Cell-mimic extracellular matrix anti-inflammatory and nerve repair-promoting conductive hydrogel, and preparation method and application thereof

Abstract

The application discloses a kind of extracellular matrix imitated anti-inflammatory nerve repair promoting conductive hydrogel and its preparation method and application, belong to high polymer material technical field.The preparation method of the conductive hydrogel includes the following steps: preparation MXene nanosheet dispersion liquid, polyphenol and magnesium ion are sequentially added into the MXene nanosheet dispersion liquid, and modified nanosheet solution is obtained;Modified chitosan and polyvinyl alcohol are mixed to obtain solution A;The modified nanosheet solution is mixed with phenylboronic acid modified gelatin after reaction to obtain solution B;Solution A and solution B are mixed and reacted after low-temperature refrigeration, and the conductive hydrogel is obtained after thawing.The hydrogel prepared by the application has excellent conductive performance, and the obtained conductive hydrogel has anti-inflammatory and nerve repair promoting functions.This bionics method not only improves the diversity and functionality of biomaterials, but also provides new possibilities for future tissue engineering and regenerative medicine.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of high polymer materials, in particular to a kind of extracellular matrix-like anti-inflammatory nerve repair promoting conductive hydrogel and its preparation method and application. BACKGROUND

[0002] Bionics thinking can well expand the diversity of biomaterials. It is well known that extracellular matrix (ECM) is a very complex fluid environment, which not only supports cell growth and differentiation as a three-dimensional scaffold, but also controls cell behavior by regulating dynamic mechanical and chemical signals and their microenvironment, and promotes tissue repair during injury. In recent years, hydrogel as a three-dimensional material with high water content has developed most rapidly in the field of biological tissue repair. Hydrogel has good biocompatibility and adjustable physical properties, making it an ideal choice to simulate natural ECM.

[0003] However, hydrogel itself is not conductive, which limits its use in some applications that require electrical activity. Conductive hydrogel can form an endogenous electric field, which has a significant promoting effect on angiogenesis, nerve repair and tissue healing, and therefore can be widely used in repairing electrically active tissues such as wounds, myocardium and nerves. Combining conductive hydrogel with hydrophilic matrix and conductive medium organically is a new type of composite hydrogel with good processability, high flexibility and excellent electrochemical performance, which has broad application prospects in many fields such as electronic skin, biosensor, supercapacitor, flexible wearable electronic device, and is an ideal material for future flexible electronic devices. However, most traditional chemical cross-linked hydrogels have the problem of insufficient conductivity, which is also the key to limit the application of hydrogels. Therefore, it is of great significance to provide a hydrogel preparation scheme with excellent conductivity. SUMMARY

[0004] The purpose of the present application is to provide an extracellular matrix-like anti-inflammatory nerve repair promoting conductive hydrogel and its preparation method and application, in order to solve the problems existing in the prior art. The hydrogel prepared by the present application has similar components, structure, mechanical elasticity and excellent conductivity to extracellular matrix, and the obtained conductive hydrogel has anti-inflammatory and nerve repair promoting functions.

[0005] In order to solve the above problems, the present application provides the following scheme:

[0006] Technical Solution 1: A method for preparing a conductive hydrogel, comprising the following steps: preparing an MXene nanosheet dispersion; sequentially adding polyphenols and magnesium ions to the MXene nanosheet dispersion to react and obtain a modified nanosheet solution; mixing the modified chitosan with polyvinyl alcohol to obtain solution A; mixing the modified nanosheet solution with phenylboronic acid-modified gelatin to react and obtain solution B; mixing solution A and solution B and then refrigerating at low temperature, and obtaining the conductive hydrogel after thawing.

[0007] Further, the preparation of the modified nanosheet solution includes the following steps: adding LiF to hydrochloric acid solution, adding Ti3AlC2 and mixing; collecting the mixed precipitate, centrifuging and washing with deionized water until the pH of the supernatant reaches 6; collecting the precipitate obtained after centrifugation, ultrasonically dispersing it in an Ar environment, centrifuging it, collecting the solution, and obtaining an MXene nanosheet dispersion; adding gallic acid or tannic acid solution dropwise to the MXene nanosheet dispersion and mixing it, then adding MgCl2·6H2O solution dropwise, mixing, centrifuging and washing to obtain the modified nanosheet solution.

[0008] Specifically, the preparation of the modified nanosheet solution includes the following steps: 1-3 g of LiF is added to 20-60 mL of 9-10 M hydrochloric acid solution and stirred thoroughly; then 1-3 g of Ti3AlC2 is slowly added and stirred at 35-45°C for 24-28 hours; the precipitate is collected and centrifuged at 4500 rpm for 5-10 minutes with deionized water until the pH of the supernatant reaches 6; then the precipitate is collected, 50-100 mL of deionized water is added, and the mixture is ultrasonically dispersed in an Ar environment for 1-1.5 hours, centrifuged, and the solution is collected. 10-50 mg of gallic acid or tannic acid is dissolved in Tris at pH 8.5 and then slowly added dropwise to a 10 mg / mL MXene nanosheet dispersion and stirred in the dark for 2-4 hours. Then, 50-100 mg of MgCl2·6H2O was added to 1-5 mL of Tris solution and added dropwise to the previous solution for reaction. The mixture was stirred continuously, and finally centrifuged at 8000-10000 rpm for 3-10 minutes. The precipitate was washed with deionized water and collected to obtain the modified nanosheet solution.

[0009] Further, the preparation method of the modified chitosan includes the following steps: dissolving dihydrocaffeic acid and adding it to chitosan solution to obtain mixed solution 1; dissolving 1-ethyl-(3-dimethylaminopropyl)carbodiimide and N-hydroxythiosuccinimide in a mixed solution of ethanol and deionized water to obtain mixed solution 2; adding mixed solution 2 to mixed solution 1 for reaction; introducing Ar and reacting to obtain product solution; dialyzing first in acidified deionized water, then dialysis in deionized water, and finally freeze-drying.

[0010] Specifically, the preparation method of the modified chitosan includes the following steps: 0.5-0.8 g of chitosan is dissolved in deionized water, the pH is adjusted to 5.0-6.0 with HCl, and the mixture is stirred in a water bath at 60-80°C for 20-30 min; 255-455 mg of dihydrocaffeic acid is dissolved in 5.0-8.0 mL of deionized water, and after thorough stirring and dissolution, it is added to the chitosan solution; 240-300 mg of 1-ethyl-(3-dimethylaminopropyl)carbodiimide and 150-200 mg of N-hydroxythiosuccinimide are dissolved in a 1:1-1:2 mixture of ethanol and deionized water, and the mixture is added to the mixture. Ar is introduced and the mixture is reacted at room temperature for 12 h; the product solution is dialyzed in acidified deionized water for 2-3 days, then dialyzed in deionized water for another 0.5 days, and finally freeze-dried in a freeze dryer.

[0011] Furthermore, the preparation method of the modified gelatin includes the following steps:

[0012] Add 4–8 g of gelatin to 400–800 mL of deionized water and stir thoroughly at 60–80 °C. Add 7–10 g of NHS and 1.5–3 g of EDS powder to the above solution and stir evenly. Adjust the pH to 5 with 1–5 M hydrochloric acid solution. Then add 3–6 g of PBA powder and react in a water bath at 37–45 °C for 48 h. Dialyze the resulting solution in deionized water for 3 days and then freeze-dry it.

[0013] Furthermore, the preparation method of solution A includes: dissolving the polyvinyl alcohol powder, adding the modified chitosan to the dissolved polyvinyl alcohol solution, and mixing and reacting to obtain solution A.

[0014] The specific preparation method of solution A is as follows: 1-3g of PVA powder is added to 10-30mL of deionized water and dissolved at 95-100℃. 0.1-0.3g of CSDA is added to the dissolved PVA solution and the mixture is stirred and reacted for 1-2 hours to obtain solution A.

[0015] Further, the preparation method of solution B includes: preparing a 12.5 wt% solution of phenylboronic acid-modified gelatin, and adding the modified nanosheet solution to it to obtain solution B.

[0016] The specific preparation method of solution B is as follows: prepare a 12.5 wt% solution of phenylboronic acid-modified gelatin, and add 20-60 μL of the modified nanosheet solution at a concentration of 10 mg / mL to obtain solution B.

[0017] Furthermore, solution A and solution B are mixed and reacted in a volume ratio of 1:1.

[0018] Furthermore, the low-temperature refrigeration is refrigerating at -80°C for 12 to 48 hours.

[0019] Technical Solution 2: The conductive hydrogel prepared by the aforementioned preparation method.

[0020] Technical Solution 3: The application of the conductive hydrogel in the preparation of hydrogels with extracellular matrix-like and anti-inflammatory effects.

[0021] Technical Solution 4: The application of the conductive hydrogel in the preparation of hydrogels that promote nerve cell proliferation and repair.

[0022] The present invention discloses the following technical effects:

[0023] This invention provides an extracellular matrix-inspired anti-inflammatory, nerve-repairing, and conductive hydrogel, its preparation method, and its biological applications. The invention first prepares MXene nanosheets, then sequentially adds polyphenols and magnesium ions to the MXene nanosheet solution and stirs; after thorough mixing and washing, the modified nanosheets are obtained, which possess conductive, anti-inflammatory, and nerve-repairing functions. The preparation method includes mixing modified chitosan with polyvinyl alcohol to obtain solution A, and mixing the novel nanosheets with modified gelatin to obtain solution B. Solutions A and B are mixed and reacted in a certain proportion, then refrigerated at low temperature for a period of time to obtain the conductive hydrogel.

[0024] The hydrogel synthesized in this invention uses widely available, biocompatible, and non-toxic raw materials, which is beneficial for product commercialization. The hydrogel provided by this invention has an easy and rapid gelation process. The conductive hydrogel provided by this invention mimics the extracellular matrix, which is beneficial for tissue repair. The hydrogel prepared by this invention has similar composition, structure, mechanical elasticity, and excellent conductivity to the extracellular matrix, and the obtained conductive hydrogel also has anti-inflammatory and nerve-repairing functions. The method provided by this invention not only improves the diversity and functionality of biomaterials but also provides new possibilities for future tissue engineering and regenerative medicine. Attached Figure Description

[0025] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0026] Figure 1 The image shows a transmission electron microscope (TEM) image of the modified nanosheet solution prepared in Example 1, in which magnesium ions are uniformly distributed on the MXene nanosheets. In the image, A is the TEM image of the modified nanosheet, B is the surface O element distribution, C is the Ti element distribution, and D is the Mg element distribution.

[0027] Figure 2 Photograph of the conductive hydrogel in Example 1;

[0028] Figure 3 The free radical scavenging ability of the hydrogel prepared in Example 1;

[0029] Figure 4 Immunofluorescence image of the hydrogel prepared in Example 1 promoting nerve cell proliferation and development;

[0030] Figure 5 The relative fluorescence area of ​​the hydrogel prepared in Example 1;

[0031] Figure 6 The conductivity of the hydrogel prepared in Example 2 is such that it has a conductivity that matches that of nerve cells. Detailed Implementation

[0032] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0033] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0034] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0035] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be readily apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0036] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0037] EGCG can be represented as tannic acid or gallic acid.

[0038] Example 1

[0039] 1. Preparation of conductive hydrogels

[0040] (1) Preparation of modified nanosheet solution: 1 g LiF was added to 20 mL of 9 M hydrochloric acid solution and stirred thoroughly. Then 1 g Ti3AlC2 was slowly added and stirred at 45 °C for 26 h. The precipitate was collected and centrifuged at 4500 rpm for 5 min with deionized water until the pH of the supernatant reached 6. The centrifuged precipitate was then collected and added to 60 mL of deionized water and ultrasonically dispersed in Ar environment for 1 h. The solution was then centrifuged and collected, which is the MXene nanosheet dispersion. 10 mg of tannic acid (EGCG) was dissolved in Tris buffer at pH 8.5 and then slowly added dropwise to the 10 mg / mL MXene nanosheet dispersion. After stirring in the dark for 2 h, MgCl2·6H2O solution was added and the reaction was continued. Finally, the mixture was centrifuged at 8000 rpm for 5 min and washed with deionized water to collect the precipitate MXene@EGCG-Mg (modified nanosheet solution). The MgCl2·6H2O solution was obtained by dissolving 50 mg of MgCl2·6H2O in 2 mL of Tris solution.

[0041] (2) Preparation of catechol-modified chitosan (CHIC): 0.5 g chitosan was dissolved in deionized water, and the pH was adjusted to 5.0 with HCl. The mixture was stirred in a 60°C water bath for 20 min. Then, 260 mg of HCA (dihydrocaffeic acid) was dissolved in 5.0 mL of deionized water and added to the chitosan solution after thorough stirring. 240 mg of 1-ethyl-(3-dimethylaminopropyl)carbodiimide (EDC) and 150 mg of N-hydroxythiosuccinimide (NHS) were dissolved in a 1:1 mixture of ethanol and deionized water in 50.0 mL. This mixture was then added to the solution, Ar was introduced, and the mixture was reacted at room temperature for 12 h. The solution was dialyzed in acidified deionized water for 2 days, then dialyzed again in deionized water for 0.5 days, and finally freeze-dried to obtain a yellow solid.

[0042] (3) Phenylboronic acid modified gelatin (GPBA): 4 g of gelatin was added to 400 mL of deionized water and stirred thoroughly at 70 °C. 7 g of NHS (N-hydroxythiosuccinimide) and 2 g of EDS (N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride) powder were added to the above solution and stirred until homogeneous. The pH was then adjusted to 5 with hydrochloric acid solution. Then, 3.5 g of PBA powder was added and the mixture was stirred in a water bath at 37 °C for 48 h. The resulting solution was then dialyzed in deionized water for 3 days and then freeze-dried.

[0043] (4) Dissolve 1g of PVA (polyvinyl alcohol) powder in 10mL of deionized water at 95℃. Add 0.1g of catechol-modified chitosan to the dissolved PVA solution and stir for 1h to obtain solution A. Then, prepare a 12.5wt% GPBA solution and add 20μL of 10mg / mL MXene@EGCG-Mg to obtain solution B. Mix solutions A and B at a volume ratio of 1:1 and refrigerate at -80℃ for 12h. Thaw at room temperature to obtain conductive hydrogel.

[0044] 2. Determination of the properties of conductive hydrogels

[0045] (1) The electrochemical performance of the hydrogel patch was tested using a dual-probe method on an electrochemical workstation (CHI660C). The hydrogel was sandwiched between two copper plates for measurement. During the impedance measurement, the voltage was kept constant at 50mV, and the frequency range was 10. 6 -10 -1 Hz.

[0046] (2) The antioxidant properties of each component of the hydrogel were tested using the DPPH free radical scavenging method. First, the DPPH solution was prepared, protected from light, and stored. Then, the hydrogel was prepared to a concentration of 2.0 mg / mL. After mixing the DPPH solution with 100 μL of the sample solution, the mixture was placed in the dark for 30–120 min. Finally, the solution was scanned using a microplate reader.

[0047] (3) After culturing PC12 cells for 7 days, wash them three times with PBS, fix them with 4% paraformaldehyde for 20 minutes, and infiltrate them with 0.3% Triton X-100 for 5 minutes. After blocking with 1% BSA for 1 hour, incubate them overnight at 4°C with diluted primary antibody (Tuj1), and then incubate them with secondary antibody at room temperature for 1 hour. Wash the cells three times with PBS and stain them with DAPI for 5 minutes. Finally, examine the cells using a laser scanning confocal microscope.

[0048] 3. Experimental Results

[0049] Transmission electron microscopy image of the prepared MXene@EGCG-Mg nanosheets is shown below. Figure 1(200nm) Magnesium ions are visible uniformly distributed on the MXene nanosheets; the conductive hydrogel gelation image is shown below. Figure 2 The free radical scavenging ability of the prepared hydrogel is shown in the figure. Figure 3 The immunofluorescence image of the prepared hydrogel promoting nerve cell proliferation and development is shown in the figure. Figure 4 The PCP group represents a hydrogel with a nanosheet concentration of 0 mg / mL, and the PCPM2 group represents a hydrogel with a nanosheet concentration of 0.4 mg / mL; the relative fluorescence area is shown in [reference needed]. Figure 5 The PCP group represents a hydrogel with a nanosheet concentration of 0 mg / mL. In summary, the conductive hydrogel prepared in this invention exhibits high conductivity and free radical scavenging ability, and can promote nerve cell proliferation.

[0050] Example 2

[0051] 1. Preparation of conductive hydrogels

[0052] (1) First, add 2g of LiF to 40mL of 10M hydrochloric acid solution and stir thoroughly. Then, slowly add 2g of Ti3AlC2 and stir at 35℃ for 28h. Collect the precipitate and centrifuge at 4500 rpm for 8min with deionized water until the pH of the supernatant reaches 6. Then, collect the precipitate, add 100mL of deionized water, sonicate for 1.5h in Ar environment, and centrifuge to collect the solution. Dissolve 50mg of gallic acid (EGCG) in Tris buffer at pH=8.5, and then slowly add it dropwise to 10mg / mL MXene nanosheet dispersion and stir in the dark for 2h. Then, add 60mg of MgCl2·6H2O to 2mL of Tris solution and dropwise to the previous solution, and continue stirring the reaction. Finally, centrifuge at 10000 rpm for 5min and wash with deionized water to collect the precipitate MXene@EGCG-Mg (modified nanosheet solution).

[0053] (2) 0.8 g of chitosan was dissolved in deionized water, and the pH was adjusted to 5.0 with HCl. The solution was stirred in an 80°C water bath for 30 min. Then, 455 mg of HCA was dissolved in 8.0 mL of deionized water and added to the chitosan solution after thorough stirring. 300 mg of 1-ethyl-(3-dimethylaminopropyl)carbodiimide (EDC) and 200 mg of N-hydroxythiosuccinimide (NHS) were dissolved in a 1:1 mixture of ethanol and deionized water in 50.0 mL. This mixture was then added to the solution, Ar was introduced, and the mixture was reacted at room temperature for 12 h. The solution was dialyzed in acidified deionized water for 3 days, then dialyzed in deionized water for another 0.5 days, and finally freeze-dried to obtain a yellow solid.

[0054] (3) Add 6g of gelatin to 600mL of deionized water and stir thoroughly at 80℃. Add 10g of NHS and 3g of EDS powder to the above solution and stir evenly. Adjust the pH to 5 with hydrochloric acid solution. Then add 5g of PBA powder and react in a water bath at 45℃ for 48h. After that, dialysis the resulting solution in deionized water for 3 days and then freeze-dry it.

[0055] (4) 2g of PVA powder was dissolved in 20mL of deionized water at 100℃. 0.2g of modified chitosan was added to the dissolved PVA solution and stirred for 2h to obtain solution A. Then, GPBA was prepared into a 12.5wt% solution, and 10mg / mL of MXene@EGCG-Mg was added to obtain solution B. Solutions A and B were mixed at a volume ratio of 1:1 and reacted. The mixture was then refrigerated at -80℃ for 48h and thawed at room temperature to obtain a conductive hydrogel.

[0056] 2. Determination of the properties of conductive hydrogels

[0057] (1) The electrochemical performance of the hydrogel patch was tested using a dual-probe method on an electrochemical workstation (CHI660C). The hydrogel was sandwiched between two copper plates for measurement. During the impedance measurement, the voltage was kept constant at 50mV, and the frequency range was 10. 6 -10 -1 Hz.

[0058] (2) The antioxidant properties of each component of the hydrogel were tested using the DPPH free radical scavenging method. First, the DPPH solution was prepared, protected from light, and stored. Then, the hydrogel was prepared to a concentration of 2.0 mg / mL. After mixing the DPPH solution with 100 μL of the sample solution, the mixture was placed in the dark for 30–120 min. Finally, the solution was scanned using a microplate reader.

[0059] After culturing PC12 cells for 7 days, they were washed three times with PBS, fixed with 4% paraformaldehyde for 20 minutes, and infiltrated with 0.3% Triton X-100 for 5 minutes. After blocking with 1% BSA for 1 hour, they were incubated overnight at 4°C with diluted primary antibody (Tuj1), followed by incubation with secondary antibody at room temperature for 1 hour. The cells were washed three times with PBS and stained with DAPI for 5 minutes. Finally, the cells were examined using a laser scanning confocal microscope.

[0060] 3. Experimental Results

[0061] Figure 6The conductivity of the hydrogels prepared in this invention is shown to be comparable to that of nerve cells. Specifically, the PCP group represents hydrogels with a nanosheet concentration of 0 mg / mL, the PCPM1 group represents hydrogels with a nanosheet concentration of 0.2 mg / mL, the PCPM2 group represents hydrogels with a nanosheet concentration of 0.4 mg / mL, and the PCPM3 group represents hydrogels with a nanosheet concentration of 0.6 mg / mL. Transmission electron microscopy images of the prepared modified nanosheet solutions are shown below. Figure 1 A(200nm); Photograph of conductive hydrogel formation is shown below. Figure 2 The free radical scavenging ability of the prepared hydrogel is shown in the figure. Figure 3 The immunofluorescence image of the prepared hydrogel promoting nerve cell proliferation and development is shown in the figure. Figure 4 The relative fluorescence area is shown in the figure. Figure 5 .

[0062] In summary, the conductive hydrogel prepared by this invention has high conductivity and the ability to scavenge free radicals, can exert anti-inflammatory effects, and can promote nerve cell proliferation and promote nerve repair.

[0063] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. The application of conductive hydrogels in the preparation of hydrogels that promote nerve cell proliferation and repair, characterized in that, The method for preparing the conductive hydrogel includes the following steps: To prepare an MXene nanosheet dispersion, polyphenols and magnesium ions were added sequentially to the MXene nanosheet dispersion to obtain a modified nanosheet solution; the modified chitosan was then mixed with polyvinyl alcohol to react and obtain solution A. Solution B is obtained by mixing and reacting the modified nanosheet solution with phenylboronic acid-modified gelatin. Solution A and solution B were mixed and reacted, then refrigerated at low temperature, and the conductive hydrogel was obtained after thawing.

2. The application according to claim 1, characterized in that, The preparation of the modified nanosheet solution includes the following steps: adding LiF to hydrochloric acid solution, adding Ti3AlC2 and mixing; collecting the precipitate after mixing, centrifuging and washing with deionized water until the pH of the supernatant reaches 6; collecting the precipitate obtained after centrifugation, ultrasonically dispersing it in an Ar environment, centrifuging it, collecting the solution, and obtaining an MXene nanosheet dispersion; adding gallic acid or tannic acid solution dropwise to the MXene nanosheet dispersion and mixing, then adding MgCl2·6H2O solution dropwise, mixing, centrifuging and washing to obtain the modified nanosheet solution.

3. The application according to claim 1, characterized in that, The method for preparing the modified chitosan includes the following steps: dissolving dihydrocaffeic acid and adding it to a chitosan solution to obtain a mixed solution 1; dissolving 1-ethyl-(3-dimethylaminopropyl)carbodiimide and N-hydroxythiosuccinimide in a mixed solution of ethanol and deionized water to obtain a mixed solution 2; adding the mixed solution 2 to the mixed solution 1; introducing Ar and reacting to obtain a product solution; dialyzing first in acidified deionized water; then dialysis in deionized water; and finally freeze-drying.

4. The application according to claim 1, characterized in that, The preparation method of solution A includes: dissolving the polyvinyl alcohol powder, adding the modified chitosan to the dissolved polyvinyl alcohol solution, and mixing to obtain solution A.

5. The application according to claim 1, characterized in that, The preparation method of solution B includes: preparing a 12.5 wt% solution of phenylboronic acid-modified gelatin, and adding modified nanosheets to it to obtain solution B.

6. The application according to claim 1, characterized in that, Solution A and solution B are mixed and reacted in a volume ratio of 1:

1.

7. The application according to claim 1, characterized in that, The low-temperature refrigeration refers to placing the container at -80℃ for 12 to 48 hours.

8. The application of conductive hydrogels in the preparation of hydrogels with extracellular matrix-like and anti-inflammatory effects, characterized in that, The method for preparing the conductive hydrogel includes the following steps: To prepare an MXene nanosheet dispersion, polyphenols and magnesium ions were added sequentially to the MXene nanosheet dispersion to obtain a modified nanosheet solution; the modified chitosan was then mixed with polyvinyl alcohol to react and obtain solution A. Solution B is obtained by mixing and reacting the modified nanosheet solution with phenylboronic acid-modified gelatin. Solution A and solution B were mixed and reacted, then refrigerated at low temperature, and the conductive hydrogel was obtained after thawing.

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