CN116535699A

CN116535699A - Bionic immune regulation double-network hydrogel and preparation method and application thereof

Info

Publication number: CN116535699A
Application number: CN202310500392.7A
Authority: CN
Inventors: 孙明明; 于艳
Original assignee: China Institute Of Rehabilitation Sciences Research Center For Disability Prevention And Control Of China Disabled Persons' Federation
Current assignee: China Institute Of Rehabilitation Sciences Research Center For Disability Prevention And Control Of China Disabled Persons' Federation
Priority date: 2023-05-06
Filing date: 2023-05-06
Publication date: 2023-08-04

Abstract

The invention discloses a bionic immune regulation double-network hydrogel and a preparation method and application thereof, wherein the preparation method comprises the following steps: dissolving GelMA-SPD and HAMA-NB, adding a photo-crosslinking agent, and crosslinking under illumination to obtain double-network hydrogel; the GelMA-SPD is methacrylic acid gelatin-spermidine; the HAMA-NB is methacrylic acid N- (2-aminoethyl) -4- [4- (hydroxymethyl) -2-methoxy-5-nitrophenoxy ] -butyramide. The hydrogel prepared by the invention has good biocompatibility, simple preparation process and degradability. The hydrogel can be applied to tissue repair materials or tissue engineering scaffolds, is hopeful to solve the problem of immune rejection in the process of tissue and organ transplantation, is beneficial to reducing inflammatory reaction and promoting rapid healing of tissue wounds.

Description

Bionic immune regulation double-network hydrogel and preparation method and application thereof

Technical Field

The invention relates to the technical field of biomedical materials and related medical instrument research and development, in particular to bionic immune regulation hydrogel and a preparation method and application thereof.

Background

About billions of people face different forms of tissue and organ injury each year worldwide, and as the aging rate of the population increases year by year, more and more patients with chronic degenerative diseases are suffering. Spinal cord tissue and brain tissue play an important role in nerve signal transduction, and play a key role in normal physiological functions of the body. Spinal Cord Injury (SCI) is a serious traumatic disorder of the central nervous system, which causes temporary or permanent loss of motor function in patients, severely degrading the quality of life of the patients. SCI patients are not only faced with the disorders of daily life caused by neurological deficit, but also are burdened with expensive treatment costs, bearing a great mental burden. Spinal cord injury is one of the most disabling and damaging neurological diseases, and because of poor central nervous system plasticity, the ability of neurons to regenerate is limited, and the regenerative repair after spinal cord injury is very weak.

At present, the clinical treatment of spinal cord injury is limited to operation decompression, hormone impact therapy, neuroprotection treatment and the like, the clinical curative effect is very low, the patient maintains the remained nerve function mainly through later rehabilitation training, and no effective clinical treatment measures are available to better recover the nerve function lost after spinal cord injury. Spinal cord injury repair and functional reconstruction have become significant medical problems worldwide, and are a significant challenge facing the global health field.

The medical hydrogel is used as a novel tissue engineering scaffold, has a three-dimensional network structure, contains a large amount of moisture, and can provide a suitable microenvironment for cell proliferation; at the same time, the local inflammatory reaction is limited, the apoptosis is inhibited, and the repair and regeneration of tissues are promoted.

After the tissue is damaged, the local microenvironment is obviously changed into a severe inflammatory microenvironment, so that the loss of extracellular matrix is further aggravated, and the damage is aggravated. In addition, local inflammation promotes the formation of glial scars, impeding axon regeneration. Therefore, biomaterial scaffolds that can modulate local microenvironments hold promise for repair and regeneration of spinal cord injury. At present, hyaluronic acid hydrogel, PLGA hydrogel, HEMA hydrogel, nanofiber, self-assembled peptide hydrogel and the like can structurally provide support for cell migration and axon regeneration. However, most hydrogels are also very different from the autologous environment, cannot well meet the condition of nerve regeneration, and have obvious rejection phenomenon in vivo, so that more damage is caused to tissues. Dynamic hydrogels with cellular adaptability can provide a better microenvironment for proliferation, differentiation, and recruitment of immune cells. Therefore, the development of the double-network hydrogel with cell adaptability and rejection resistance not only can improve the mechanical property, but also can provide a good microenvironment for proliferation and differentiation of cells, and is beneficial to tissue repair and regeneration.

Therefore, how to provide a dual-network hydrogel with high safety and immune regulation is a problem to be solved by those skilled in the art.

Disclosure of Invention

In view of the above, the invention provides a preparation method of bionic immune-regulating double-network hydrogel. The hydrogel prepared from hyaluronic acid, gelatin and spermidine has good biocompatibility and immune regulation effect; the hydrogel prepared by the method has simple preparation process, is degradable and has good application prospect.

In order to achieve the above purpose, the present invention adopts the following technical scheme:

a preparation method of bionic immune regulation dual-network hydrogel comprises dissolving GelMA-SPD and HAMA-NB, adding photocrosslinker, and crosslinking under illumination to obtain dual-network hydrogel; the GelMA-SPD is methacrylic acid gelatin-spermidine; the HAMA-NB is methacrylic acid hyaluronic acid-4- (4- (hydroxymethyl) -2-methoxy-5-nitrophenoxy) butyramide.

The hydrogel is based on a double-network structure design of ultraviolet light curing, spermidine is introduced into the double-network hydrogel through a dynamic covalent bond, and can slowly release the spermidine along with the degradation of the hydrogel, so that the biological effect is exerted for a long time. Under ultraviolet light, NB generates aldehyde group, then reacts with amino group, and a primary network is formed through Schiff base reaction; the two-stage network is formed by free radical polymerization reaction of carbon-carbon double bonds, and the two-stage network hydrogel is formed by combining the two-stage network hydrogel.

The Schiff base reaction and the free radical polymerization reaction have the advantages of high speed and mild reaction conditions, and the preparation method of the hydrogel is simple and feasible, high-efficiency and convenient, is easy to realize large-scale production, and has wide application prospect in the biomedical hydrogel field. Gelatin, hyaluronic acid and spermidine are all biological endogenous substances, and degradation products are nontoxic and easy to be discharged out of the body. The double-network hydrogel prepared by the invention can regulate and control immune response, reduce inflammation and relieve rejection response, and can be used for tissue repair scaffolds to promote tissue repair and regeneration.

Preferably, the preparation method of GelMA comprises the following steps: dissolving gelatin in phosphate buffer solution, regulating pH to 7.4-11.0, heating to 50-70 ℃ for full dissolution, then adding methacrylic anhydride, stirring for reaction for 6-24 h, dialyzing for 48-96h, and freeze-drying for 48-96h to obtain methacrylic gelatin GelMA; the concentration of the gelatin in the phosphate buffer solution is 1-10wt%, and the mol ratio of the gelatin to methacrylic anhydride is 1:

(2～5)：(2～5)：(1～10)。

preferably, the preparation method of the GelMA-SPD comprises the following steps: dissolving GelMA in distilled water with the concentration of 1-10wt%, and adding 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide into the distilled water according to the molar ratio of EDC, N-hydroxysuccinimide NHS, spermidine SPD, EDC, NHS, gelMA and SPD of 1: (1-10), stirring and reacting for 6-72 h, dialyzing for 48-96h, and freeze-drying for 48-96h to obtain GelMA-SPD.

Preferably, the preparation method of HAMA comprises the following steps: dissolving hyaluronic acid HA in distilled water to a concentration of 0.1-5wt%, adding methacrylic anhydride, stirring and reacting for 12-48 h, wherein the molar ratio of HA to methacrylic anhydride is 1: (1-10), dialyzing for 48-96h, and freeze-drying for 48-96h to obtain the methacrylated hyaluronic acid HAMA.

Preferably, the HAMA-NB is prepared by the following steps: dissolving HAMA in distilled water to a concentration of 0.1-5wt%, and then adding 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride EDC, N-hydroxysuccinimide NHS and 4- (4- (hydroxymethyl) -2-methoxy-5-nitrophenoxy) butyramide NB, wherein the molar ratio of the HAMA, EDC, NHS and NB is 1: (1-5): (1-5): (1-5), reacting for 6-24 h, dialyzing for 48-96h, and freeze-drying for 48-96h to obtain HAMA-NB.

Preferably, the GelMA-SPD and the HAMA-NB are dissolved in distilled water, the concentration is 1 to 20 weight percent, and a photo-crosslinking agent is added, wherein the molar ratio of the GelMA-SPD, the HAMA-NB and the photo-crosslinking agent is (1 to 10):

(0.1-10): (0.1-1), and crosslinking under illumination to obtain the double-network hydrogel.

Preferably, the dialysis parameters in the preparation process of GelMA-SPD and HAMA-NB are dialysis bags of 3kDa to 20kDa, and then the GelMA-SPD and the HAMA-NB are frozen in an environment of minus 10 ℃ to minus 80 ℃ and then are dried in a freeze dryer.

Preferably, the photocrosslinker is selected from any one of I2959, LAP, TPO, MBP, benzophenone, thiopropylthioxanthone, benzoin and fluorinated diphenyl titanocene, and the illumination condition is as follows: wavelength 250-800 nm, power 2-30W, photo-crosslinking time 5-30 s.

The beneficial effect of adopting above-mentioned technical scheme: the reaction is carried out at low temperature, the reaction condition is mild, and the damage to gelatin is reduced to the greatest extent.

The invention also provides application of the bionic immune regulation dual-network hydrogel in the aspects of tissue repair materials or tissue engineering scaffolds.

The invention has the beneficial effects that: according to the invention, two polymer materials are dissolved in distilled water, and the double-network hydrogel is prepared through photo-crosslinking, and the spermidine is slowly released through degradation, so that the local concentration is reduced, the toxic and side effects are lightened, and the double-network hydrogel has excellent bioactivity, biocompatibility and biodegradability. Meanwhile, the chemical reaction related by the invention has the advantages of high speed, mild reaction conditions and the like, so that the hydrogel preparation method is simple and feasible, high-efficiency and convenient, can realize batch production, and has wide application prospect in the biomedical hydrogel field. The hydrogel can be applied to tissue repair materials or tissue engineering scaffolds, is hopeful to solve the problem of immune rejection in the process of tissue and organ transplantation, is beneficial to reducing inflammatory reaction and promoting rapid healing of tissue wounds.

Description of the drawings:

FIG. 1 is a synthetic route for GelMA-SPD;

FIG. 2 is a synthetic route for HAMA-NB;

FIG. 3 is a preparation route of the bionic immune-modulating dual-network hydrogel;

FIG. 4 is a nuclear magnetic pattern of GelMA-SPD and HAMA-NB;

FIG. 5 is a bionic immune-modulating dual-network hydrogel physical diagram;

FIG. 6 is an SEM image of a biomimetic immunomodulatory dual network hydrogel;

FIG. 7 is a statistical graph of the cellular activity of endothelial cells (HUVECs) of the present invention in a biomimetic immunomodulatory dual-network hydrogel leach;

FIG. 8 is a graph showing the mobility of endothelial cells (HUVECs) of the present invention on the surface of biomimetic immunomodulatory dual-network hydrogels;

FIG. 9 shows the cell formation rate of endothelial cells (HUVECs) of the present invention on the surface of biomimetic immunomodulatory dual-network hydrogels;

FIG. 10 shows the wound repair of damaged skin of a mouse according to the invention at different time points on the surface of a biomimetic immunoregulatory dual-network hydrogel;

FIG. 11 shows the biocompatibility of the biomimetic immunomodulatory dual-network hydrogel of the present invention after implantation into the skin.

The specific embodiment is as follows:

the technical scheme of the invention is further specifically described below through examples and with reference to the accompanying drawings.

Example 1:

a preparation method of bionic immune-regulating double-network hydrogel comprises the following steps:

s1: dissolving 1moL of gelatin and 1moL of methacrylic anhydride in 100mL of phosphate solution, adjusting the pH value to 7.4, heating to 50 ℃, reacting for 5 hours under the dark condition, adding distilled water to terminate the reaction, dialyzing for 72 hours, and freeze-drying to obtain methacrylic gelatin (GelMA);

s2: dissolving 1molgelMA in 100mL of distilled water, adding 2 mols of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), 2 moLN-hydroxysuccinimide (NHS) and 1 mols of Spermidine (SPD), reacting for 24 hours at room temperature, dialyzing for 72 hours, and freeze-drying for 48 hours to obtain GelMA-SPD;

s3: dissolving 1moL of hyaluronic acid and 1moL of methacrylic anhydride in 100mL of distilled water, reacting for 24 hours at room temperature, dialyzing for 48 hours, and freeze-drying for 48 hours to obtain methacrylic hyaluronic acid (HAMA);

s4: 1molH AMA, 1molN- (2-amino ethyl) -4- (4-hydroxymethyl) -2-methoxy-5-Nitrosoethoxy) Butyramide (NB), 2molN 1-ethyl-3- (3-dimethyl amino propyl) carbodiimide hydrochloride (EDC) and 2 molN-hydroxysuccinimide (NHS) are dissolved in 100mL of distilled water, reacted for 24 hours at room temperature, dialyzed for 48 hours and freeze-dried to obtain HAMA-NB;

s5: mixing and dissolving 0.1molgelMA-SPD, 0.01 molgelMA-NB and 0.001 molleap in 10mL distilled water, stirring and mixing uniformly at room temperature, standing for degassing, and crosslinking for 5s under 405nm blue light to obtain hydrogel.

In the steps S1-S4, dialysis parameters are that a dialysis bag with a molecular weight of 3000 Da-20 kDa is adopted for dialysis for 72 hours, and then the dialysis is carried out in an environment with the temperature of minus 10 ℃ for freezing.

Example 2:

s1: dissolving 1moL of gelatin and 2moL of methacrylic anhydride in 100mL of phosphate solution, adjusting the pH value to 7.4, heating to 60 ℃, reacting for 8 hours under the dark condition, adding distilled water to terminate the reaction, dialyzing for 96 hours, and freeze-drying to obtain methacrylic gelatin (GelMA);

s2: dissolving 1molgelMA in 100mL of distilled water, adding 3 mols of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), 3 moLN-hydroxysuccinimide (NHS) and 2 mols of Spermidine (SPD), reacting for 48h at room temperature, dialyzing for 96h, and freeze-drying for 72h to obtain GelMA-SPD;

s3: dissolving 1moL of hyaluronic acid and 2moL of methacrylic anhydride in 100mL of distilled water, reacting for 36h at room temperature, dialyzing for 96h, and freeze-drying for 72h to obtain methacrylic hyaluronic acid (HAMA);

s4: 1molH AMA, 3molN- (2-amino ethyl) -4- (4-hydroxymethyl) -2-methoxy-5-Nitrosoethoxy) Butyramide (NB), 3molN 1-ethyl-3- (3-dimethyl amino propyl) carbodiimide hydrochloride (EDC) and 3 molN-hydroxysuccinimide (NHS) are dissolved in 100mL of distilled water, reacted for 72 hours at room temperature, dialyzed for 72 hours and freeze-dried to obtain HAMA-NB;

s5: mixing and dissolving 0.2molgelMA-SPD, 0.1 molleama-NB and 0.01 molleap in 20mL distilled water, stirring and mixing uniformly at room temperature, standing for degassing, and crosslinking for 10s under 405nm blue light to obtain hydrogel.

In the steps S1-S4, dialysis parameters are that dialysis is carried out for 72 hours by using a dialysis bag with the molecular weight of 3000 Da-20 kDa, and then the dialysis is carried out in an environment with the temperature of minus 40 ℃.

Example 3:

s1: dissolving 1moL of gelatin and 2moL of methacrylic anhydride in 100mL of phosphate solution, adjusting the pH value to 11, heating to 70 ℃, reacting for 12 hours under the condition of avoiding light, adding distilled water to terminate the reaction, dialyzing for 120 hours, and freeze-drying to obtain methacrylic gelatin (GelMA);

s2: dissolving 1molgelMA in 100mL of distilled water, adding 5moL of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), 5 moLN-hydroxysuccinimide (NHS) and 10moL of Spermidine (SPD), reacting at room temperature for 72h, dialyzing for 120h, and freeze-drying for 96h to obtain GelMA-SPD;

s3: 1moL of hyaluronic acid and 5moL of methacrylic anhydride are dissolved in 200mL of phosphate solution, the pH is adjusted to 11, the mixture is reacted for 48 hours at room temperature, dialyzed for 120 hours and freeze-dried for 120 hours, and the methacrylic hyaluronic acid (HAMA) is obtained;

s4: 1molH AMA, 5molN- (2-amino ethyl) -4- (4-hydroxymethyl) -2-methoxy-5-Nitrosoethoxy) Butyramide (NB), 3molN 1-ethyl-3- (3-dimethyl amino propyl) carbodiimide hydrochloride (EDC) and 3 molN-hydroxysuccinimide (NHS) are dissolved in 200mL of distilled water, reacted for 72h at room temperature, dialyzed for 120h and freeze-dried for 96h to obtain HAMA-NB;

s5: 0.1moLGelMA-SPD (10%), 0.11 moLGelMA-NB and 0.01 molleap are mixed and dissolved in 10mL distilled water, stirred and mixed uniformly at room temperature, stood still for deaeration, and crosslinked for 60s under 405nm blue light to obtain hydrogel.

In the steps S1-S4, dialysis parameters are that a dialysis bag with a molecular weight of 3000 Da-20 kDa is adopted for dialysis for 120 hours, and then the dialysis is carried out in an environment with the temperature of minus 80 ℃ for freezing.

Test example:

1. ¹ HNMR analysis

The chemical structures of the GelMA-SPD and the HAMA-NB prepared in the invention are proved to be correct by using a nuclear magnetic resonance spectrometer Bruker400 to test the GelMA-SPD and the HAMA-NB prepared in the embodiment 1, the nuclear magnetic resonance spectrum results are shown in figure 4, and the accuracy of the chemical structures of the GelMA-SPD and the HAMA-NB is proved by spectrum analysis.

2. Topography analysis

Fig. 5 and 6 are a physical and SEM image of the dual-network hydrogel prepared in example 1, respectively. As shown in fig. 6, the double-network hydrogel has a three-dimensional network structure inside.

3. Biocompatibility testing

1. In vitro cytotoxicity evaluation of hydrogels

The killing effect of hydrogels on endothelial cells (HUVECs) was examined using the CCK-8 method. The dual-network hydrogel prepared in example 1 (1:1, 10%) was co-cultured with cells, incubated for 2h, and then acted upon with CCK-8 solution (10. Mu.L, 5 mg/mL) for 4h. Absorbance at 450nm was measured and compared with untreated cells to calculate their cytotoxicity. As shown in FIG. 7, the cell viability was above 90%, demonstrating that the double-network hydrogel was not significantly cytotoxic to endothelial cells (HUVECs).

Reference GB/T16886.5-2017, section 5 of medical device biological evaluation: in vitro cytotoxicity experiments, the cell survival rate is not less than 70% and the cytotoxicity is classified as class I, which indicates that the hydrogel has no cytotoxicity.

2. Evaluation of hydrogel-promoted cell migration

The double-network hydrogel prepared in the example 1 is spread on a culture dish, then cells are inoculated on the culture dish, and the culture is carried out for 24 hours; then, the cell scratch is lightly manufactured by a straw, the cell scratch is put into an incubator for culture, and the migration condition of the cell is observed by an optical microscope, see figure 8, and the migration rate of the experimental group is 2 times that of the control group, which shows that the hydrogel can promote the migration of the cell and is beneficial to wound healing.

3. Hydrogel-promoted cell tube formation evaluation

The double-network hydrogel prepared in example 1 is coated on a cell culture plate, then cells are inoculated on the culture plate, the culture is continued in an incubator, and then the tube forming condition of the cells is observed under an optical microscope, as shown in fig. 9, the hydrogel can promote the tube forming of the cells, and has no obvious difference compared with matrigel, so that the double-network hydrogel has better effect on the tube forming of the cells.

4. In vivo skin injury repair test

SD rats (8 weeks, n=5) were subjected to general anesthesia and fixed on an operating table to create a full-thickness skin injury model. The wound repair cases at different time points were observed in a control group and an experimental group (double-network hydrogel prepared in example 1), as shown in fig. 10. The wound of the rats of the experimental group recovered well after the hydrogel treatment prepared in example 1. The hydrogel can promote the repair of skin wounds. The hydrogel has good capability of promoting tissue repair.

5. In vivo rejection response test

The double network hydrogel prepared in example 1 was implanted subcutaneously, had good biocompatibility in vivo, and had no significant fibrocyst formation, while the control group had significant fibrocyst formation.

In conclusion, the prepared hydrogel has better biocompatibility, promotes cell migration and tube formation, and promotes tissue repair and regeneration, and is a better biological scaffold.

The previous description of the embodiments is provided to facilitate a person of ordinary skill in the art in order to make and use the present invention. It will be apparent to those skilled in the art that various modifications can be readily made to these embodiments and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the embodiments described herein, and those skilled in the art, based on the present disclosure, should make improvements and modifications within the scope of the present invention.

Claims

1. A preparation method of bionic immune regulation double-network hydrogel is characterized in that GelMA-SPD and HAMA-NB are dissolved, a photo-crosslinking agent is added, and crosslinking is carried out under illumination, so that double-network hydrogel is obtained; the GelMA-SPD is methacrylic acid gelatin-spermidine; the HAMA-NB is methacrylic acid N- (2-aminoethyl) -4- [4- (hydroxymethyl) -2-methoxy-5-nitrophenoxy ] -butyramide.

2. The preparation method of the bionic immune-regulating double-network hydrogel according to claim 1, wherein the preparation method of GelMA is as follows: dissolving gelatin in phosphate buffer solution, regulating pH to 7.4-11.0, heating to 50-70 ℃ for full dissolution, then adding methacrylic anhydride, stirring for reaction for 6-24 h, dialyzing for 48-96h, and freeze-drying for 48-96h to obtain methacrylic gelatin GelMA; the concentration of the gelatin in the phosphate buffer solution is 1-10wt%, and the mol ratio of the gelatin to methacrylic anhydride is 1: (1-10).

3. The preparation method of the bionic immune-regulating double-network hydrogel according to claim 2, wherein the preparation method of the GelMA-SPD is as follows: dissolving GelMA in distilled water with the concentration of 1-10wt%, and adding 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide into the distilled water according to the molar ratio of EDC, N-hydroxysuccinimide NHS, spermidine SPD, EDC, NHS, gelMA and SPD of 1: (2-5): (2-5): (1-10), stirring and reacting for 6-72 h, dialyzing for 48-96h, and freeze-drying for 48-96h to obtain GelMA-SPD.

4. The method for preparing the bionic immune-regulating double-network hydrogel according to claim 1, wherein the preparation method of HAMA is as follows: dissolving hyaluronic acid HA in distilled water to a concentration of 0.1-5wt%, adding methacrylic anhydride, stirring and reacting for 12-48 h, wherein the molar ratio of HA to methacrylic anhydride is 1: (1-10), dialyzing for 48-96h, and freeze-drying for 48-96h to obtain the methacrylated hyaluronic acid HAMA.

5. The method for preparing the bionic immune-regulating double-network hydrogel according to claim 4, wherein the preparation method of HAMA-NB is as follows: HAMA was dissolved in distilled water at a concentration of 0.1wt% to 5wt% and then 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride EDC, N-hydroxysuccinimide NHS, N- (2-aminoethyl) -4- [4- (hydroxymethyl) -2-methoxy-5-nitrophenoxy ] -butyramide NB were added at a molar ratio of HAMA, EDC, NHS to NB of 1: (1-5): (1-5): (1-5), reacting for 6-24 h, dialyzing for 48-96h, and freeze-drying for 48-96h to obtain HAMA-NB.

6. The method for preparing the bionic immune-regulating dual-network hydrogel according to any one of claims 1 to 5, wherein GelMA-SPD and HAMA-NB are dissolved in distilled water, the concentration is 1 to 20 weight percent, a photo-crosslinking agent is added, and the molar ratio of GelMA-SPD, HAMA-NB and the photo-crosslinking agent is (1 to 10): (0.1-10): (0.1-1), and crosslinking under illumination to obtain the double-network hydrogel.

7. The method for preparing the bionic immune-regulating double-network hydrogel according to claims 1-5, wherein dialysis parameters in the preparation process of GelMA-SPD and HAMA-NB are dialysis bags of 3 kDa-20 kDa, and the dialysis bags are frozen in an environment of-10 ℃ to-80 ℃ for 72-120 hours and then dried in a freeze dryer.

8. The preparation method of the bionic immune-regulating dual-network hydrogel according to claim 1, wherein the photocrosslinker is selected from any one of I2959, LAP, TPO, MBP, benzophenone, thiopropylthioxanthone, benzoin and fluorinated diphenyl titanocene, and the illumination condition is as follows: wavelength 250-800 nm, power 2-30W, photo-crosslinking time 5-30 s.

9. A biomimetic immune-controlled dual-network hydrogel prepared by the method according to any one of claims 1-8.

10. Use of the biomimetic immune-modulating double-network hydrogel according to claim 9 in tissue repair materials or tissue engineering scaffolds.