A wet electric material, its preparation method and application

Abstract

The application discloses a kind of wet electric materials and preparation method and application thereof, the preparation method of the wet electric material includes the following steps: adding PEDOT:PSS to the solution of gelatin and alginate and mixing, obtain MEG hydrogel solution;Ag nanoparticles and shiva are added to the solution of gelatin and alginate and mixed, and MEG hydrogel solution is obtained BFC hydrogel solution;With calcium chloride solution, MEG hydrogel solution and BFC hydrogel solution are carried out layered ion crosslinking gelation, then washing, freeze-drying, the wet electric material is obtained;The wet electric material prepared by the application does not need external power supply, the wet electric material spontaneously generates electric effect using humidity of wound environment, and the electroactive bacteria in the wet electric material generates hydrogen sulfide mediated gas therapy by metabolism, effectively controls wound inflammation, accelerates wound healing, solves the problem that bioelectronic device in the prior art relies on rigid power supply and lacks dynamic response capability.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, specifically to a wet electrochemical material, its preparation method, and its application. Background Technology

[0002] Human tissues generate weak "damage currents" or "damage potentials" at the site of skin injury. This endogenous electric field is a key signal guiding cells to migrate towards the wound and initiating the repair process. Based on this mechanism, active intervention using external electrical stimulation to simulate or enhance the body's own bioelectrical signals to regulate disordered repair processes and accelerate wound tissue regeneration has been widely reported. During wound healing, electrical stimulation plays a crucial role in promoting fibroblast migration, angiogenesis, and macrophage polarization. These events correspond to the formation of granulation tissue during the proliferative phase, the construction of new blood vessel networks, and the orderly transition from the inflammatory phase to the repair phase, respectively, and are essential for tissue regeneration. One of its mechanisms of action is that the electric field can affect the activity of cell signaling pathways. Calcium ions, as important intracellular second messengers, directly regulate cytoskeleton rearrangement, gene expression, and growth factor secretion through spatiotemporal changes in their concentration. Electrical stimulation regulates calcium ion (Ca) activity. 2+ The influx of calcium into the skin activates calcium signaling pathways, accelerating the epithelialization of new tissue and reducing scar formation. Clinically, electrical stimulation has been validated for its efficacy in promoting granulation tissue growth, reducing edema, controlling infection, and relieving pain. Furthermore, electrical stimulation combined with standard care, compared to conventional treatment alone, significantly shortens the healing time of chronic wounds and improves the complete healing rate. Wound management guidelines in various countries are gradually incorporating electrical stimulation into standard treatment recommendations as an important physical therapy for treating refractory wounds such as diabetic foot ulcers and pressure injuries. Currently, the application of electrical stimulation is evolving from purely in-hospital treatment to portable and intelligent solutions. Wearable micro-electrical stimulation patches and closed-loop feedback systems combined with smart dressings are becoming increasingly common, allowing patients to receive continuous treatment in the community or at home. This shift not only improves patient adherence but also provides a more accessible solution for the long-term management of chronic wounds.

[0003] Chronic, refractory wounds often fail to heal properly due to local ischemia, infection, or impaired cellular electrical signal transduction. Traditional methods such as debridement and dressings have limited effectiveness in addressing these complex pathological conditions. Therefore, significant clinical demand has prompted researchers to explore more effective physical therapy methods. Electrical stimulation, due to its non-invasive nature, fewer side effects, and ability to directly intervene in the core aspects of the repair mechanism, has gradually gained widespread attention. However, traditional bioelectronic platforms still face many limitations in practical applications. They often rely on rigid power sources, are bulky, and lack adaptability to soft, dynamic wound tissues; furthermore, most of these platforms lack the ability to dynamically respond to changes in pathological conditions, making it difficult to achieve precise control of the repair process. Summary of the Invention

[0004] The purpose of this invention is to provide a wet electro-material, its preparation method, and its application. The wet electro-material prepared by this invention does not require an external power source. The wet electro-material spontaneously generates an electrical effect by utilizing the humidity of the wound environment. At the same time, the electroactive bacteria in the wet electro-material produce hydrogen sulfide through metabolism to mediate gas therapy, effectively controlling wound inflammation and accelerating wound healing. This solves the problem that existing bioelectronic devices rely on rigid power sources and lack dynamic response capabilities.

[0005] In order to achieve the above-mentioned objectives of the present invention, the following technical solution is adopted: The first aspect of this invention provides a method for preparing a wet-electric material, the method comprising the following steps: (a) PEDOT:PSS was added to a solution of gelatin and alginate and mixed to obtain a MEG hydrogel solution; (b) Add Ag nanoparticles and Shewanella to a solution of gelatin and alginate and mix well to obtain a BFC hydrogel solution; (c) The MEG hydrogel solution and the BFC hydrogel solution are subjected to layered ionic crosslinking gelation using calcium chloride solution, followed by washing and freeze drying to obtain the wet electrical material.

[0006] Preferably, in step (a), the concentration of gelatin in the MEG hydrogel solution is 25~40 mg / mL, and the concentration of alginate is 8~12 mg / mL.

[0007] Preferably, in steps (a) and (b), the alginate is sodium alginate, potassium alginate, or ammonium alginate.

[0008] Preferably, in step (a), the concentration of PEDOT:PSS in the MEG hydrogel solution is 40~60 mg / mL.

[0009] Preferably, in step (b), the concentration of Ag nanoparticles in the BFC hydrogel solution is 8~12 mg / mL, and the concentration of Shewanella is (1~10)×10⁻⁶. 7 The concentration of CFU / mL, gelatin is 25-40 mg / mL, and alginate is 8-12 mg / mL.

[0010] Preferably, in step (b), the Shewanella is Lake Oneida Shewanella MR-1 ( Shewanella oneidensis MR-1).

[0011] Preferably, in step (c), the concentration of the calcium chloride solution is 20~40 mg / mL.

[0012] Preferably, in step (c), the temperature for ion crosslinking gelation is 3~6℃ and the time is 15~40min.

[0013] A second aspect of the present invention provides a wet electrical material prepared by the above-described preparation method.

[0014] A third aspect of the present invention provides the application of the wet electrochemical material prepared by the above-described preparation method in the preparation of wound healing dressings.

[0015] Compared with the prior art, the beneficial effects of the present invention include at least the following: This invention's wet-electric material triggers electrical stimulation therapy in response to the moist wound environment, exhibiting self-powered and dynamic response characteristics. The MEG and BFC loaded in this wet-electric material provide an electrical signal transmission network while releasing hydrogen sulfide, synergistically inducing electrical stimulation therapy and gas therapy. In the moist wound environment, MEG can release electrical stimulation, accelerating the migration of fibroblasts and keratinocytes to the wound site, promoting wound closure, and preventing bacterial infection; it also promotes angiogenesis and nerve regeneration, restoring skin function. Furthermore, the hydrogen sulfide produced by BFC promotes the transformation of macrophages from a pro-inflammatory phenotype to an anti-inflammatory phenotype, alleviating wound inflammation; hydrogen sulfide activates adipose tissue browning, secreting growth factors; and hydrogen sulfide enters the brain via blood circulation through the wound's blood vessels, relieving anxiety caused by the wound. Attached Figure Description

[0016] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the accompanying drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. In all the drawings, similar elements or parts are generally identified by similar reference numerals. In the drawings, the elements or parts are not necessarily drawn to scale.

[0017] Figure 1 This is a scanning electron microscope image of the wet electrochemical material in Experimental Example 1 of this invention; Figure 2 This is a scan of the elemental distribution surface of the energy dispersive spectroscopy of the wet electrochemical material in Experimental Example 1 of this invention; Figure 3 The results of the wet electrical performance test of the wet electrical material in Experiment Example 1 of this invention; Figure 4 This is the result of the study on the in vitro effects of different wet electrochemical materials on the proliferation of three types of cells in Experimental Example 2 of this invention; Figure 5 This is the result of the study on the in vitro effect of different wet electrochemical materials on fibroblast migration in Experimental Example 2 of the present invention; Figure 6 The results of animal experiments in Experiment Example 2 of this invention on the promotion of wound healing by different wet electrostatic materials; Figure 7The results of flow cytometry analysis in Experimental Example 2 of this invention are the effects of different wet electrochemical materials on macrophages. Figure 8 The experimental results of VEGFA in Experimental Example 2 of this invention show the effects of different wet electrostatic materials on blood vessels, nerves, and fat. Figure 9 The experimental results of CGRP in Experimental Example 2 of this invention show the effects of different wet electrostatic materials on blood vessels, nerves, and fat. Figure 10 Experimental results of UCP-1 in Experimental Example 2 of this invention regarding the effects of different wet electrostatic materials on blood vessels, nerves, and fat; Figure 11 The results of the open field experiment on the effect of different wet electrostatic materials on the anxiety behavior of mice in Experiment Example 2 of this invention. Detailed Implementation

[0018] The embodiments of the technical solution of the present invention will be described in detail below with reference to the examples. The following embodiments are only used to illustrate the technical solution of the present invention more clearly, and are therefore only examples, and should not be used to limit the scope of protection of the present invention.

[0019] It should be noted that, unless otherwise stated, the technical or scientific terms used in this application should have the ordinary meaning as understood by one of ordinary skill in the art to which this invention pertains.

[0020] This invention provides a method for preparing a wet-electric material, the method comprising the following steps: (a) PEDOT:PSS was added to a solution of gelatin and alginate and mixed to obtain a MEG hydrogel solution; (b) Add Ag nanoparticles and Shewanella to a solution of gelatin and alginate and mix well to obtain a BFC hydrogel solution; (c) The MEG hydrogel solution and the BFC hydrogel solution are subjected to layered ionic crosslinking gelation using calcium chloride solution, followed by washing and freeze drying to obtain the wet electrical material.

[0021] The wet electrochemical material of this invention is used to enhance wound electrical stimulation / gas therapy. Specifically, a high-performance hygroscopic and power-generating MEG hydrogel is prepared via a calcium chloride crosslinking method; Ag nanoparticles and Shewanella bacteria are in situ loaded onto gelatin and sodium alginate hydrogels to synthesize BFC hydrogels; the electroactivity of Shewanella bacteria is utilized to enhance the electrochemical properties of Ag nanoparticles and Shewanella bacteria. + The material is reduced to Ag nanoparticles on the bacterial surface, thereby enhancing the metabolic activity of the bacteria. When the wet electrochemical material is applied to the wound surface, the MEG hydrogel generates electrical signals under the stimulation of the moist environment on the body surface. The electroactive bacteria in the BFC hydrogel metabolize and release hydrogen sulfide, which accelerates wound healing through electrical stimulation and gas therapy.

[0022] In one embodiment, in step (a), the concentration of gelatin in the MEG hydrogel solution is 25-40 mg / mL, and the concentration of alginate is 8-12 mg / mL.

[0023] In one embodiment, in steps (a) and (b), the alginate is sodium alginate, potassium alginate, or ammonium alginate.

[0024] In one embodiment, in step (a), the concentration of PEDOT:PSS in the MEG hydrogel solution is 40~60 mg / mL.

[0025] In one embodiment, in step (b), the concentration of Ag nanoparticles in the BFC hydrogel solution is 8-12 mg / mL, and the concentration of Shewanella is (1-10) × 10⁻⁶. 7 The concentration of CFU / mL, gelatin is 25-40 mg / mL, and alginate is 8-12 mg / mL.

[0026] The source of Ag nanoparticles is not strictly limited in this invention, and commercially available Ag nanoparticles in the art can be used.

[0027] In one embodiment, in step (b), the Shewanella is Lake Oneida Shewanella MR-1.

[0028] In one embodiment, in step (c), the concentration of the calcium chloride solution is 20-40 mg / mL.

[0029] In one embodiment, in step (c), the temperature for ion crosslinking gelation is 3~6°C and the time is 15~40 min.

[0030] Another embodiment of the present invention provides a wet electrostatic material prepared by the above preparation method.

[0031] Another embodiment of the present invention provides the application of the wet electrochemical material prepared by the above preparation method in the preparation of wound healing dressings.

[0032] The technical solution of the present invention will be further described in detail below through specific embodiments. Example 1

[0033] This embodiment describes a method for preparing a wet-electric material, which includes the following steps: (a) Add 100 mg of sodium alginate to 10 mL of deionized water and stir vigorously for 30 min until completely dissolved; weigh 300 mg of gelatin and add it to 10 mL of sodium alginate solution, keep the temperature at 50℃ and stir until completely dissolved to obtain a hydrogel solution; weigh 500 mg of PEDOT:PSS and add it to 10 mL of hydrogel solution, keep the temperature at 37℃ and stir evenly to obtain a MEG hydrogel solution; (b) Add 100 mg of sodium alginate to 8 mL of deionized water and stir vigorously for 30 min until completely dissolved; weigh 300 mg of gelatin and add it to 8 mL of sodium alginate solution, maintaining the temperature at 50℃, and stir until completely dissolved; weigh 100 mg of Ag nanoparticles (Jiangsu Xianfeng Nanomaterials Technology Co., Ltd.) and add them to 8 mL of gelatin-sodium alginate solution, maintaining the temperature at 37℃, and stir evenly, then add 2 mL of Shewanella Oneida MR-1 culture medium (1×10⁻⁶). 8 (cfu / mL), stir well to obtain BFC hydrogel solution; (c) Add half the volume of BFC hydrogel solution to the mold, cool at 4°C for 10 min, then add MEG hydrogel solution, cool at 4°C for 10 min, add 3% (w / v) calcium chloride solution, and react at 4°C for 20 min to form a stable BFC+MEG hydrogel (denoted as BFC+MEG).

[0034] Comparative Example 1 This comparative example illustrates a method for preparing a wet-electric material, which includes the following steps: (a) Add 100 mg of sodium alginate to 10 mL of deionized water and stir vigorously for 30 min until completely dissolved; weigh 300 mg of gelatin and add it to 10 mL of sodium alginate solution, keep the temperature at 50℃ and stir until completely dissolved to obtain a hydrogel solution; weigh 500 mg of PEDOT:PSS and add it to 10 mL of hydrogel solution, keep the temperature at 37℃ and stir evenly to obtain a MEG hydrogel solution; (b) Add the MEG hydrogel solution into the mold, cool at 4°C for 20 min, add 3% (w / v) calcium chloride solution, and react at 4°C for 20 min to form a stable MEG hydrogel (denoted as MEG).

[0035] Comparative Example 2 This comparative example illustrates a method for preparing a wet-electric material, which includes the following steps: (a) Add 100 mg of sodium alginate to 8 mL of deionized water and stir vigorously for 30 min until completely dissolved; weigh 300 mg of gelatin and add it to 8 mL of sodium alginate solution, maintaining the temperature at 50℃, and stir until completely dissolved; weigh 100 mg of Ag nanoparticles (Jiangsu Xianfeng Nanomaterials Technology Co., Ltd.) and add them to 8 mL of gelatin-sodium alginate solution, maintaining the temperature at 37℃, and stir evenly, then add 2 mL of Shewanella Oneida MR-1 culture medium (1×10⁻⁶). 8 (cfu / mL), stir well to obtain BFC hydrogel solution; (b) Add the BFC hydrogel solution into the mold, cool at 4°C for 20 min, add 3% (w / v) calcium chloride solution, and react at 4°C for 20 min to form a stable BFC hydrogel (denoted as BFC).

[0036] Experimental Example 1 This experimental example characterizes the structure and properties of the wet-electric material in Example 1: 1. Structural Characterization Scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS) were used respectively; the electron microscopy results are as follows: Figure 1 As shown, the detection results of the energy scattering spectrometer are as follows: Figure 2 As shown; Depend on Figure 1 , Figure 2 It can be seen that the wet electrochemical material is composed of two hydrogels with different pore sizes. The BFC hydrogel has a dense structure and a high Ag content, while the MEG hydrogel has a loose structure and a high S content. The Ca element is evenly distributed in both hydrogels.

[0037] 2. Performance Characterization The wet electrostatic properties of the material were studied. Due to the hygroscopic discharge properties of PEDOT:PSS in the environment, the output voltage capability of the material under different humidity conditions was measured using an electrochemical workstation. The material was placed in environments with relative humidity (RH) of 30%, 50%, 70%, 90%, and 100%. The working electrode and reference electrode were fixed on the upper and lower surfaces of the material, respectively, and the voltage signals across the material were continuously measured. The test results are as follows: Figure 3 As shown; Depend on Figure 3 It can be seen that the wet-electric material outputs different voltages according to changes in ambient humidity. This phenomenon proves that the prepared wet-electric material has a high wet-electric conversion capability and can utilize the moist environment of the wound site to output electrical signals for electrical stimulation therapy.

[0038] Experimental Example 2 This experimental example verifies the efficacy of the wet electrical materials prepared in Example 1 and Comparative Examples 1-2: 1. Research on the effects of wet electrochemical materials on cell proliferation: L929, HaCat, and HUVEC cells were seeded in 12-well plates and cultured for 24 hours (37 °C, 5% CO2). Then, lyophilized MEG, BFC, and BFC+MEG hydrogels were placed in Transwell chambers. After co-culturing for 1, 3, and 5 days, CCK8 assays were performed to determine cell viability. The results are shown below. Figure 4 As shown; Depend on Figure 4 It is known that MEG and BFC promote cell proliferation through electrical stimulation and gas therapy, respectively, and their combined action accelerates cell proliferation through synergistic effects.

[0039] 2. Research on the effects of wet electrochemical materials on cell migration: L929, HaCat, and HUVEC cells were seeded in 12-well plates and cultured for 48 hours (37 °C, 5% CO2). Then, 200 μm wide scratches were created in the wells, and lyophilized MEG, BFC, and BFC+MEG hydrogels were placed in Transwell chambers. After co-culturing for 0, 24, and 48 hours, the healing status of the cell wounds was recorded under a microscope. The results are shown below. Figure 5 As shown; Depend on Figure 5 It is known that MEG and BFC promote cell migration through electrical stimulation and gas therapy, respectively, and the cell wounds healed first after the two act together.

[0040] 3. Animal experiments demonstrating that wet electrostatic materials promote wound healing in diabetic mice: All groups of mice used five male C57BL / 6J mice approximately 8 weeks old. A 10% streptozotocin (STZ) solution was prepared using sodium citrate buffer (pH 4.5) and used immediately. Mice were intraperitoneally injected with STZ (100 mg / kg) for three consecutive days based on their body weight. One week later, blood was collected from the tail vein; a blood glucose level ≥16.7 mmol / L in three randomized measurements was considered a successful diabetes model. A full-thickness skin lesion of approximately 8 mm in diameter was made on the back of the mouse, and 20 μL of Staphylococcus aureus was inoculated into the lesion. S.aureus ) suspension (1×10 8 (cfu / mL). One day later, the commercially available wound dressing 3M™ Tegaderm™ was used as a control group. The experimental groups used the wet electrostatic material BFC+MEG lyophilized gel, the BFC group used BFC lyophilized gel, and the MEG group used MEG lyophilized gel. The observation period was 9 days, and photos were taken and recorded. The results are as follows: Figure 6 As shown; Depend on Figure 6It can be seen that the wound healing speed of the experimental group and other groups of mice was consistently greater than that of the control group, while the wound healing speed of other groups of mice was less than that of the experimental group but greater than that of the control group. This shows that the wet electrostatic material of the present invention can accelerate wound healing.

[0041] 4. Research on the effects of wet electrochemical materials on macrophages: Five diabetic mice with wound infection were used in both the control and experimental groups. On the seventh day after wound healing, three mice from each group were randomly selected, and skin tissue was collected from the wound site. Cells were collected after collagenase digestion, stained with CD11b, CD86, and CD206 antibodies, and finally analyzed by flow cytometry. The results are as follows: Figure 7 As shown; Depend on Figure 7 The results showed that the M2 / M1 cell ratio in the experimental group increased by 3.6 times compared with the control group. These results confirm that the wet electrochemical material promotes the transformation of macrophages into the anti-inflammatory (M2) type, thus exhibiting an anti-inflammatory effect.

[0042] 5. Effects of wet-electric materials on blood vessels, nerves, and fat: Five diabetic mice with wound infection were used in both the control and experimental groups. On the seventh day after wound healing, three mice from each group were randomly selected, and skin tissue was harvested from the wound site. The tissue was fixed in 10% paraformaldehyde fixative, routinely dehydrated and embedded, and immunofluorescent VEGFA, CGRP, and UCP-1 stained sections were prepared. The results are as follows: Figures 8-10 As shown; Depend on Figures 8-10 It is known that VEGFA is an angiogenic marker, CGRP marks sensory neurons, and UCP-1 is a marker of adipocyte browning. In the experimental group of mice, the fluorescence intensity of VEGFA, CGRP, and UCP-1 in the wound tissue was greater than that in the control group. The fluorescence intensity in other groups of mice was lower than that in the experimental group but higher than that in the control group. Therefore, it can be concluded that wet electrochemical materials can promote angiogenesis and nerve regeneration and adipocyte browning in wounds.

[0043] 6. Effects of wet electrical materials on anxiety behavior in mice: Five diabetic mice with wound infection were used in both the control and experimental groups. An open field test was performed on the mice on the seventh day after wound healing. The results are as follows: Figure 11 As shown; Depend on Figure 11 It was found that the experimental group mice spent a longer period of time in the central region of the open field compared to the control group mice. Anxiety caused the mice to tend to move along the periphery of the open field, and the increased time spent in the central region indicates that the mice's anxiety was alleviated. Therefore, it can be concluded that the wet electrostatic material has the effect of alleviating anxiety in mice.

[0044] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention, and they should all be covered within the scope of the claims and specification of the present invention.

Claims

1. A method for preparing a wet-electric material, characterized in that, The preparation method of the wet electrostatic material includes the following steps: (a) PEDOT:PSS was added to a solution of gelatin and alginate and mixed to obtain a MEG hydrogel solution; (b) Add Ag nanoparticles and Shewanella to a solution of gelatin and alginate and mix well to obtain a BFC hydrogel solution; (c) The MEG hydrogel solution and the BFC hydrogel solution are subjected to layered ionic crosslinking gelation using calcium chloride solution, followed by washing and freeze drying to obtain the wet electrical material.

2. The method for preparing the wet-electric material according to claim 1, characterized in that, In step (a), the concentration of gelatin in the MEG hydrogel solution is 25~40 mg / mL, and the concentration of alginate is 8~12 mg / mL.

3. The method for preparing the wet-electric material according to claim 1, characterized in that, In steps (a) and (b), the alginate is sodium alginate, potassium alginate, or ammonium alginate.

4. The method for preparing the wet-electric material according to claim 1, characterized in that, In step (a), the concentration of PEDOT:PSS in the MEG hydrogel solution is 40~60 mg / mL.

5. The method for preparing the wet-electric material according to claim 1, characterized in that, In step (b), the concentration of Ag nanoparticles in the BFC hydrogel solution is 8~12 mg / mL, and the concentration of Shewanella is (1~10)×10⁻⁶. 7 The concentration of CFU / mL, gelatin is 25-40 mg / mL, and alginate is 8-12 mg / mL.

6. The method for preparing the wet-electric material according to claim 1, characterized in that, In step (b), the Shewanella is Lake Oneida Shewanella MR-1.

7. The method for preparing the wet-electric material according to claim 1, characterized in that, In step (c), the concentration of the calcium chloride solution is 20~40 mg / mL.

8. The method for preparing the wet-electric material according to claim 1, characterized in that, In step (c), the temperature for ion crosslinking gelation is 3~6℃ and the time is 15~40min.

9. The wet electrical material prepared by any one of claims 1 to 8.

10. The use of the wet electrochemical material prepared by any one of claims 1 to 8 in the preparation of wound healing dressings.

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