Feature Article
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Transparent Conductive Supramolecular Hydrogels with
Stimuli-Responsive Properties for On-Demand Dissolvable
Diabetic Foot Wound Dressings
Yue Zhao, Zuhao Li, Qiuju Li, Longfei Yang, Hou Liu, Ruyue Yan, Lizhi Xiao, He Liu,*
Jingcheng Wang,* Bai Yang, and Quan Lin*
Diabetic foot ulcers (DFU) remain a very considerable health care burden,
and their treatment is difficult. Hydrogel-based wound dressings are
appealing to provide an optimal environment for wound repair. However,
the currently available hydrogel dressings still need surgical or mechanical
debridement from the wound, causing reinjury of the newly formed tissues,
wound infection, delayed healing time, and personal suffering. Addition-
ally, to meet people’s increasing demand, hydrogel wound dressings with
improved performance and multifunctionality are urgently required. Here,
a new multifunctional supramolecular hydrogel for on-demand dissolvable
diabetic foot wound dressings is designed and constructed. Based on multi-
hydrogen bonds between hydrophilic polymers, the resultant supramolecular
hydrogels present controlled and excellent properties, such as good transpar-
ency, antibacterial ability, conductive, and self-healing properties. Thus, the
supramolecular hydrogels improve the new tissue formation and provide a
significant therapeutic effect on DFU by inducing angiogenesis, enhancing
collagen deposition, preventing bacterial infection, and controlling wound
infection. Remarkably, the resultant hydrogels also exhibit stimuli-responsive
ability, which renders its capability to be dissolved on-demand, allowing for a
facile DFU dressing removal. This multifunctional supramolecular hydrogel
may provide a novel concept in the design of on-demand dissolvable wound
dressings.
Dr. Y. Zhao, Dr. H. Liu, Prof. B. Yang, Prof. Q. Lin
State Key Lab of Supramolecular Structure and Materials, College of
Chemistry
Jilin University
Changchun 130012, P. R. China
E-mail: linquan@jlu.edu.cn
Dr. Z. Li, Q. Li, Dr. L. Yang, Dr. H. Liu, Prof. J. Wang
Orthopaedic Medical Center
The Second Hospital of Jilin University
Changchun 130041, P. R. China
E-mail: heliu@jlu.edu.cn; wangjinc@jlu.edu.cn
R. Yan, L. Xiao
Jilin Ginseng Academy
Changchun University of Chinese Medicine
Changchun 130117, P. R. China
The ORCID identification number(s) for the author(s) of this article
can be found under https://doi.org/10.1002/marc.202000441.
DOI: 10.1002/marc.202000441
Macromol. Rapid Commun. 2020, 2000441 2000441  (1 of 11)
1. Introduction
Diabetic foot ulcer (DFU) is a common
complication in patients with diabetic
hyperglycemia, which leaves patients at
increased high risk of morbidity, infec-
tion, nontraumatic limb amputations,
and even early death.[1–5] Among various
efforts to address this urgent issue, wound
dressings are effective strategies to pro-
vide an optimal environment for wound
repair.[6,7] Traditional wound dressings
for DFU treatment typically cover rubber,
electrospun nanofiber, cotton wool, nat-
ural or synthetic bandages, and gauzes.[8,9]
Although these dry dressings are conven-
ient to control the initial state of wound
healing, they tend to adhere to the wound
area once the absorbed blood and exudate
dry out.[7] Besides, the dry dressings still
suffer from limitations of maintaining a
moisture environment, allowing gaseous
exchange, and preventing infection.[6,7] In
this context, a wound dressing that can
deal with the above shortcomings would
be ideal to speed up diabetic foot wounds
healing and improve treatment outcomes.
In recent years, hydrogels have gener-
ated tremendous interest in wound healing applications.[10–16]
As water-based soft materials, hydrogels can facilitate wound
healing by absorbing wound exudate, preventing wound desic-
cation, and isolating the wound from the environment, which
makes them the best choice for wound healing.[17,18] However,
the currently available hydrogel dressings still require to be
changed frequently, which is a laborious process and inevitably
cause reinjury of the wounds, wound infection, delayed healing
time, and personal suffering.[19] To this end, on-demand dis-
solvable hydrogels represent a new class of emerging “smart”
wound dressings that can be readily operated and painlessly
removed.[19–24] Generally, this type of hydrogels can form in
situ and dissolve on-demand via physical crosslinking cases
and chemical crosslinking cases. The dissolution of physi-
cally crosslinked hydrogels is based on physical interactions,
such as molecular entanglements and/or secondary forces
(e.g., ionic, H-bonding, and hydrophobic associations).[19,20,22,23]
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These hydrogels possess dynamic networks that can be formed
and destroyed in response to environmental stimuli such as a
change in pH, magnetic, temperature, ionic strength of the
solution, or light. The dissolution of chemical crosslinked
hydrogels is achieved by using cleavable moieties that undergo
ester hydrolysis or enzymatic degradation. There has also
been growing interest in the preparation of dissolvable chemi-
cally crosslinked hydrogels by incorporation of thiol–disulfide
exchange, retro-Michael-type, and retro-Diels-Alder reactions,
and so on.[21] Despite much progress on on-demand dissolvable
hydrogels have been achieved, the majority of them are involved
with the complicated preparation procedures or the introduction
of potentially toxic crosslinking agents, which may greatly thwart
the large-scale fabrication of on-demand dissolvable hydrogel
dressings. Additionally, the function of developed on-demand
dissolvable hydrogel dressings is too simple that they could meet
people’s increasing demand for wound dressings with improved
performance and multifunctionality. When the on-demand dis-
solvable hydrogel dressings are applied for practical application,
it is desirable to maintain their basic properties while having
other enhanced characteristics, such as transparency, antibacte-
rial ability, conductivity, self-healing properties, and so on.
In this study, we developed a new transparent conductive
supramolecular hydrogel based on a multihydrogen bond net-
work structure, abbreviated as PVA-CEC-AGA/Ag (polyvinyl
alcohol-N-carboxyethyl chitosan-agarose/Ag). The resultant
supramolecular hydrogel not only possesses good transpar-
ency, antibacterial ability, conductive, and self-healing proper-
ties, but also exhibit stimuli-responsive ability. The macroscopic
structure of stimuli-responsive hydrogels can undergo gel-sol
transformation triggered by physical stimulus (temperature or
ultrasound) or chemical stimulus (pH or diol-containing mole-
cules). In addition, the PVA-CEC-AGA/Ag hydrogels are non-
toxic and have good in vitro cytocompatibility. These properties
render the supramolecular hydrogel useful for on-demand
Scheme 1.  The synthesis procedures of transparent conductive supramolecular hydrogel and its multinetwork structures.
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dissolvable wound dressings, allowing for a facile DFU wound
dressing removal. In vivo results suggest that this hydrogel
can effectively induce angiogenesis, enhance collagen deposi-
tion, prevent bacterial infection, and control wound infection in
DFU animal models, which in turn promote wound recovery.
Therefore, the transparent conductive supramolecular hydro-
gels can act as an on-demand dissolvable wound dressing for
wound repair, which may also have a positive therapeutic effect
on other types of wounds beyond DFU.
2. Results and Discussion
2.1. Fabrication and Characterization of the
PVA-CEC-AGA/Ag Hydrogel
The synthesis procedures of transparent conductive supramo-
lecular hydrogel and its multinetwork structures are depicted
in Scheme 1. To allow proper wound healing, we firstly selected
hydrogel components that are safe and compatible with the
biological system, and hydrogels with multinetwork structures
were directly fabricated. In brief, the solution of Na2B4O7 was
injected into the mixture solution of PVA, agarose, CEC, and
silver nanowires (Ag NWs) (Figure S1, Supporting Information)
under vigorously stirring at 90 °C. The transparent conductive
supramolecular hydrogel was instantly formed. The backbone
networks of hydrogels were attributed to the strong hydrogen
bonds (hydrogen bond I) between N-carboxyethyl chitosan-aga-
rose, N-carboxyethyl chitosan-PVA, and agarose-PVA.[25] These
strong hydrogen bonds endow hydrogels with stable mechanical
properties. In addition, the reversible networks were also gener-
ated due to the dynamic reactions of PVA-borax-PVA and PVA-
glycerol-PVA (hydrogen bonds II).[26] It is worth noting that the
conductive Ag NWs were uniformly dispersed in the hydrogel
because of the static hydrogen bond network (hydrogen
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bond III) between PVA and the nanowires. The sol-to-gel phase
transformation of hydrogels is displayed in Figure  1a, named
PVA-CEC-AGA/Ag. One of the essential requirements for
hydrogel wound dressings is their porosity.[27,28] The porosity
of wound dressings promises the transport of nutrition and
gases, thereby enhancing cell migration and proliferation. The
porosity and microarchitectural features of hydrogels were
observed by scanning electron microscope (SEM). Overall,
based on multihydrogen bonds between hydrophilic polymers,
the supramolecular hydrogels showed a heterogeneous struc-
ture (Figure  1b) with pore diameters ranging from 2.5 to 45
µm (Figure S2, Supporting Information). These porosities on
hydrogels could enable cellular penetration, which are signifi-
cant to the new tissue formation.
The swelling ability was measured by immersing the PVA-
CEC-AGA/Ag hydrogels into an excess of phosphate buffered
saline (PBS) (pH  =  7.4) for different periods of time.[19,29,30]
As illustrated in Figure  1c, the hydrogel swelled to 200% in
180  min and 250% in 360  min. After reaching equilibrium,
the hydrogel was stable and retained its integrity. These results
indicated that the swelling could not cause the breakage or dis-
solution of hydrogels. In addition, the hydrogel could contain
a large amount of water, which is beneficial to remove wound
exudate and maintain a moist environment for the wound beds.
2.2. Transparency of the PVA-CEC-AGA/Ag Hydrogel
Transparency is also an important requirement for hydrogel
dressings when doctors want to monitor the condition of
wounds and place dressings on the wound area for a long
time.[31–34] Figure 2a exhibits the plots of the transmittance of
light in the wavelength range 400–700  nm for hydrogels with
different concentrations of Ag NWs. The hydrogel’s transpar-
ency could be altered by changing the concentration of Ag
NWs. The average transmittance per millimeter of thickness of
the hydrogel at 660 nm wavelength decreased from 93% to 74%,
when the Ag NWs content increased from 0 to 0.2 mg mL−1. In
addition, when hydrogels were covered on a cardboard, the text
of “SKLJLU” on the cardboard was clearly observed indicating
their high transparency (Figure  2b). This may be due to Ag
NWs are well-dispersed in the hydrogel to form light-passible
nanoscale networks, which allows light to pass through and
endows hydrogel with high transparency.
2.3. Mechanical Strength and Self-Healing Performance of the
PVA-CEC-AGA/Ag Hydrogel
The mechanical properties of PVA-CEC-AGA/Ag could be easily
adjusted by varying the individual components (Figure S3,
Supporting Information). We constructed the resultant supra-
molecular hydrogels with crosslinking networks based on mul-
tihydrogen bonds to provide the tunable mechanical properties.
In addition, the hydrogen bonds between PVA-borax-PVA and
PVA-glycerol-PVA can be reversibly associated and disassoci-
ated at room temperature, thereby providing good self-healing
properties.[26] To evaluate the self-healing property of hydrogels,
a strain amplitude sweep test was first performed. As shown in
Figure  3a, in the low strain region, both the storage modulus
(G′) and loss modulus (G″) maintained constant values. With
enhanced strain, G′ and G″ curves intersected at a strain of
13.5%, which is the critical strain value indicating the transition
of the gel network to a liquid state. To further increase the

Figure 1.  a) Digital photos of the gelation progress. The mixture of prepared Ag NWs, PVA, and chitosan solution presented in the sol state, and it
experienced sol-gel transition after adding Na2B4O7. b) SEM images of the hydrogel. c) The swelling kinetics of the hydrogel incubated in distilled
water for different times. The hydrogel was prepared with PVA (5 wt%), agarose (0.025 g), CEC (0.025 g), glycerin (0.2 mL), Ag NWs (0.2 mg mL−1),
and Na2B4O7 (0.04 mol L−1, 10 mL).

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Figure 2.  a) Light transmittance spectra of the hydrogel with different concentration of Ag NWs in air. b) Optical images of the hydrogel with different
concentration of Ag NWs. The hydrogel was prepared with PVA (5 wt%), agarose (0.025 g), CEC (0.025 g), glycerin (0.2 mL), Ag NWs (0, 0.05, 0.10,
0.15, 0.20 mg mL−1), and Na2B4O7 (0.04 mol L−1, 10 mL).

strain to 100%, both G′ and G″ decreased dramatically because the self-healing behaviors of hydrogels. The continuous shear
of the collapse of the hydrogel networks. strain was alternated between 200% and 0.01% for 150 s at a
Based on the results of oscillatory shear rheology, the step- fixed angular frequency of 10 rad s−1, respectively. As displayed
strain sweeps were then performed to quantitatively measure in Figure  3b, when the hydrogel was subjected at a higher

Figure 3.  a) Strain-dependent oscillatory shear rheology of the PVA/chitosan/Ag hydrogel hydrogels at a fixed angular frequency (10 rad s−1). b) Using
alternating low (1%)–high (500%) shear strain to measure the self-healing property of PVA/chitosan/Ag hydrogel. Each strain interval was kept as
150 s and the fixed frequency was kept as 10 rad s−1. c) The hydrogels were cut into two pieces and then healed under ambient conditions. Rhodamine
B and methylene blue were present in the hydrogel for visualization. d) Macroscopic photographs of the restored electrical conductivity by a healed
hydrogel connected to purple LED bulbs. Methylene orange and blue were present in the hydrogel for visualization. The hydrogel was prepared with
PVA (10 wt%), agarose (0.025 g), CEC (0.025 g), glycerin (0.2 mL), Ag NWs (0.20 mg mL−1), and Na2B4O7 (0.04 mol L−1, 10 mL).

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strain of 200%, the gel failure occurred where G″ was larger
than G′. Subsequently, when the hydrogel was applied with a
lower strain of 0.01%, the G′ and G″ values were instantane-
ously fully restored to their original values even after repeated
multiple times. The mechanical healing ability of hydrogels was
also demonstrated by the macroscopic observation. According
to Figure  3c, the hydrogels with different colors were physi-
cally cut into two halves of the same size. Then, the different
parts of the hydrogel were put together. After 5  min, the two
halves of the hydrogels could be automatically reconstructed,
and they could maintain integrity. Apart from the mechanical
self-healing property, the electrical properties of the hydrogel
could also be restored. According to Figure 3d, when the hydro-
gels were cut into two parts, the LED bulbs were extinguished.
Afterward, the separated two parts were brought into contact
at room temperature. After 2 h, we found that the hydrogels
healed completed with an indistinguishable interface, and the
LED bulbs could be lighted up again.
2.4. Stimuli-Responsive and On-Demand Dissolvable Abilities of
the PVA-CEC-AGA/Ag Hydrogel
Intelligent soft materials have attracted increasing attention
during the past decades whose macroscopic structure and
physical properties show a significant change in response to
specific stimuli.[35–39] With the reversible linkages in hydrogels
demonstrated above, the prepared hydrogels are promising as
stimuli-responsive macroscopic structures. As illustrated in
Figure 4a, the hydrogel network could be completely destroyed

Figure 4.  a) The gel-sol phase transitions of the hydrogel triggered by different stimuli, such as fructose, glucose, dopamine, and Vitamin B6 and
HCl (0.1 m). b) Schematic of hydrogel’s multiresponses to different stimuli. c) Photographs of the on-demand dissolution of hydrogel after treatment
with a glucose solution (0.1 m, pH = 7.0). The hydrogel was prepared with PVA (10 wt%), agarose (0.025 g), CEC (0.025 g), glycerin (0.2 mL), Ag NWs
(0.2 mg mL−1), and Na2B4O7 (0.04 mol L−1, 10 mL).

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Figure 5.  a) Proliferation of L929 cells cultured for 1, 4, and 7 d (n = 3). b) Live/Dead staining of L929 cells for 3 d. The hydrogel was prepared with PVA
(5, 7.5, 10, 12.5 wt%), agarose (0.025 g), CEC (0.025 g), glycerin (0.2 mL), Ag NWs (0.2 mg mL−1), and Na2B4O7 (0.04 mol L−1, 10 mL).
by increasing the temperature or applying ultrasound. In addi-
tion to physical stimulation, reducing solution pH or adding
competitive diol-containing molecules (such as fructose, glu-
cose, dopamine, and vitamin B6) could also quickly trigger the
breakdown of the hydrogel. This stimuli-responsive property
was mainly due to the reversible nature of boronate formation
between Na2B4O7 and PVA. More specifically, the equilibrium
of boronate-based reactions could be shifted by heating, ultra-
sound, reducing pH, and diol-containing molecules. The diol
groups in free molecules are more reactive than PVA polymer,
which can act as a competitor of PVA to react with the Na2B4O7
(Figure  4b).[40] Therefore, the PVA-CEC-AGA/Ag hydrogels
break down and dissolve in the presence of molecules con-
taining diol groups (such as fructose, glucose, dopamine, and
vitamin B6).
Usually, traditional available dressings will adhere to the
wound surface, requiring surgical and mechanical removal
from the wound. The repeated dressing changes not only cause
additional trauma to the newly formed tissues but also bring
personal suffering in the injured patient.[21–23] Thus, hydrogels
with on-demand dissolvable property are particularly appealing
to improve treatments and reduce patient pain. It is anticipated
the PVA-CEC-AGA/Ag hydrogels have the capability to be dis-
solved on-demand via the reversible nature of boronate forma-
tion. The on-demand dissolution of the developed hydrogels
by glucose was investigated. As displayed in Figure  4ci,ii, the
PVA-CEC-AGA/Ag hydrogels were coated in an artificial wound
on a diabetic rat. After a glucose-soaked gauze was applied to
half of the hydrogel for about 20 min, the covered hydrogel
part dissolution occurred (Figure 4ciii,iv). If the glucose-soaked
gauze was administered to the whole hydrogel, the hydrogel
completely dissolved as time elapsed (Figure  4cv–viii). The
on-demand dissolvable capability of the hydrogels was further
quantitatively analyzed by recording G′ and G″ in time sweep
mode (Figure S4, Supporting Information). Obviously, the
hydrogel remained stable when in contact with air. In contrast,
when in contact with glucose (0.1 m, pH  =  7.0), the G′ and G″
of the hydrogel decreased and G″ became larger than G′ indi-
cating that the hydrogel experienced a sol-gel transformation.
Thus, these results demonstrated the ability of the PVA-CEC-
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AGA/Ag hydrogel to be dissolved on-demand, which is prom-
ising as a desirable alternative to debridement of the dressing.
2.5. Biocompatibility Evaluation of the PVA-CEC-AGA/Ag
Hydrogel
The in vitro cytotoxicity test (Cell Counting Kit 8 (CCK-8) assay)
was carried out by using the fibroblast (L929) cell line, and the
corresponding results are illustrated in Figure  5a. The control
cells were incubated in a culture dish, and other hydrogel group
cells were incubated with different components of hydrogel
extract solutions. The L929 cells were proliferating with the pro-
longation of culture time, and there was no significant differ-
ence between each hydrogel group and the control group. The
CCK-8 assay results clearly demonstrated that the PVA-CEC-
AGA/Ag hydrogel had good biocompatibility. In addition, Live/
Dead staining revealed that L929 cells maintained excellent
viability whether in the hydrogel group or the control group
(Figure 5b). Thus, PVA-CEC-AGA/Ag hydrogels exhibited good
in vitro cytocompatibility, which have great potential for use in
biomedical applications.
2.6. PVA-CEC-AGA/Ag Hydrogel for Diabetic Foot Wound
Dressing
DFUs are always regarded as one of the most serious compli-
cations of diabetes associated with amputation. The impaired
DFUs typically always have features of extensive size, irregular
shape, difficult-to-access areas, massive wound exudate, etc.[7]
Based on this situation, the development of benign and efficient
diabetic foot wound dressings is urgent to be explored. Com-
pared with other traditional wound dressings, hydrogel-based
wound dressing has more ideal properties including removal
of wound exudate, providing a moist wound environment, pre-
vention of secondary infections, and promotion of cell adhesion
and proliferation.[11,13,16] In addition to the above advantages, our
designed PVA-CEC-AGA/Ag hydrogels can be easily detached
from the wound by an on-demand dissolvable property. This
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nonharmful change characteristic can create an optimal
environment for promoting diabetic foot wound healing. To
examine the PVA-CEC-AGA/Ag hydrogel treatment of infec-
tive diabetic foot wounds, we established diabetic rat models
by injecting streptozotocin (STZ). In order to observe the rate
of wound healing, the general wound closure results were
recorded and the residual wound area calculated. According
to Figure 6a, the wounds treated with hydrogel showed a rapid
contraction and healed in a well-organized manner, while there
was suppurative wound surface in the control group. In addi-
tion, the quantitative analysis showed that the healing rate of
the wounds treated with hydrogels was significantly faster than
that of the control group, especially at the late stage of treat-
ment (10, 15, and 20 d post-treatment) (p < 0.01) (Figure 6b). At
20 d post-treatment, the wounds covered with hydrogels were
almost fully healed, whereas about 35.73  ±  7.33% of unhealed
wound areas had remained in the control group. During the
whole experiment, the blood glucose level of rats was also

Figure 6.  a) Representative photos of the diabetic foots after treated with PBS and hydrogel. b) Quantification of wound residual area. c) Hematoxylin
and eosin (H&E) staining of wound tissues around foots. d) Quantification of inflammatory cells. e) Masson trichrome staining of wound tissues
around foots. f) Quantitative analysis of the percentage of collagen deposition. g) Neovascularization in granulation tissue was assessed with CD31
immunohistochemical staining. h) Quantification of blood vessels. i) Giemsa staining for bacteria in wound tissue (*P < 0.05, **P < 0.01, n = 3). The
hydrogel was prepared with PVA (10 wt%), agarose (0.025 g), CEC (0.025 g), glycerin (0.2 mL), Ag NWs (0.2 mg mL−1), and Na2B4O7 (0.04 mol L−1,
10 mL).

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measured, and there was a drop in the blood glucose level after
the wounds were treated with PVA-CEC-AGA/Ag hydrogel,
especially in the first 5 d after treatment (p < 0.05) (Figure S5,
Supporting Information).
To further distinguish the quality of regenerated skin in
the diabetic foot wounds, histological analysis was performed.
Consistent with the optical image results, H&E staining
(Figure  6c) revealed that wounds treated by PVA-CEC-AGA/
Ag hydrogel displayed well-organized granulation tissue for-
mation on the 5th day. On the 15th day, more granulation tis-
sues were observed in the hydrogel group. In contrast, necrotic
tissue, foreign bodies, less granulation tissues, and excessive
inflammatory cells were formed in the control group during
the whole healing process. Additionally, the aggregation and
activation of appropriate inflammatory cells are essential to the
transition from inflammation to repair phases, while excessive
inflammatory cells impair wound repair. In the control group,
a large number of inflammatory cells were observed at the
wound site on the 5th day, and the inflammation was still
serious on the 15th day. Compared with the control group, the
wounds covered with hydrogel showed a mild inflammatory
reaction (p < 0.05), which maintained an appropriate inflamma-
tory environment and did not cause a foreign body response
(Figure  6d). Collagen deposition is another critical factor for
wound contraction and scar formation.[41] Masson’s trichrome
(MT) staining was performed to detect the newborn collagen
deposition in the wound (Figure  6e,f). On the 5th day, almost
no collagen deposition was observed in the control group, while
limited collagen deposition occurred in the hydrogel group. On
the 15th day, the collagen deposition of the hydrogel group was
further increased and extended to the central of wounds with
regular arrangement, which was 1.75-fold higher than that of
the control group.
It is well-known that some crucial substances (e.g., growth
factors, oxygen, nutrients, etc.) are delivered and infiltrated into
the wounded tissue via blood vessels.[5] As a result, neovascu-
larization plays a critical role in the tissue remodeling process,
and wounds may fail to heal due to inadequate local vascular
supply. As shown in CD31 immunohistochemical staining,
more neovascularization (green arrows) was generated after
treated with hydrogel, which was 4.76-fold and 2.2-fold higher
than the control group on the 5th and 15th days post-treatment
(p < 0.05), respectively (Figure 6g,h).
Diabetic wounds are easily infected by bacteria and facili-
tated bacterial growth because of local high blood glucose
levels and exposure to air. An infectious environment exerts an
obvious negative impact on wound healing, even causing bac-
teremia.[41] Giemsa staining was used to investigate bacterial
contamination on the wound surface. As shown in Figure  6i,
the wounds in the control group had a large number of bac-
teria on the 5th and 15th days. However, in the hydrogel group,
owing to the antibacterial properties of Ag NWs, it was difficult
to find colonizing bacteria (Figure S6, Supporting Information).
Furthermore, to evaluate the spread of bacteria in the whole
body, the number of white blood cells (WBC) in blood, an indi-
cator of systemic inflammation, was also monitored (Figure
S7, Supporting Information). The WBC counts in the hydrogel
group were within the normal range (5.0–25.0  ×  109 L−1).
However, the counts in the control group were significantly
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increased on the 5th and 15th day, which were 31.47 ± 2.87 and
33.4  ±  3.85  ×  109  L−1, respectively. This result further demon-
strates that this effective sterilization dressing can avoid sys-
temic infection, and remarkably accelerate wound closure and
enhance the quality of the healed wound as described above.
3. Conclusions
In summary, we have designed a new transparent conductive
supramolecular hydrogel through multihydrogen bond net-
works using biocompatible macromonomers. The developed
hydrogels exhibit many desirable features including tunable
mechanical, good transparency, antibacterial ability, conduc-
tive, and self-healing properties. It is worth mentioning that
the resultant hydrogels can dissolve on-demand via the revers-
ible nature of boronate formation, which renders significant
promise for a more effective treatment for DFU patients. In
vitro results demonstrate that the resultant hydrogels are bio-
compatible. In vivo DFU animal experiments result further
demonstrates that this hydrogel dressing can induce angio-
genesis, enhance collagen deposition, avoid systemic infection,
remarkably accelerate wound closure, and enhance the quality
of the healed wound. This work provides a unique approach to
design and construct multifunctional hydrogels with controlled
dissolution ability.
4. Experimental Section
Materials: Silver nitrate (AgNO3, ≈99.8%) was purchased from
Sinopharm Chemical Reagent Co., Ltd. (Shanghai, P. R. China).
Hydrochloric acid (HCl, 36–38%), sodium hydroxide (NaOH, ≈99.0%),
ethanol (C2H5OH, ≈99.7%), ethylene glycol (≈99%), and sodium
tetraborate (Na2B4O7·10H2O) were purchased from Beijing Chemical
Works (Beijing, P. R. China). Polyvinyl pyrrolidone (PVPON, Mw ≈
40 000), PVA (≈99% hydrolyzed, Mw ≈ 130 000), dopamine hydrochloride
(DA, ≈98%), STZ, glucose, and fructose were obtained from
Sigma-Aldrich. Chitosan (degree of deacetylation ≈ 95%, 100–200 mps)
and vitamin B6 hydrochloride were purchased from Aladdin. Acrylic
acid (≈99%) and paraformaldehyde (≈99%) were from Xilong Chemical
Co., Ltd. (Guangzhou, P. R. China). Copper chloride (CuCl2·2H2O)
was from Tianjin Yongsheng Fine Chemical Co., Ltd. (Tianjin, P. R.
China). Dulbecco’s modified Eagle’s medium (DMEM, low glucose)
was purchased from HyClone (Beijing, P. R. China). Fetal bovine
serum (FBS) and streptomycin-penicillin were purchased from Gibco
Life Technologies (USA). CCK-8 was obtained from Dojindo Molecular
Technology (Japan). Calcein-AM/PI was obtained from Biobetimes
Biotechnology Co., Ltd. (Changsha, P. R. China). Hematoxylin and
eosin, Masson’s trichrome (MT), and Giemsa stain were purchased
from Thermo Fisher Scientific Co., Ltd. (Shanghai, P. R. China). Triton
X-100 was purchased from Dingguo Changsheng Biotechnology
Co. Ltd. (Beijing, P. R. China). Antibody CD31 (ab28364) used for
immunohistochemistry was purchased from Abcam (UK). The ultrapure
water used throughout the experiments was purified with a Milli-Q A10
filtration system (Millipore, Billerica, MA). The other chemicals were
used as purchased without further purification.
Characterization: Ultraviolet–visible (UV–vis) absorption spectra were
recorded at room temperature on a Lambad 800 spectrophotometer
using a quartz cell of 1  cm path length. Field-emission SEM images
were obtained with JEOL JSM-6700F SEM operated at an acceleration
voltage of 3  kV. The dynamic mechanical properties of hydrogels were
performed on a rheometer (TA Instruments-waters LLC) at room
temperature. The optical density of cells was measured at 450 nm by a
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www.advancedsciencenews.com
microplate reader (Multiskan EX, Thermo Fisher Scientific Inc., Shanghai,
P. R. China). Fluorescent photographs cell and histological staining
images were obtained from a fluorescence microscopy (IX51, Olympus,
Japan). The blood glucose level was measured by a glucose meter
(Johnson, P. R. China). All histological staining images were analyzed by
Image Pro-Plus (IPP) 6.0 (National Institutes of Health, Bethesda, MD).
All photographs were taken by a digital camera (Canon camera).
Preparation of Hydrogels: The hydrogels were synthesized by
the following procedures. 1) Preparation of Ag NWs: Ag NPs were
fabricated according to literature procedures.[42–45] Briefly, anhydrous
ethylene glycol (5 mL) was heated to 151.5 °C under magnetic stirring
(260  rpm). At 1 h, the solution of CuCl2 dissolved in ethylene glycol
(4  mmol L−1, 0.9  mL) was injected into the heated ethylene glycol.
After 15  min, the solution of PVPON dissolved in ethylene glycol
(0.1 g, 2.5 mL) was added, followed by injection of the ethylene glycol
solution of AgNO3 (0.05  g, 2.5  mL). The above solution was then
heated for another 30  min, and Ag NWs were synthesized. Products
were then washed with ethanol once and water three times. They were
stored in water before use. 2) Preparation of N-carboxyethyl chitosan:
CEC was fabricated following previous reported procedures. Chitosan
(2 g, 12.4 mmol) and acrylic acid (2.9 mL, 42.6 mmol) were dissolved
in deionized water (100 mL), and then heated at 50 °C under magnetic
stirring for 72 h. Afterward, the above solution was cooled down,
whose pH was adjusted to 10–12 by injecting 1 m NaOH. To remove
by-products and other impurities, the solution was dialyzed (MWCO
14  000) against deionized water for 3 d. After repeated change of
water, the product was freeze-dried. 3) Preparation of hydrogels: In
this experiment, PVA (0.5  g), agarose (0.025  g), CEC (0.025  g), and
glycerin (0.2  mL) were dissolved in deionized water (10  mL), and
this mixture was oil-bathed at 90  °C under magnetic stirring for 2  h.
Then different concentrations of Ag NWs (0, 0.05, 0.10, 0.15, and
0.2  mg  mL−1) were added into the above solution, respectively. After
stirred for another 15 min, Na2B4O7 (0.04 mol L−1, 10 mL) was injected
into the above mixture, and further stirred at 90 °C for 10 min until a
hybrid hydrogel formed.
Biocompatibility of Hydrogels In Vitro: The hydrogel samples were cut
into thin slices placed on six-well plates. The prepared disc samples
were added with DMEM containing 10% FBS, 1% penicillin and
streptomycin, and then incubated at 37  °C in a humidified atmosphere
containing CO2 (5%) for 3 d. The hydrogel extracts were subsequently
used for the following in vitro cell experiments. The proliferation of L929
cells on hydrogel extracts was measured by CCK-8. Briefly, L929 cells
at a density of 5 × 104 each well were cultured with hydrogel extracts.
For comparison, the control groups were cultured with DMEM. After
incubation for 1, 4, and 7 d, CCK-8 solution with a 10% volume of the
medium was added into each well after replacing medium and then were
incubated at 37 °C in a humidified atmosphere containing CO2 (5%) for
2 h. After that, the above reaction solution (100 µL) was transferred into
a new 96-well plate, and the optical density was measured at 450 nm by
a microplate reader.[10,13]
To evaluate the cell viability of L929 cultured in the hydrogel extracts,
the Live/Dead assay was performed. 3 d after seeding, samples were
incubated with 1  × 10−3 m calcein-AM for 1 h, then incubated with
1  μg  mL−1 propidium iodide (PI) for 5  min at 37  °C. The samples were
then imaged by fluorescence microscopy.[46]
Diabetic Rat Model Establishment and Wound Healing Examination:
Animal experiments were conducted according to the National Institutes
of Health’s Guide for the Care and Use of Laboratory Animals (NIH
Publications No. 8023, revised 1978). All experimental protocols were
approved and performed in accordance with the guidelines of the Animal
Ethics Committee of Jilin University. Adult male Sprague-Dawley (SD)
rats (200–250 g, 8–10 weeks) were obtained from the Animal Center of
Jilin University. The diabetes rats model was prepared by intraperitoneal
injection of STZ (55 mg kg−1). After one week of STZ administration, the
blood glucose level was measured by a glucose meter from the tail vein.
Rats were diabetic when their blood glucose level exceeded 16.7  mmol
L−1. Blood glucose levels of rats were monitored throughout the study.
Following by STZ-induced hyperglycemia for three weeks, 40 rats were
Macromol. Rapid Commun. 2020, 2000441 2000441  (9 of 11)
www.mrc-journal.de
selected to establish diabetic wound models. After anesthesia with
intraperitoneal 1% pentobarbital sodium (100  mg kg−1), rat skin was
cleaned with 0.3% iodine. Then a 5 mm diameter full-thickness foot skin
wound was made by punch biopsy instruments. The wound was washed
with saline, and implanted with Staphylococcus aureus and Escherichia
coli (1 × 108 CFUs in 10 × 10−6 m of PBS), respectively. For comparison,
the wound covered with 200 µL hydrogel was regarded as an experiment
group, while the wound just treated with PBS was regarded as a control
group. All the wounds were fixed with sterile gauze and fixed with an
elastic adhesive bandage. The rats were individually housed in a cage for
observation. At the 0, 5, 10, 15, and 20 d postoperation, the wounds were
recorded using a digital camera and quantitatively analyzed by Image
Pro Plus 6.0. The residual wound area was calculated by the following
formula: Residual wound area (%) = Sn/S0  × 100% (S0: initial wound
area; Sn: wound area at different time points).[41]
Histopathological Evaluation: After 5 and 15 d of treatment, the
wound and surrounding tissues were collected. All the skin samples
were fixed with 10% paraformaldehyde solution, and then embedded
in paraffin for routine histological processing. On the basis of the
standard protocols, 5  mm thickness sections of the samples were
prepared. In this experiment, H&E and MT staining were carried out to
assess the inflammation, morphology, tissue regeneration, and collagen
deposition. CD31 immunohistochemistry was applied to observe the
neovascularization in regenerated granulation tissue, while Giemsa
staining was used to investigate the bacterial colonization in wound
tissues.[19,47,48]
Statistical Analysis: The data were expressed as means ± standard
deviation (SD) from at least three independent experiments, and the
statistical analysis was carried out using ANOVA with Tukey’s posthoc
analysis (SPSS Inc., Chicago, IL). P < 0.05 was considered a statistically
significant difference.
Supporting Information
Supporting Information is available from the Wiley Online Library or
from the author.
Acknowledgements
Y.Z. and Z.L. contributed equally to this work. This work was supported
by the National Natural Science Foundation of China (Grant Nos.
51861145311, 21174048, 81671804, and 81772456). J.W. and H.L. are thankful
to the Science and Technology Development Program of Jilin Province
(Grant Nos. 20190304123YY, 20180623050TC, 20180201041SF, and
20170204004GX), the Cultivation Program from the Second Hospital of Jilin
University for National Natural Science Foundation (KYPY2018-01), and the
Youth Talents Promotion Project of Jilin Province (Grant No. 192004). The
authors thank the cooperation with Prof. Hongdong Li by Open Project of
State Key Laboratory of Super hard Materials (Jilin University).
Conflict of Interest
The authors declare no conflict of interest.
Keywords
dissolution, self-healing hydrogels, silver nanowires, stimuli-responsive
hydrogels, wound dressing
Received: August 12, 2020
Revised: September 3, 2020
Published online:
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www.advancedsciencenews.com
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Zuhao Li’s, main research focus on two main directions: hydrogels as wound dressing to enhance
diabetic wound healing, as well as 3D-printed metal scaffolds to promote bone integration in
various orthopedic related diseases.

He Liu is an attending physician at the Orthopedic Medical Center of The Second Hospital of Jilin
University. His main research direction is the application of hydrogel in different disease models.

Jincheng Wang is a professor, doctoral supervisor, and the Director of Orthopedic Medical
Center of The Second Hospital of Jilin University. His research interest is focused on the clinical
application of 3D printing technology.

Quan Lin received his bachelor’s degree in polymer materials and engineering and master’s
degree in polymer chemistry and physics from JLU in 1991 and 1994, respectively. He started his
professional experience at the College of Chemistry, JLU from 1994, and he graduated with a Ph.D.
in polymer chemistry and physics from JLU in 2000. He joined Carleton University as a postdoc-
toral researcher during 2001–2003. His research interests focus on multifunctional nanomaterials
based on supramolecular chemistry for applications in biosensing, responsive, and biomimetic
polymeric hydrogels and applications in biological systems.

Macromol. Rapid Commun. 2020, 2000441 2000441  (11 of 11) © 2020 Wiley-VCH GmbH