
Transparent, adhesive, and conductive hydrogel for soft bioelectronics based
on light-transmitting polydopamine-doped polypyrrole nanofibrils, Chem.
Mater. 30 (16) (2018) 5561–5572.
[17] I. Foulds, A.J. Barker, Human skin battery potentials and their possible role in
wound healing, Brit. J. Dermatol. 109 (5) (1983) 515–522.
[18] L.C. Kloth, J.M. McCulloch, Promotion of wound healing with electrical
stimulation, Adv. Wound Care 9 (1996) 42–45.
[19] B. Guo, A. Finne-Wistrand, A.-C. Albertsson, Facile synthesis of degradable and
electrically conductive polysaccharide hydrogels, Biomacromolecules 12 (7)
(2011) 2601–2609.
[20] B. Guo, L. Glavas, A.-C. Albertsson, Biodegradable and electrically conducting
polymers for biomedical applications, Prog. Polym. Sci. 38 (9) (2013) 1263–
1286.
[21] B. Guo, P.X. Ma, Conducting Polymers for Tissue Engineering,
Biomacromolecules 19 (6) (2018) 1764–1782.
[22] L. Wang, Y. Wu, B. Guo, P.X. Ma, Nanofiber yarn/hydrogel core–shell scaffolds
mimicking native skeletal muscle tissue for guiding 3D myoblast alignment,
elongation, and differentiation, ACS Nano 9 (9) (2015) 9167–9179.
[23] Y. Wu, L. Wang, B. Guo, P.X. Ma, Interwoven aligned conductive nanofiber
yarn/hydrogel composite scaffolds for engineered 3D cardiac anisotropy, ACS
Nano 11 (6) (2017) 5646–5659.
[24] T. Hu, Y. Wu, X. Zhao, L. Wang, L. Bi, P.X. Ma, B. Guo, Micropatterned,
electroactive, and biodegradable poly(glycerol sebacate)-aniline trimer
elastomer for cardiac tissue engineering, Chem. Eng. J. 366 (2019) 208–222.
 Y. Liang et al. / Journal of Colloid and Interface Science 556 (2019) 514–528
[25] Y. Wu, L. Wang, B. Guo, Y. Shao, P.X. Ma, Electroactive biodegradable
polyurethane significantly enhanced Schwann cells myelin gene expression
and neurotrophin secretion for peripheral nerve tissue engineering,
Biomaterials 87 (2016) 18–31.
[26] L. Yu, C. Shearer, J. Shapter, Recent development of carbon nanotube
transparent conductive films, Chem. Rev. 116 (22) (2016) 13413–13453.
[27] B. Zhang, H. Wang, S. Shen, X. She, W. Shi, J. Chen, Q. Zhang, Y. Hu, Z. Pang, X.
Jiang, Fibrin-targeting peptide CREKA-conjugated multi-walled carbon
nanotubes for self-amplified photothermal therapy of tumor, Biomaterials
79 (2016) 46–55.
[28] M. Tarfaoui, K. Lafdi, A. El Moumen, Mechanical properties of carbon
nanotubes based polymer composites, Compos. Part B-Eng. 103 (2016) 113–
121.
[29] S.R. Shin, S.M. Jung, M. Zalabany, K. Kim, P. Zorlutuna, S.B. Kim, M. Nikkhah, M.
Khabiry, M. Azize, J. Kong, Carbon-nanotube-embedded hydrogel sheets for
engineering cardiac constructs and bioactuators, ACS Nano 7 (3) (2013) 2369–
2380.
[30] M. Kharaziha, S.R. Shin, M. Nikkhah, S.N. Topkaya, N. Masoumi, N. Annabi, M.R.
Dokmeci, A. Khademhosseini, Tough and flexible CNT–polymeric hybrid
scaffolds for engineering cardiac constructs, Biomaterials 35 (26) (2014)
7346–7354.
[31] Q. Wan, M. Liu, J. Tian, F. Deng, G. Zeng, Z. Li, K. Wang, Q. Zhang, X. Zhang, Y.
Wei, Surface modification of carbon nanotubes by combination of mussel
inspired chemistry and SET-LRP, Polym. Chem. 6 (10) (2015) 1786–1792.
[32] N. Saito, H. Haniu, Y. Usui, K. Aoki, K. Hara, S. Takanashi, M. Shimizu, N. Narita,
M. Okamoto, S. Kobayashi, Safe clinical use of carbon nanotubes as innovative
biomaterials, Chem. Rev. 114 (11) (2014) 6040–6079.
[33] N. Karousis, N. Tagmatarchis, D. Tasis, Current progress on the chemical
modification of carbon nanotubes, Chem. Rev. 110 (9) (2010) 5366–5397.
[34] A. Eitan, K. Jiang, D. Dukes, R. Andrews, L.S. Schadler, Surface modification of
multiwalled carbon nanotubes: toward the tailoring of the interface in
polymer composites, Chem. Mater. 15 (16) (2003) 3198–3201.
[35] H. Lee, S.M. Dellatore, W.M. Miller, P.B. Messersmith, Mussel-inspired surface
chemistry for multifunctional coatings, Science 318 (5849) (2007) 426–430.
[36] N.W.S. Kam, M. O’Connell, J.A. Wisdom, H. Dai, Carbon nanotubes as
multifunctional biological transporters and near-infrared agents for selective
cancer cell destruction, PNAS 102 (33) (2005) 11600–11605.
[37] Q. Xin, H. Shah, A. Nawaz, W. Xie, M.Z. Akram, A. Batool, L. Tian, S.U. Jan, R.
Boddula, B. Guo, Q. Liu, J.R. Gong, Antibacterial carbon-based nanomaterials,
Adv. Mater. (2018) 1804838.
[38] O. Meincke, D. Kaempfer, H. Weickmann, C. Friedrich, M. Vathauer, H. Warth,
Mechanical properties and electrical conductivity of carbon-nanotube filled
polyamide-6 and its blends with acrylonitrile/butadiene/styrene, Polymer 45
(3) (2004) 739–748.
[39] J.L. Wang, K.F. Ren, H. Chang, S.M. Zhang, L.J. Jin, J. Ji, Facile fabrication of robust
superhydrophobic multilayered film based on bioinspired poly(dopamine)-
modified carbon nanotubes, Phys. Chem. Chem. Phys. 16 (7) (2014) 2936–
2943.
[40] Z. Deng, Y. Guo, X. Zhao, P.X. Ma, B. Guo, Multifunctional stimuli-responsive
hydrogels with self-healing, high conductivity, and rapid recovery through
host–guest interactions, Chem. Mater. 30 (5) (2018) 1729–1742.
[41] Z. Deng, T. Hu, Q. Lei, J. He, P.X. Ma, B. Guo, Stimuli-responsive conductive
nanocomposite hydrogels with high stretchability, self-healing, adhesiveness,
and 3D printability for human motion sensing, ACS Appl. Mater. Interfaces 11
(7) (2019) 6796–6808.
[42] X. Zhao, R. Dong, B. Guo, P.X. Ma, Dopamine-incorporated dual bioactive
electroactive shape memory polyurethane elastomers with physiological
shape recovery temperature, high stretchability, and enhanced C2C12
myogenic differentiation, ACS Appl. Mater. Interfaces 9 (35) (2017) 29595–
29611.
[43] X. Zhao, H. Wu, B. Guo, R. Dong, Y. Qiu, P.X. Ma, Antibacterial anti-oxidant
electroactive injectable hydrogel as self-healing wound dressing with
hemostasis and adhesiveness for cutaneous wound healing, Biomaterials 122
(2017) 34–47.
[44] J. Qu, X. Zhao, P.X. Ma, B. Guo, pH-responsive self-healing injectable hydrogel
based on N-carboxyethyl chitosan for hepatocellular carcinoma therapy, Acta
Biomater. 58 (2017) 168–180.
[45] B. Guo, P.X. Ma, Synthetic biodegradable functional polymers for tissue
engineering: a brief review, Sci. China Chem. 57 (4) (2014) 490–500.
[46] C. Shao, M. Wang, L. Meng, H. Chang, B. Wang, F. Xu, J. Yang, P. Wan, Mussel-
inspired cellulose nanocomposite tough hydrogels with synergistic self-
healing, adhesive, and strain-sensitive properties, Chem. Mater. 30 (9)
(2018) 3110–3121.
[47] R. Gharibi, H. Yeganeh, A. Rezapour-Lactoee, Z.M. Hassan, Stimulation of
wound healing by electroactive, antibacterial, and antioxidant polyurethane/
siloxane dressing membranes: In vitro and in vivo evaluations, ACS Appl.
Mater. Interfaces 7 (43) (2015) 24296–24311.
[48] R. Dong, X. Zhao, B. Guo, P.X. Ma, Self-healing conductive injectable hydrogels
with antibacterial activity as cell delivery carrier for cardiac cell therapy, ACS
Appl. Mater. Interfaces 8 (27) (2016) 17138–17150.
[49] Y. Liang, X. Zhao, P.X. Ma, B. Guo, Y. Du, X. Han, pH-responsive injectable
hydrogels with mucosal adhesiveness based on chitosan-grafted-
dihydrocaffeic acid and oxidized pullulan for localized drug delivery, J.
Colloid Interf. sci. 536 (2019) 224–234.
[50] J. Qu, X. Zhao, Y. Liang, T. Zhang, P.X. Ma, B. Guo, Antibacterial adhesive
injectable hydrogels with rapid self-healing, extensibility and compressibility
527
as wound dressing for joints skin wound healing, Biomaterials 183 (2018)
185–199.
[51] J.J. Bae, J.H. Yoon, S. Jeong, B.H. Moon, J.T. Han, H.J. Jeong, G.-W. Lee, H.R.
Hwang, Y.H. Lee, S.Y. Jeong, Sensitive photo-thermal response of graphene
oxide for mid-infrared detection, Nanoscale 7 (38) (2015) 15695–15700.
[52] X. Zhao, B. Guo, H. Wu, Y. Liang, P.X. Ma, Injectable antibacterial conductive
nanocomposite cryogels with rapid shape recovery for noncompressible
hemorrhage and wound healing, Nat. Commun. 9 (2018) 2784.
[53] H. Xu, Z. Fang, W. Tian, Y. Wang, Q. Ye, L. Zhang, J. Cai, Green fabrication of
amphiphilic quaternized b-chitin derivatives with excellent biocompatibility
and antibacterial activities for wound healing, Adv. Mater. 30 (29) (2018)
1801100.
[54] M.-C. Wu, A.R. Deokar, J.-H. Liao, P.-Y. Shih, Y.-C. Ling, Graphene-based
photothermal agent for rapid and effective killing of bacteria, ACS Nano 7 (2)
(2013) 1281–1290.
[55] C.J. Tormos, C. Abraham, S.V. Madihally, Improving the stability of chitosan–
gelatin-based hydrogels for cell delivery using transglutaminase and
controlled release of doxycycline, Drug Deliv. Transl. Re. 5 (6) (2015) 575–584.
[56] B.L. Guo, J.F. Yuan, Q.Y. Gao, pH and ionic sensitive chitosan/carboxymethyl
chitosan IPN complex films for the controlled release of coenzyme A, Colloid
Polym. Sci. 286 (2) (2008) 175–181.
[57] J. Qu, X. Zhao, P.X. Ma, B. Guo, Injectable antibacterial conductive hydrogels
with dual response to an electric field and pH for localized ‘‘smart” drug
release, Acta Biomater. 72 (2018) 55–69.
[58] S.J. Ryu, H. Jung, J.M. Oh, J.K. Lee, J.H. Choy, Layered double hydroxide as novel
antibacterial drug delivery system, J. Phys. Chem. Solids 71 (4) (2010) 685–
688.
[59] C. Wiegand, M. Abel, P. Ruth, P. Elsner, U.-C. Hipler, In vitro assessment of the
antimicrobial activity of wound dressings: influence of the test method
selected and impact of the pH, J. Mater. Sci.: Mater. Med. 26 (1) (2015) 18.
[60] A. Sasidharan, L.S. Panchakarla, A.R. Sadanandan, A. Ashokan, P. Chandran, C.M.
Girish, D. Menon, S.V. Nair, C. Rao, M. Koyakutty, Hemocompatibility and
macrophage response of pristine and functionalized graphene, Small 8 (8)
(2012) 1251–1263.
[61] X. Zhao, P. Li, B. Guo, P.X. Ma, Antibacterial and conductive injectable
hydrogels based on quaternized chitosan-graft-polyaniline/oxidized dextran
for tissue engineering, Acta Biomater. 26 (2015) 236–248.
[62] Z. Fan, B. Liu, J. Wang, S. Zhang, Q. Lin, P. Gong, L. Ma, S. Yang, A novel wound
dressing based on Ag/graphene polymer hydrogel: effectively kill bacteria and
accelerate wound healing, Adv. Funct. Mater. 24 (25) (2014) 3933–3943.
[63] J. Qu, X. Zhao, Y. Liang, Y. Xu, P.X. Ma, B. Guo, Degradable conductive injectable
hydrogels as novel antibacterial, anti-oxidant wound dressings for wound
healing, Chem. Eng. J. 362 (2019) 548–560.
[64] Y. Liang, X. Zhao, T. Hu, B. Chen, Z. Yin, P.X. Ma, B. Guo, Adhesive hemostatic
conducting injectable composite hydrogels with sustained drug release and
photothermal antibacterial activity to promote full-thickness skin
regeneration during wound healing, Small 15 (2019) 1900046.
[65] X. Zhao, M. Zhang, B. Guo, P.X. Ma, Mussel-inspired injectable supramolecular
and covalent bond crosslinked hydrogels with rapid self-healing and recovery
properties via a facile approach under metal-free conditions, J. Mater. Chem. B
4 (41) (2016) 6644–6651.
[66] B. Guo, J. Qu, X. Zhao, M. Zhang, Degradable conductive self-healing hydrogels
based on dextran-graft-tetraaniline and N-carboxyethyl chitosan as injectable
carriers for myoblast cell therapy and muscle regeneration, Acta Biomater. 84
(2019) 180–193.
[67] L. Wang, Y. Wu, T. Hu, B. Guo, P.X. Ma, Electrospun conductive nanofibrous
scaffolds for engineering cardiac tissue and 3D bioactuators, Acta Biomater. 59
(2017) 68–81.
[68] D. Gopinath, M.R. Ahmed, K. Gomathi, K. Chitra, P.K. Sehgal, R. Jayakumar,
Dermal wound healing processes with curcumin incorporated collagen films,
Biomaterials 25 (10) (2004) 1911–1917.
[69] M. Li, J. Chen, M. Shi, H. Zhang, P.X. Ma, B. Guo, Electroactive anti-oxidant
polyurethane elastomers with shape memory property as non-adherent
wound dressing to enhance wound healing, Chem. Eng. J. 375 (2019)
121999.
[70] D.-Y. Zhang, Y. Zheng, H. Zhang, J.-H. Sun, C.-P. Tan, L. He, W. Zhang, L.-N. Ji, Z.-
W. Mao, Delivery of phosphorescent anticancer iridium(III) complexes by
polydopamine nanoparticles for targeted combined photothermal-
chemotherapy and thermal/photoacoustic/lifetime imaging, Adv. Sci. 5 (10)
(2018) 1800581.
[71] X. Wang, C. Wang, X. Wang, Y. Wang, Q. Zhang, Y. Cheng, A polydopamine
nanoparticle-knotted poly(ethylene glycol) hydrogel for on-demand drug
delivery and chemo-photothermal therapy, Chem. Mater. 29 (3) (2017) 1370–
1376.
[72] A.C.D.O. Gonzalez, T.F. Costa, Z.D.A. Andrade, A.R.A.P. Medrado, Wound healing
- a literature review, An. Bras. Dermatol. 91 (2016) 614–620.
[73] C.L. Baum, C.J. Arpey, Normal cutaneous wound healing: clinical correlation
with cellular and molecular events, Dermatol. Surg. 31 (6) (2005) 674–686.
[74] T.K. Hunt, Wound Healing and Wound Infection: Theory and Surgical Practice,
Appleton-Century-Crofts, New York, 1980.
[75] M.K. Lichtman, M. Otero-Vinas, V. Falanga, Transforming growth factor beta
(TGF-b) isoforms in wound healing and fibrosis, Wound Rep. Reg. 24 (2) (2016)
215–222.
[76] S. Sanjabi, L.A. Zenewicz, M. Kamanaka, R.A. Flavell, Anti-inflammatory and
pro-inflammatory roles of TGF-b, IL-10, and IL-22 in immunity and
autoimmunity, Curr. Opin. Pharmacol. 9 (4) (2009) 447–453.
528 Y. Liang et al. / Journal of Colloid and Interface Science 556 (2019) 514–528
[77] J.W. Penn, A.O. Grobbelaar, K.J. Rolfe, The role of the TGF-b family in wound
healing, burns and scarring: a review, Int. J. Burn. Trauma 2 (1) (2012) 18–28.
[78] M. Chen, J. Tian, Y. Liu, H. Cao, R. Li, J. Wang, J. Wu, Q. Zhang, Dynamic covalent
constructed self-healing hydrogel for sequential delivery of antibacterial agent
and growth factor in wound healing, Chem. Eng. J. 373 (2019) 413–424.
[79] M. Shi, H. Zhang, T. Song, X. Liu, Y. Gao, J. Zhou, Y. Li, Sustainable dual release of
antibiotic and growth factor from pH-responsive uniform alginate composite
microparticles to enhance wound healing, ACS Appl. Mater. Interfaces 11 (25)
(2019) 22730–22744.
[80] F. Zehtabi, P. Ispas-Szabo, D. Djerir, L. Sivakumaran, B. Annabi, G. Soulez, M.A.
Mateescu, S. Lerouge, Chitosan-doxycycline hydrogel: an MMP inhibitor/
sclerosing embolizing agent as a new approach to endoleak prevention and
treatment after endovascular aneurysm repair, Acta Biomater. 64 (2017) 94–
105.
[81] J. Zhou, D. Yao, Z. Qian, S. Hou, L. Li, A.T.A. Jenkins, Y. Fan, Bacteria-responsive
intelligent wound dressing: simultaneous In situ detection and inhibition of
bacterial infection for accelerated wound healing, Biomaterials 161 (2018) 11–
23.
[82] Q. Shi, X. Luo, Z. Huang, A.C. Midgley, B. Wang, R. Liu, D. Zhi, T. Wei, X. Zhou, M.
Qiao, J. Zhang, D. Kong, K. Wang, Cobalt-mediated multi-functional dressings
promote bacteria-infected wound healing, Acta Biomater. 86 (2019) 465–479.