A pro-healing high-adhesion enzymatic hydrogel for vascular intima and a preparation method thereof

Abstract

The application discloses a kind of high adhesion enzymatic hydrogel for promoting vascular intimal repair and a preparation method thereof, and belongs to the technical field of hydrogel preparation.The preparation method is as follows: S1, using methacrylic anhydride to modify the double bond of hyaluronic acid, then crosslinking with dopamine to obtain dopamine-modified double-bonded hyaluronic acid;S2, after removing oxygen from the hyaluronic acid solution, add a crosslinking agent and tyramine hydrochloride, and react in an acidic environment to obtain tyramine-modified hyaluronic acid;S3, mix the dopamine-modified double-bonded hyaluronic acid, tyramine-modified hyaluronic acid and deionized water to obtain a hydrogel precursor solution;S4, then use enzymatic reaction to obtain the hydrogel.The method designed by the application is simple, does not require additional light curing equipment, and the constructed hydrogel is similar to the extracellular matrix.The hydrogel can be coated into different shapes to adapt to different environments.The hydrogel is formed by enzymatic reaction, which is rapid, well adhered to the tissue, biocompatible, and suitable for application in vascular repair.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of hydrogel preparation, and particularly relates to a high-adhesion enzymatic hydrogel for promoting vascular intima repair and a preparation method thereof. BACKGROUND

[0002] Atherosclerosis is the culprit of cardiovascular disease. Vascular stent implantation is one of the important means for treating cardiovascular disease in clinic, which can improve the blood vessel occlusion by expanding the lesion site. However, it is accompanied by the risk of complications such as restenosis, late thrombosis and late neovascular atherosclerosis, which may require reoperation to bring more risks to patients, and the stent is non-degradable and needs to exist in the human body for a long time after implantation.

[0003] Hydrogel is a material similar to the extracellular matrix, which is often used for skin wound repair and the like, and the gelation time and adhesion capacity of the hydrogel can be adjusted by changing the gel components. Such repair-promoting hydrogel is often applied to the static surface of the skin and the like, and is easy to fall off when subjected to fluid flushing, and the use of a mold for production results in that the hydrogel cannot completely match the vascular intima. At the same time, the current hydrogel gelation usually needs to add a photoinitiator and irradiate ultraviolet light for curing, and the operation is more complicated. Moreover, in the vascular intima environment, there is a high-speed blood flow flushing, and a large number of platelets exist therein, and the anti-platelet adhesion performance of the hydrogel is also a great challenge. SUMMARY

[0004] In view of the above problems in the prior art, the application provides a high-adhesion enzymatic hydrogel for promoting vascular intima repair and a preparation method thereof. The method designed by the application is simple in process, does not need to add a light curing device, and the constructed hydrogel is similar to the extracellular matrix. The hydrogel can be coated into different shapes to adapt to different environments. The hydrogel is rapidly gelled by enzymatic reaction, has good adhesion to tissues, good biocompatibility, and is suitable for application in vascular repair after atherectomy.

[0005] To achieve the above object, the technical scheme adopted by the application to solve the technical problem is as follows:

[0006] The application aims to provide a preparation method of a high-adhesion enzymatic hydrogel for promoting vascular intima repair, comprising the following steps:

[0007] S1, dopamine-modified double-bonded hyaluronic acid (HMD)

[0008] The hyaluronic acid is double-bonded and modified by using methacrylic anhydride, and then crosslinked with dopamine to obtain dopamine-modified double-bonded hyaluronic acid;

[0009] S2, tyrosine-modified hyaluronic acid (HT)

[0010] The hyaluronic acid solution is added with a cross-linking agent and tyramine hydrochloride after oxygen removal, and reacted in an acidic environment for 4-8 h to prepare tyramine-modified hyaluronic acid;

[0011] S3, mixing the dopamine-modified double-bonded hyaluronic acid solution and the tyramine-modified hyaluronic acid solution to obtain a hydrogel precursor liquid with a mass concentration of 2-3 wt% of dopamine-modified double-bonded hyaluronic acid and tyramine-modified hyaluronic acid;

[0012] S4, using an enzyme reaction to prepare a high-adhesion enzyme hydrogel for promoting vascular intimal repair.

[0013] Further, the process of modifying hyaluronic acid by double-bonding with methacrylic anhydride is as follows:

[0014] Methacrylic anhydride is added to the hyaluronic acid solution, the pH is adjusted to 8-9, and the reaction is carried out for 2-3 h. Ethanol is added for precipitation, and the precipitate is washed with alcohol and then dissolved in deionized water. The solution is dialyzed and freeze-dried to obtain the product.

[0015] The mass concentration of hyaluronic acid is 1-1.5 wt%, and the mass concentration of methacrylic anhydride is 4.5-5 wt%.

[0016] Further, the process of modifying hyaluronic acid by double-bonding with methacrylic anhydride is as follows:

[0017] The double-bonded modified hyaluronic acid is dissolved in MES buffer to obtain a solution with a mass concentration of 1-1.5 wt%. A cross-linking agent and dopamine (DOPA) are added, the pH is adjusted to 5-6 after oxygen removal, and the reaction is carried out for 4-8 h. The pH is then adjusted to be acidic, and the product is obtained by dialysis and freeze-drying.

[0018] The concentration of the MES buffer is 100-120 mM, and the mass concentration of dopamine is 0.3-0.5 wt%.

[0019] Further, the cross-linking agent used is NHS / EDC, wherein the mass concentration of NHS is 0.2-0.3 wt%, and the mass concentration of EDC is 0.3-0.4 wt%.

[0020] Further, the mass concentration of hyaluronic acid in S2 is 0.5-1.5 wt%, and the concentration of tyramine hydrochloride is 1.2-1.5 mM.

[0021] Further, the cross-linking agent used in S2 is NHS / EDC, wherein the mass concentration of NHS is 0.2-0.3 wt%, and the mass concentration of EDC is 0.3-0.4 wt%.

[0022] Further, the volume ratio of the dopamine-modified double-bonded hyaluronic acid solution to the tyramine-modified hyaluronic acid solution in S3 is 1:0.5-0.5:1.

[0023] Further, the process of the enzymatic reaction is:

[0024] The hydrogel precursor liquid is divided into A liquid and B liquid, hydrogen peroxide is added to the A liquid, and horseradish peroxidase is added to the B liquid, then the A liquid and the B liquid are blended, and the hydrogel for promoting vascular intimal repair is prepared by using the enzymatic reaction.

[0025] Further, the concentration of hydrogen peroxide is 0.009%-0.019%, and the concentration of horseradish peroxidase is 10-30 U / mL.

[0026] Another object of the present application is to provide a hydrogel for promoting vascular intimal repair and having high adhesion, which is prepared by using the above method.

[0027] The present application has the following advantages:

[0028] 1. The present application provides a method for constructing a hydrogel, in which a gel precursor liquid containing hydrogen peroxide is coated on a surface, and then a gel precursor liquid containing an enzyme is coated on the surface to polymerize into a gel. The method is simple in process, does not need an external light curing device, and the constructed hydrogel has a structure similar to that of an extracellular matrix and has good biocompatibility.

[0029] 2. The hydrogel prepared by using the method of the present application can promote the adhesion, growth and migration of endothelial cells, and thus can promote the repair of vascular intima. Moreover, the hydrogel can inhibit the adhesion and aggregation and activation of platelets, and has excellent anticoagulant properties.

[0030] 3. The method of the present application is simple in process, does not need an additional light curing device, and the constructed hydrogel is similar to an extracellular matrix. The hydrogel can be coated into different shapes to adapt to different environments. Moreover, the hydrogel is rapidly gelled by using an enzymatic reaction, has good adhesion to tissues, has good biocompatibility, and is suitable for application in vascular repair after rotational atherectomy. BRIEF DESCRIPTION OF DRAWINGS

[0031] Figure 1 The Fourier transform infrared spectroscopy analysis diagram of dopamine-modified double-bonded hyaluronic acid and tyramine-modified hyaluronic acid prepared in Example 1;

[0032] Figure 2 The nuclear magnetic resonance hydrogen spectrum analysis diagram of dopamine-modified double-bonded hyaluronic acid and tyramine-modified hyaluronic acid prepared in Example 1;

[0033] Figure 3 The gelation and tissue adhesion test diagram of the gel precursor liquid prepared in the present application;

[0034] Figure 4 The adhesion and activation test of platelets on the hydrogel prepared in the present application;

[0035] Figure 5Endothelial cell adhesion detection is performed on the hydrogel prepared in the present application;

[0036] Figure 6 Endothelial cell growth detection is performed on the hydrogel prepared in the present application;

[0037] Figure 7 Endothelial cell migration detection is performed on the hydrogel prepared in the present application. DETAILED DESCRIPTION

[0038] The specific embodiments of the present application are described below to facilitate the understanding of the present application for those skilled in the art, but it should be clear that the present application is not limited to the scope of the specific embodiments, and for those skilled in the art, it is obvious that various changes are within the spirit and scope of the present application defined and determined by the appended claims, and all the inventions utilizing the concept of the present application are within the scope of protection.

[0039] Example 1

[0040] A high-adhesion enzyme-catalyzed hydrogel for promoting vascular intimal repair, the preparation method comprising the following steps:

[0041] S1, synthesis of dopamine-modified double-bonded hyaluronic acid

[0042] Methacrylic anhydride is added to the hyaluronic acid solution, the pH is adjusted to 8, the reaction is carried out for 3h, ethanol is added for precipitation, the precipitate is washed with alcohol, dissolved in deionized water, dialyzed and freeze-dried. After freeze-drying, dissolve in MES buffer to obtain a solution with a mass concentration of 1wt%, then add NHS, EDC and DOPA, remove oxygen, adjust the pH to 5.5, react for 4h, then adjust the pH of the system to be acidic, dialyze and freeze-dry;

[0043] The concentrations of the components in the above process are: hyaluronic acid 1wt%, methacrylic anhydride 4.5wt%, MES 100mM, NHS 0.288wt%, EDC 0.388wt%, and DOPA 0.471wt%.

[0044] S2, synthesis of tyramine-modified hyaluronic acid

[0045] After removing oxygen from the hyaluronic acid solution, add NHS, EDC and tyramine hydrochloride, adjust the pH to 4.5, react for 4h, dialyze and freeze-dry;

[0046] The concentrations of the components in the above process are: hyaluronic acid 1wt%, NHS 0.288wt%, EDC 0.388wt%, and tyramine hydrochloride 1.25mM;

[0047] S3, the dopamine modified double-bonded hyaluronic acid, tyramine modified hyaluronic acid and deionized water are mixed to prepare a hydrogel precursor solution with a mass concentration of 2wt% of dopamine modified double-bonded hyaluronic acid and tyramine modified hyaluronic acid; and the volume ratio of HMD solution to HT solution is 1:2;

[0048] S4, the hydrogel precursor solution is evenly divided into A and B, hydrogen peroxide is added to A, and horseradish peroxidase is added to B, wherein the concentration of hydrogen peroxide is 0.009%, and the concentration of horseradish peroxidase is 20U / mL.

[0049] S5, the A and B solutions are blended to prepare a high-adhesion enzyme hydrogel for promoting vascular intimal repair by using an enzymatic reaction.

[0050] Example 2

[0051] A high-adhesion enzyme hydrogel for promoting vascular intimal repair, the preparation method comprising the following steps:

[0052] S1, synthesis of dopamine modified double-bonded hyaluronic acid

[0053] Methyl acrylate is added to the hyaluronic acid solution, the pH is adjusted to 8, the reaction is carried out for 3h, ethanol is added for precipitation, the precipitate is washed with alcohol, dissolved in deionized water, dialyzed and freeze-dried; after freeze-drying, the solution is dissolved in MES buffer to obtain a solution with a mass concentration of 1.5wt%, then NHS, EDC and DOPA are added, oxygen is removed, the pH is adjusted to 5.5, the reaction is carried out for 4h, then the pH of the system is adjusted to be acidic, dialyzed and freeze-dried;

[0054] The concentrations of the components in the above process are as follows: hyaluronic acid 1.5wt%, methyl acrylate 5wt%, MES 120mM, NHS 0.288wt%, EDC 0.388wt%, and DOPA 0.5wt%.

[0055] S2, synthesis of tyramine modified hyaluronic acid

[0056] After the hyaluronic acid solution is deoxygenated, NHS, EDC and tyramine hydrochloride are added, the pH is adjusted to 4.5, the reaction is carried out for 4h, and then dialyzed and freeze-dried;

[0057] The concentrations of the components in the above process are as follows: hyaluronic acid 1wt%, NHS 0.288wt%, EDC 0.388wt%, and tyramine hydrochloride 1.5mM;

[0058] S3, the dopamine modified double-bonded hyaluronic acid, tyramine modified hyaluronic acid and deionized water are mixed to prepare a hydrogel precursor solution with a mass concentration of 2wt% of dopamine modified double-bonded hyaluronic acid and tyramine modified hyaluronic acid; and the volume ratio of HMD solution to HT solution is 1:2;

[0059] S4, the hydrogel precursor liquid is divided into A and B two parts, A liquid adds hydrogen peroxide, B liquid adds horseradish peroxidase, wherein the concentration of hydrogen peroxide is 0.009%, the concentration of horseradish peroxidase is 20U / mL.

[0060] S5, A and B liquid blending utilizes enzymatic reaction to prepare a high-adhesion enzymatic hydrogel for promoting vascular intima repair.

[0061] Example 3

[0062] A high-adhesion enzymatic hydrogel for promoting vascular intima repair, the preparation method comprising the following steps:

[0063] S1, synthesis of dopamine-modified double-bonded hyaluronic acid

[0064] Add methacrylic anhydride to the hyaluronic acid solution, adjust the pH to 8, react for 3h, add ethanol to precipitate, alcohol wash the precipitate, dissolve in deionized water, dialyze and freeze-dry; after freeze-drying, dissolve in MES buffer to obtain a solution with a mass concentration of 1wt%, then add NHS, EDC and DOPA, deoxygenate, adjust the pH to 5.5, react for 4h, then adjust the pH of the system to be acidic, dialyze and freeze-dry;

[0065] The concentrations of the components in the above process are: hyaluronic acid 1wt%, methacrylic anhydride 4.5wt%, MES 110mM, NHS 0.288wt%, EDC 0.388wt%, and DOPA 0.3wt%.

[0066] S2, synthesis of tyramine-modified hyaluronic acid

[0067] After deoxygenating the hyaluronic acid solution, add NHS, EDC and tyramine hydrochloride, adjust the pH to 4.5, react for 4h, dialyze and freeze-dry;

[0068] The concentrations of the components in the above process are: hyaluronic acid 1wt%, NHS 0.288wt%, EDC 0.388wt%, and tyramine hydrochloride 1.2mM;

[0069] S3, mix dopamine-modified double-bonded hyaluronic acid and tyramine-modified hyaluronic acid with deionized water to prepare a hydrogel precursor liquid with a mass concentration of 3wt% for each of dopamine-modified double-bonded hyaluronic acid and tyramine-modified hyaluronic acid; and the volume ratio of HMD solution to HT solution is 2:1;

[0070] S4, the hydrogel precursor liquid is divided into A and B two parts, A liquid adds hydrogen peroxide, B liquid adds horseradish peroxidase, wherein the concentration of hydrogen peroxide is 0.009%, the concentration of horseradish peroxidase is 20U / mL.

[0071] S5, A and B liquid blending uses enzymatic reaction to prepare a high adhesion enzymatic hydrogel for promoting vascular intimal repair.

[0072] Example 4

[0073] A high adhesion enzymatic hydrogel for promoting vascular intimal repair, the preparation method comprising the following steps:

[0074] S1, synthesis of dopamine-modified double-bonded hyaluronic acid

[0075] Add methacrylic anhydride to the hyaluronic acid solution, adjust the pH to 8, react for 2 hours, add ethanol to precipitate, alcohol wash the precipitate, dissolve in deionized water, dialyze and freeze-dry; after freeze-drying, dissolve in MES buffer to obtain a solution with a mass concentration of 1wt%, then add NHS, EDC and DOPA, remove oxygen, adjust the pH to 5.5, react for 4 hours, then adjust the pH of the system to be acidic, dialyze and freeze-dry;

[0076] The concentrations of the components in the above process are: hyaluronic acid 1wt%, methacrylic anhydride 5wt%, MES 100mM, NHS 0.28wt%, EDC 0.38wt%, and DOPA 0.46wt%;

[0077] S2, synthesis of tyramine-modified hyaluronic acid

[0078] After removing oxygen from the hyaluronic acid solution, add NHS, EDC and tyramine hydrochloride, adjust the pH to 5, react for 8 hours, dialyze and freeze-dry;

[0079] The concentrations of the components in the above process are: hyaluronic acid 1wt%, NHS 0.28wt%, EDC 0.38wt%, and tyramine hydrochloride 1.2mM.

[0080] S3, mix dopamine-modified double-bonded hyaluronic acid, tyramine-modified hyaluronic acid and deionized water to prepare a hydrogel precursor solution with a mass concentration of 3wt% for both dopamine-modified double-bonded hyaluronic acid and tyramine-modified hyaluronic acid; and the volume ratio of HMD solution to HT solution is 1:1;

[0081] S4, divide the hydrogel precursor solution into A and B two parts, add hydrogen peroxide to A, and add horseradish peroxidase to B, wherein the concentration of hydrogen peroxide is 0.019%, and the concentration of horseradish peroxidase is 10U / mL;

[0082] S5, A and B liquid blending uses enzymatic reaction to prepare a high adhesion enzymatic hydrogel for promoting vascular intimal repair.

[0083] Example 5

[0084] A high adhesion enzymatic hydrogel for promoting vascular intimal repair, the preparation method comprising the following steps:

[0085] S1, synthesis of dopamine-modified double-bonded hyaluronic acid

[0086] Add methacrylic anhydride to the hyaluronic acid solution, adjust the pH to 8, react for 3 h, add ethanol to precipitate, wash the precipitate with alcohol, dissolve in deionized water, freeze-dry after dialysis; dissolve in MES buffer after freeze-drying to obtain a solution with a mass concentration of 1 wt%, then add NHS, EDC, DOPA, remove oxygen, adjust the pH to 5.5, react for 4 h, then adjust the pH of the system to be acidic, freeze-dry after dialysis;

[0087] The concentrations of the components in the above process are: hyaluronic acid 1 wt%, methacrylic anhydride 5 wt%, MES 100 mM, NHS 0.29 wt%, EDC 0.39 wt%, and DOPA 0.48 wt%;

[0088] S2, synthesis of tyramine-modified hyaluronic acid

[0089] After removing oxygen from the hyaluronic acid solution, add NHS, EDC, and tyramine hydrochloride, adjust the pH to 5, react for 4 h, and freeze-dry after dialysis;

[0090] The concentrations of the components in the above process are: hyaluronic acid 1 wt%, NHS 0.29 wt%, EDC 0.39 wt%, and tyramine hydrochloride 1.3 mM.

[0091] S3, mix dopamine-modified double-bonded hyaluronic acid, tyramine-modified hyaluronic acid, and deionized water to prepare a hydrogel precursor solution with a mass concentration of 2 wt% for each of dopamine-modified double-bonded hyaluronic acid and tyramine-modified hyaluronic acid; and the volume ratio of HMD solution to HT solution is 1:1;

[0092] S4, divide the hydrogel precursor solution into two parts A and B, add hydrogen peroxide to part A and horseradish peroxidase to part B, wherein the concentration of hydrogen peroxide is 0.019% and the concentration of horseradish peroxidase is 30 U / mL;

[0093] S5, blend parts A and B to prepare a high-adhesion enzyme hydrogel for promoting vascular intimal repair by enzyme reaction.

[0094] Test example

[0095] Take the preparation method in Example 1 as an example to test the product, as follows:

[0096] 1. Fourier transform infrared spectroscopy analysis

[0097] Grind and press the raw material hyaluronic acid, dopamine-modified double-bonded hyaluronic acid, and tyramine-modified hyaluronic acid prepared in Example 1 of the present application in potassium bromide (KBr). Through FT-IR at 4000-500 cm -1Infrared absorption peaks were obtained at wavenumber.

[0098] like Figure 1 As shown, compared to hyaluronic acid, it can be used at 1262cm. -1 The observation of CO stretching vibrations in phenol indicates that dopamine and tyramine were successfully grafted onto the molecular chain of hyaluronic acid.

[0099] 2. Proton NMR spectroscopy analysis

[0100] The raw material hyaluronic acid, the dopamine-modified double-bonded hyaluronic acid and the tyramine-modified hyaluronic acid prepared in Example 1 of this invention were dissolved in deuterated water and subjected to nuclear magnetic resonance hydrogen spectroscopy analysis to obtain the nuclear magnetic resonance hydrogen spectra characterizing the structure.

[0101] like Figure 2 As shown, the analysis results of the MestReNova software indicate that the NMR peak at the chemical shift δ = 2.95 ppm is a characteristic peak of the two methylene groups of dopamine and tyramine attached to the benzene ring, indicating that dopamine and tyramine were successfully grafted onto the molecular chain of hyaluronic acid.

[0102] 3. Glue formation and tissue adhesion test

[0103] The hydrogel prepared in Example 1 was applied to the inner wall of the rabbit thoracic aorta, where it gelled and adhered firmly. Subsequent torsion, stretching, and water rinsing of the gel coating showed that it remained firmly adhered to the tissue without breaking or detaching. Figure 3 As shown, the coating prepared from the gel precursor solution of the present invention exhibits high adhesion to wet tissues.

[0104] 4. Platelet adhesion and activation

[0105] Platelet adhesion and activation tests were performed on the hydrogel coatings prepared in Examples 1-3 of this invention. The number and morphology of platelets were observed by scanning electron microscopy. The specific test methods are as follows:

[0106] After co-incubating the rich-slab slurry with the hydrogels prepared in Examples 1-3, the hydrogels were fixed with 2.5% glutaraldehyde solution for 24 hours, followed by gradient dehydration using 30-100% ethanol aqueous solutions. After drying, the hydrogels were observed under a scanning electron microscope. The results are shown in the figure. Figure 4 In the figure, HMDHT0.5 is the hydrogel prepared in Example 1, HMDHT1 is the hydrogel prepared in Example 2, and HMDHT2 is the hydrogel prepared in Example 3.

[0107] like Figure 4 As shown, compared with the 316L stainless steel group, the amount of platelet adhesion is significantly reduced and the platelet activation is less, indicating that the hydrogel coating prepared by the present invention has excellent anticoagulant effect.

[0108] 5. Endothelial cell adhesion, growth and migration

[0109] Endothelial cell adhesion, growth and migration tests were performed on the gel coatings prepared in Examples 1-3 of the present application, and the results are shown in Figures 5 to 7 , and the specific process is as follows:

[0110] (1) Endothelial cell adhesion test:

[0111] The prepared hydrogel product was sterilized and placed in a 24-well plate, 500 μl of tracer-stained endothelial cell suspension (cell density of about 25,000 cells per well) was added, and after 4 h of incubation in an incubator, the surface was gently rinsed and photographed using a confocal microscope.

[0112] (2) Cck-8 test:

[0113] Endothelial cells were seeded at 0.5 million cells per well in a 48-well plate, and after adhering, the culture medium was replaced with a hydrogel product culture medium extract. After 1 day and 3 days, the culture medium was aspirated, 200 μl of cck-8 solution was added per well, incubated for 2 h, and then 100 μl of the culture medium was aspirated into a 96-well plate to measure the absorbance value of the solution using a microplate reader.

[0114] (3) Cell migration test:

[0115] An L-shaped stainless steel sheet was sterilized after being coated with a hydrogel product on one side, and the side of the stainless steel sheet not coated with the hydrogel product was placed at the bottom of the well plate, and tracer-stained endothelial cells (25,000 per well) were seeded. After the cells adhered, the stainless steel sheet was moved to another 24-well plate so that the hydrogel side was at the bottom of the well plate, at which time the endothelial cells on the vertical surface would begin to migrate. After 24 hours, the cell migration image on the hydrogel side was photographed using a confocal microscope.

[0116] As shown in Figure 5 , it can be seen that the endothelial cell adhesion amount of the hydrogel coating group prepared by the present application is more than that of the control group. The Cck-8 results are shown in Figure 6 , indicating that the hydrogel coating group prepared by the present application can promote the growth of endothelial cells. The cell migration experiment results are shown in Figure 7 , from which it can be seen that the hydrogel coating group prepared by the present application can more promote the migration of endothelial cells.

[0117] Finally, it should be noted that the above specific embodiments are only used to illustrate the technical solutions of the present application and not to limit it. Although the present application has been described in detail with reference to the examples, those skilled in the art should understand that the technical solutions of the present application can be modified or replaced equivalently without departing from the spirit and scope of the technical solutions of the present application, which should be covered in the scope of the claims of the present application.

Claims

1. A method for preparing a highly adhesive enzyme-catalyzed hydrogel that promotes vascular intima repair, characterized in that, Includes the following steps: S1, Dopamine-modified double-bonded hyaluronic acid Hyaluronic acid was modified by double bonding with methacrylic anhydride and then crosslinked with dopamine using NHS / EDC as the crosslinking agent to prepare dopamine-modified double-bonded hyaluronic acid. S2, Tyrosine-modified hyaluronic acid After deoxygenating the hyaluronic acid solution, crosslinking agent NHS / EDC and tyramine hydrochloride are added, and the mixture is reacted in an acidic environment for 4-8 hours to obtain tyramine-modified hyaluronic acid. S3. Mix the dopamine-modified double-bonded hyaluronic acid solution with the tyramine-modified hyaluronic acid solution to obtain a hydrogel precursor solution with a mass concentration of 2-3 wt% for both dopamine-modified double-bonded hyaluronic acid and tyramine-modified hyaluronic acid. S4. A highly adhesive enzymatic hydrogel promoting vascular intimal repair is then prepared using an enzymatic reaction; the enzymatic reaction process is as follows: The hydrogel precursor solution is divided into solution A and solution B. Hydrogen peroxide is added to solution A and horseradish peroxidase is added to solution B. Then, solutions A and B are mixed together to obtain a hydrogel that promotes vascular endothelial repair through enzymatic reaction.

2. The preparation method according to claim 1, characterized in that, The process of double-bonding hyaluronic acid using methacrylic anhydride is as follows: Methacrylic anhydride was added to a hyaluronic acid solution to adjust the pH to 8-9. The reaction was allowed to proceed for 2-3 hours. Ethanol was then added to precipitate the precipitate. After washing the precipitate with alcohol, it was dissolved in deionized water and then lyophilized by dialyzing. The hyaluronic acid has a mass concentration of 1-1.5 wt%, and the methacrylic anhydride has a mass concentration of 4.5-5 wt%.

3. The preparation method according to claim 1 or 2, characterized in that, The process of dopamine-modified double-bond hyaluronic acid is as follows: Double-bonded modified hyaluronic acid was dissolved in MES buffer to obtain a solution with a mass concentration of 1~1.5wt%. Then, cross-linking agent NHS / EDC and dopamine were added. After deoxygenation, the pH was adjusted to 5~6. After reacting for 4~8 hours, the pH was adjusted to acidic, and the solution was lyophilized by dialyzing. The concentration of MES buffer is 100-120 mM, and the mass concentration of dopamine is 0.3-0.5 wt%.

4. The preparation method according to claim 3, characterized in that, The mass concentration of NHS is 0.2~0.3wt%, and the mass concentration of EDC is 0.3~0.4wt%.

5. The preparation method according to claim 1, characterized in that, The mass concentration of hyaluronic acid in S2 is 0.5~1.5wt%, and the concentration of tyramine hydrochloride is 1.2~1.5mM.

6. The preparation method according to claim 1, characterized in that, The mass concentration of NHS in S2 is 0.2~0.3wt%, and the mass concentration of EDC is 0.3~0.4wt%.

7. The preparation method according to claim 1, characterized in that, The volume ratio of dopamine-modified double-bonded hyaluronic acid solution to tyramine-modified hyaluronic acid solution in S3 is 1:0.5~0.5:

1.

8. The preparation method according to claim 1, characterized in that, The hydrogen peroxide concentration was 0.009%-0.019%, and the horseradish peroxidase concentration was 10-30 U / mL.

9. A highly adhesive enzyme-catalyzed hydrogel that promotes vascular intima repair, characterized in that, It is prepared by the method described in any one of claims 1 to 8.

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