CN117959238A

CN117959238A - Double-network interpenetrating hydrogel microneedle based on silk fibroin and acylated hyaluronic acid and preparation method thereof

Info

Publication number: CN117959238A
Application number: CN202410018398.5A
Authority: CN
Inventors: 黄颖; 华佳慧; 张强; 闫书芹; 罗祖维
Original assignee: Wuhan Textile University
Current assignee: Wuhan Textile University
Priority date: 2024-01-05
Filing date: 2024-01-05
Publication date: 2024-05-03

Abstract

The invention provides a double-network interpenetrating hydrogel microneedle based on silk fibroin and acylated hyaluronic acid and a preparation method thereof. Tyrosine on a silk fibroin macromolecular chain in SF is oxidized into tyrosine residues under the catalysis of ultraviolet light excitation and riboflavin, the tyrosine residues are crosslinked with each other to form a dityrosine single-network hydrogel, and acetyl-grafted hyaluronic acid is crosslinked with ultraviolet light under the catalysis of a photoinitiator to form a HAMA hydrogel network. The initiation of SF by riboflavin under UV light is an extremely slow process, the formation of gel by HAMA under UV light is a chain reaction, and the photo-curing of both can accelerate the gel rate of SF after mixing. The HAMA single-network hydrogel is used as a drug carrier and has an excessively high release rate in tissue fluid, the defect that HAMA rapidly swells to suddenly release insulin can be alleviated by adding SF, and finally the double-network hydrogel with excellent slow release effect is prepared.

Description

Double-network interpenetrating hydrogel microneedle based on silk fibroin and acylated hyaluronic acid and preparation method thereof

Technical Field

The invention relates to the field of preparation of hydrogel microneedles, in particular to a double-network interpenetrating hydrogel microneedle based on silk fibroin and acylated hyaluronic acid and a preparation method thereof.

Background

Hydrogel microneedles are a new type of microneedle that has been developed in recent years. The solid micro-needle has the characteristics of swelling but not dissolving after penetrating into subcutaneous tissue fluid, can be completely removed from skin after administration, and overcomes the risk of easy breakage of the solid micro-needle in vivo, so that the solid micro-needle is widely developed for sustained release of medicines or extraction of tissue fluid and the like, and the solid micro-needle is widely applied as an insulin medicine carrying platform. Silk fibroin is used as a natural biological protein, and is a preferred material for developing functional biomedical appliances because of the advantages of good biocompatibility, biodegradability, excellent processability, easiness in mass production and the like. Hyaluronic acid, which is a polyanionic disaccharide extracted from vitreous bodies of bulls eye, widely exists in connective tissue and dermis layers of human beings, is one of main components of extracellular matrix of human body, and is excellent in biocompatibility, water-retaining ability and operability, and is considered as an excellent hydrogel preparation material. However, single-network hyaluronic acid hydrogels often have poor mechanical properties, poor structural stability, and difficulty in sustained and controlled drug release, and cannot meet the requirement of hydrogel microneedles as insulin delivery platforms. In recent years, efforts have been made to improve the mechanical properties, structural stability and functional controllability of single network hydrogels, and research methods include developing dual network structures, physicochemical crosslinking, solvent induction, and nanocomposite materials. Among them, the dual-network structure has advantages of structural adjustability, versatility, etc., which makes them one of the preparation modes of materials of great interest in the biomedical field, however, most of the synthesis of dual-network hydrogels requires temperature regulation, photo-thermal, etc. or chemical crosslinking, which may cause denaturation and inactivation of drugs, and the gel curing period is long, which is unfavorable for the rapid preparation and mass production of dual-network gel microneedles. In view of the above, there is a need to design a dual-network interpenetrating hydrogel microneedle based on silk fibroin and acylated hyaluronic acid and a preparation method thereof, so as to solve the above problems.

Disclosure of Invention

Aiming at the defects in the prior art, the invention aims to provide a double-network interpenetrating hydrogel microneedle based on silk fibroin and acylated hyaluronic acid and a preparation method thereof, wherein silk fibroin and acylated hyaluronic acid are used as precursor materials of a two-phase network structure, and the double-network interpenetrating hydrogel microneedle capable of rapidly preparing the silk fibroin/hyaluronic acid groups which are stable in structure and excellent in biocompatibility and can be regulated and controlled is developed, so that the double-network interpenetrating hydrogel microneedle is used as an insulin transport platform, and has great significance in solving clinical challenges facing diabetes.

In order to achieve the above purpose, the invention provides a preparation method of a double-network interpenetrating hydrogel microneedle based on silk fibroin and acylated hyaluronic acid, which comprises the following steps:

s1, preparing regenerated SF solution

Dissolving degummed raw silk fibers in LiBr solution with preset concentration, and dialyzing to obtain SF solution; filtering and centrifuging the SF solution to obtain regenerated SF solution with a preset concentration;

S2, preparation of SF-RF solution

Uniformly mixing the regenerated SF solution with riboflavin according to a preset volume ratio to prepare SF-RF solution;

s3, preparing acylated hyaluronic acid HAMA pre-coagulant

Slowly adding methacrylic anhydride into hyaluronic acid with preset concentration, reacting for preset time, sequentially dialyzing, and freeze-drying to obtain HAMA; uniformly mixing the re-dissolved HAMA with a photoinitiator I2959, and standing to prepare an HAMA prefoagulant;

s4, preparing hydrogel microneedle

Mixing the SF-RF solution with the HAMA pre-coagulant according to a preset volume ratio to prepare a mixed solution; adding human recombinant insulin with preset concentration into the mixed solution to prepare insulin microneedle precursor liquid; then the insulin microneedle precursor liquid is injected into a microneedle mould for vacuumizing treatment, and then is taken out for air drying treatment; then injecting the mixed solution into the microneedle mould for a plurality of times, exposing the mixed solution to ultraviolet rays for a preset time, and carrying out photopolymerization to obtain a photopolymerization gel microneedle; the photopolymerized gel microneedle is then dried to produce a hydrogel microneedle.

Further, in step S1, the degumming process is to put the raw silk fiber into a boiling Na ₂CO₃ solution with a concentration of 0.05 wt%.

Further, in step S1, the predetermined concentration of the LiBr solution is 9.3mol/L.

Further, in step S1, the predetermined concentration of the regenerated SF solution is 5.0wt%.

Further, in step S2, the predetermined volume ratio is: regenerated SF solution: riboflavin=10 to 500:1.

Further, in the step S3, the preset concentration of the hyaluronic acid is 5-150 g/L; the volume ratio of the hyaluronic acid to the methacrylic anhydride is 100:1-10:1; the preset time for carrying out the reaction is 6-24 hours;

The photoinitiator I2959 included 2-hydroxy-4' - (2-hydroxyethyl) -2-methylpropionne at a concentration of 0.15 wt%; the solute ratio of the HAMA to the photoinitiator I2959 is 10-400:1.

Further, in the step S4, the predetermined volume ratio of the SF solution to the HAMA prefoagulant is 95:5-5:95.

Further, in the step S4, the concentration of the human recombinant insulin is 1-50wt%; the solute ratio of the mixed solution to the human recombinant insulin is 1000:1-5:1.

Further, in step S4, the predetermined time of exposure to ultraviolet rays is 1 to 2 minutes.

The invention also provides a double-network interpenetrating hydrogel microneedle based on the silk fibroin and the acylated hyaluronic acid and a hydrogel microneedle prepared by the preparation method thereof; the array of hydrogel microneedles is in an 11 x 11 arrangement; each microneedle has a pyramid shape with a base width of 200-600 μm and a height of 400-800 μm.

The beneficial effects of the invention are as follows:

1. According to the preparation method of the double-network interpenetrating hydrogel microneedle based on silk fibroin and acylated hyaluronic acid, tyrosine on a silk fibroin macromolecular chain in SF is oxidized into tyrosine residues under the catalysis of riboflavin through the excitation of ultraviolet light, the tyrosine residues are crosslinked with each other to form a dityrosine single-network hydrogel, and the grafted acetyl hyaluronic acid can promote HAMA to be rapidly photocrosslinked to form a hydrogel network under the catalysis of a photoinitiator. SF is an extremely slow process in the case of ultraviolet light, while HAMA forms a gel in the case of ultraviolet light, which is a chain reaction, and gel formation is rapid. The HAMA photopolymerization process can accelerate the gel rate of SF, the swelling rate of HAMA single-network hydrogel in tissue fluid is too high, and the addition of SF network can slow down the insulin burst caused by rapid swelling of HAMA. And SF in a dry state has good mechanical properties, the structural stability of the microneedle is enhanced by the formation of interpenetrating hydrogel, and finally the double-network hydrogel microneedle with excellent mechanical properties and slow release characteristics is obtained.

2. The double-network interpenetrating hydrogel microneedle based on the silk fibroin and the acylated hyaluronic acid provided by the invention can realize the mechanical strength required by the penetration of the microneedle into the skin and the structural stability of materials in the action process by taking the double-network hydrogel as a precursor of the microneedle, the microneedle prepared from the double-network hydrogel can swell and release drugs in vivo and can be controlled accurately, the breaking force can reach 0.6N/needle, and the maximum load is 79.3+/-11.2N. The photo-curing double-network swelling drug release microneedle system prepared by the method is simple and efficient in preparation and suitable for industrialized mass production.

Drawings

FIG. 1 is an electron micrograph of the hydrogel microneedle prepared in example 1, wherein (A) is a global morphology map (optical photograph) of the microneedle, (B) is a local bright field map (microscopic bright field) of the microneedle, (C) is a morphology map (SEM) of the microneedle scanning electron microscope image, and (D) is a confocal microscopy schematic of a double network gel structure of the microneedle (red: HAMA network, green: silk fibroin network).

Fig. 2 is a wound healing process formed after the hydrogel microneedle prepared in example 1 is pressed against the skin surface of a human body.

FIG. 3 is a graph showing cytotoxicity assay experiments performed on hydrogel microneedles prepared in example 1, wherein (A) is a live/dead stained image of HUVECs (human umbilical vein endothelial cells) cultured on the surface of the hydrogel microneedles, and (B) is the OD values of HUVECs on the microneedles at various times (1, 3, 7 days).

Fig. 4 shows mechanical property tests of hydrogel microneedles prepared in accordance with examples and comparative examples, wherein (a) is a graph of force versus displacement of breaking compression of each set of microneedles, (B) is a maximum bearing capacity of each set of microneedles, and (C) is a swelling ratio of each set of microneedles.

Detailed Description

In order to make the development objects, technical solutions and advantageous explanation of the present invention more clear, the present invention is described in detail below with reference to the accompanying drawings and specific embodiments.

It should be noted that, in order to avoid obscuring the present invention due to unnecessary details, only structures and/or processing steps closely related to aspects of the present invention are shown in the drawings, and other details not greatly related to the present invention are omitted.

In addition, it should be further noted that the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.

A preparation method of a double-network interpenetrating hydrogel microneedle based on silk fibroin and acylated hyaluronic acid comprises the following steps:

s1, preparing regenerated SF solution

Firstly, putting raw silk fibers into a Na ₂CO₃ solution which is boiled at 100 ℃ and has the concentration of 0.05 weight percent for degumming, then dissolving the raw silk fibers subjected to repeated degumming and water washing into a LiBr solution with the concentration of 9.3mol/L, and then putting into deionized water for dialysis for 3 days to prepare SF solution; filtering and centrifuging the SF solution to obtain a regenerated SF solution with the concentration of 5.0 wt%;

S2, preparation of SF-RF solution

Uniformly mixing the regenerated SF solution and riboflavin according to the volume ratio of 10-500:1 to prepare SF-RF solution;

s3, preparing acylated hyaluronic acid HAMA pre-coagulant

Slowly adding methacrylic anhydride into hyaluronic acid with the concentration of 5-150 g/L, reacting for 6-24 h, sequentially dialyzing in deionized water for 3 days, and freeze-drying to obtain HAMA; uniformly mixing the re-dissolved HAMA with a photoinitiator I2959, and standing for 1-4 h to prepare an HAMA pre-coagulant;

the volume ratio of the hyaluronic acid to the methacrylic anhydride is 100:1-10:1;

The freeze-drying process is as follows: placing the mixed solution in liquid nitrogen at-196 ℃ for 30min, taking out, and placing in a freeze dryer for drying treatment;

The photoinitiator I2959 included 2-hydroxy-4' - (2-hydroxyethyl) -2-methylpropionne at a concentration of 0.15 wt%; the solute ratio of the HAMA to the photoinitiator I2959 is 10-400:1;

s4, preparing hydrogel microneedle

Mixing the SF-RF solution and the HAMA pre-coagulant according to the volume ratio of 95:5-5:95 to prepare a mixed solution; then adding 1-50wt% of human recombinant insulin into the mixed solution to prepare insulin microneedle precursor liquid; injecting the insulin microneedle precursor liquid into a microneedle mould, vacuumizing, and taking out the insulin microneedle precursor liquid subjected to vacuum treatment for air drying; then injecting the mixed solution into the microneedle mould for a plurality of times, and exposing the mixed solution to 365nm Ultraviolet (UV) for 1-2 min for photopolymerization to prepare the photopolymerization gel microneedle; drying the photopolymerization gel microneedle to obtain a hydrogel microneedle;

the ratio of the mixed solution to the human recombinant insulin is 1000:1-5:1.

The setting is that tyrosine on silk fibroin macromolecular chain in SF is oxidized into tyrosine residue under the catalysis of riboflavin by ultraviolet light excitation, the tyrosine residues are cross-linked to form a dityrosine single-network hydrogel, and acetyl-grafted hyaluronic acid can rapidly form HAMA hydrogel network under the catalysis of photoinitiator. The gel of SF is a very slow process under ultraviolet light, meanwhile, the gel formation of HAMA under ultraviolet light is a chain reaction, the gel rate of SF can be accelerated while the gel is fast, the release rate of HAMA single-network hydrogel in tissue fluid is too fast, the defect that the HAMA swells fast and releases insulin suddenly can be alleviated by adding SF network, and SF in a dry state has good mechanical property, so that the double-network hydrogel with excellent mechanical property can be finally prepared. Wherein, when preparing hydrogel microneedles, if the concentration of the added insulin is too high, the insulin can be suddenly released; too low a concentration of insulin added may result in too slow insulin release.

The invention also provides a hydrogel microneedle prepared by the preparation method of the double-network interpenetrating hydrogel microneedle based on silk fibroin and acylated hyaluronic acid; the array of hydrogel microneedles is in an 11 x 11 arrangement; each microneedle has a pyramid shape with a base width of 200-600 μm and a height of 400-800 μm.

By the arrangement, the double-network hydrogel is used as a precursor of the microneedle, can provide mechanical performance support before the microneedle is inserted into the skin, has strong control over the swelling and drug release of the microneedle prepared from the double-network hydrogel in vivo, and has a fast process.

The following specifically describes a silk fibroin and acylated hyaluronic acid-based dual-network interpenetrating hydrogel microneedle and a preparation method thereof, which are provided by the invention, by combining with examples:

example 1

The embodiment provides a double-network interpenetrating hydrogel microneedle based on silk fibroin and acylated hyaluronic acid and a preparation method thereof, and the preparation method specifically comprises the following steps:

s1, preparing regenerated SF solution

Firstly, degumming raw silk fiber in Na ₂CO₃ solution boiling at 100 ℃ and with the concentration of 0.05wt% for 30min, dissolving the raw silk fiber after 3 times of degumming water washing in LiBr solution with the concentration of 9.3mol/L, and then, dialyzing in deionized water for 3 days to obtain SF solution; filtering and centrifuging the SF solution to obtain a regenerated SF solution with the concentration of 5.0 wt%;

the degumming process is to put raw silk fiber into boiling Na ₂CO₃ solution with concentration of 0.05wt percent for degumming;

S2, preparation of SF-RF solution

100ML of the regenerated SF solution and 1mL of riboflavin (10 mmol/L, china biological organisms) are uniformly mixed to prepare SF-RF solution;

s3, preparing HAMA pregelatinization agent

Slowly adding 6mL of methacrylic anhydride into a hyaluronic acid aqueous solution with the concentration of 10g/L, reacting for 12 hours, sequentially dialyzing in deionized water for 3 days, and freeze-drying to obtain HAMA; uniformly mixing the re-dissolved HAMA with a photoinitiator I2959 of 2-hydroxy-4' - (2-hydroxyethyl) -2-methyl propiophenone with the concentration of 0.15wt%, and standing for 3 hours to prepare an HAMA prefoaming agent;

The volume ratio of the hyaluronic acid to the methacrylic anhydride is 100:3;

the solute ratio of the HAMA to the photoinitiator I2959 is 40:3;

s4, preparing hydrogel microneedle

Mixing the SF-RF solution and the HAMA pre-coagulant according to the volume ratio of 50:50 to prepare a mixed solution; adding 5wt% dry human recombinant insulin into the mixed solution to prepare insulin microneedle precursor liquid; injecting the insulin microneedle precursor liquid into a microneedle mould, performing vacuumizing program treatment for 5min, taking out the insulin microneedle precursor liquid subjected to vacuum treatment, and performing air drying treatment at room temperature; then injecting the mixed solution into the microneedle mould for multiple times, and exposing the mixed solution to 365nm Ultraviolet (UV) for 2min for photopolymerization to prepare a photopolymerization gel microneedle; drying the photopolymerization gel microneedle to obtain a hydrogel microneedle; the hydrogel only needs to be formed for 90 seconds;

The ratio of the mixed solution to the human recombinant insulin is 20:1.

In addition, as a result of observing the microstructure of the hydrogel microneedle prepared in this example, as shown in fig. 1, it was seen in fig. 1 (a to C) that the microneedle was pyramid-shaped and smooth in surface, and an 11×11 microneedle array having a height of about 400 to 800 μm and a bottom width of 200 to 600 μm was formed, and the height of the microneedle was sufficient to reach the dermis layer of skin tissue to achieve insulin delivery. As can be seen from FIG. 1 (D), the SF network structure dyed with the ThT DYE into green fluorescence and the HAMA network dyed with the DYE-R DYE into red fluorescence can observe clear double network structures under a confocal microscope, and the networks formed by the SF network structure and the HAMA network structure uniformly interpenetrate each other without interfering with each other, so that the network structure is uniform, high-density and ordered. Fig. 2 is a graph of a wound healing process formed after the microneedle prepared in the embodiment is pressed on the surface of human skin for 3min, and a clear micro-wound in the shape of a microneedle array can be observed after the microneedle is pulled out from the skin, and after 30min, the surface of the skin is completely recovered, so that the requirement of micro-invasive drug administration of the microneedle is met.

Meanwhile, the hydrogel microneedles prepared in this example were also subjected to biocompatibility test, as shown in fig. 3, human umbilical vein endothelial cells (hUVECs) were respectively cultured on the microneedle patches, live/dead fluorescent staining images of HUVECs growing for 1d and 7d on each group of microneedles are shown in fig. 3 (a), HUVECs are paving stones, the filiform pseudopodites of HUVECs on the microneedles are observed in the cytostaining images to adhere to the surfaces of the microneedles, rapid expansion is performed along the needle columns towards the needle tip, and it is shown that each group of microneedles can support adhesion and proliferation of cells, and the Live/dead fluorescent staining images show HUVECs (apoptotic cells show red fluorescence, living cells show green fluorescence) hardly observed on the surfaces of the microneedles, so that the HUVECs have good growth state. The surface of the SF/HAMA microneedle is completely covered by cells after being cultured for 7d, and the HUVECs grow into a network structure, so that the phenomenon is that the hydrogel micro-particles have a certain adhesion effect on the HUVECs, and meanwhile, the HUVECs are not easy to slide down onto a microneedle patch base when growing on the microneedle. As shown in FIG. 3 (B), HUVECs are inoculated on the microneedles, and cell viability detection shows that the HUVECs can be successfully inoculated on each group of the microneedles, and the cell number is obviously increased along with the prolonged culture time; the OD of the microneedles increased significantly when cultured to 3d and 7d, indicating that HUVECs can proliferate rapidly on the microneedles; and the SF/HAMA microneedle has stable structure, and based on the advantages, the cells on the SF/HAMA microneedle can continuously and rapidly grow, and the proliferation activity of the cells on the SF/HAMA microneedle completely meets the requirement of insulin micro-alignment cell compatibility. Each microneedle had a pyramid shape with a base width of 400 μm and a height of 700 μm.

Examples 2 to 3

Examples 2 to 3 respectively provide a dual-network interpenetrating hydrogel microneedle based on silk fibroin and acylated hyaluronic acid and a preparation method thereof, and compared with example 1, the difference is that in step S4, the volume ratio of SF solution to HAMA prefoaming agent is different: the volume ratio of SF solution to HAMA prefoagulator in example 2 was 80:20; the volume ratio of SF solution to HAMA prefoagulator in example 3 was 20:80. The remaining steps and parameters are the same as those of embodiment 1, and will not be described again.

Example 4

Example 4 provides a silk fibroin and acylated hyaluronic acid-based dual network interpenetrating hydrogel microneedle and a preparation method thereof, which are different from example 1 in that in step S4, the concentration of added human recombinant insulin is different: example 4 the concentration of human recombinant insulin added was 20wt%. The remaining steps and parameters are the same as those of embodiment 1, and will not be described again.

Comparative examples 1 to 2

Comparative examples 1 to 2 respectively provide a silk fibroin and acylated hyaluronic acid-based double-network interpenetrating hydrogel microneedle and a preparation method thereof, which are different from example 1 in that in step S4, the volume ratio of SF solution to HAMA prefoaming agent is different: the volume ratio of SF solution to HAMA prefoagulator in comparative example 1 is 100:0; the volume ratio of SF solution to HAMA prefoagulator in comparative example 2 was 0:100. The remaining steps and parameters are the same as those of embodiment 1, and will not be described again.

Comparative example 3

Comparative example 3 provides a double-network interpenetrating hydrogel microneedle based on silk fibroin and acylated hyaluronic acid and a preparation method thereof, which is different from example 1 in that the concentration of added human recombinant insulin is different in step S4: comparative example 3 the concentration of human recombinant insulin added was 60wt%. The encapsulation efficiency of the finally prepared hydrogel microneedle is reduced, so that the medicine in the hydrogel microneedle can not be fully injected into a human body, and waste is caused.

Subsequently, mechanical tests were performed on the individual microneedles of the hydrogel microneedles prepared in examples 1 to 4 and comparative examples 1 to 2, respectively, and as shown in fig. 4, it was found that the needles prepared with a volume ratio of SF solution to HAMA prefoaming of 0:100 had more than 0.4N/needle except that the breaking force of the needles did not exceed 0.4N/needle; however, when the volume ratio of SF solution to HAMA pre-coagulant is 100:0, the prepared microneedle can be gradually dissolved in the swelling process, water is not absorbed and swelled, gel cannot be formed, and finally the prepared microneedle cannot be molded; thus, when the volume ratio of SF solution to HAMA prefoagulator is in the range of 95:5-5:95, the breaking force of the prepared micropins can reach as high as 0.6N/needle, and the maximum load is 79.3+/-11.2N, so that the micropins can penetrate into dermis of skin for administration.

In summary, according to the double-network interpenetrating hydrogel microneedle based on silk fibroin and acylated hyaluronic acid and the preparation method thereof provided by the invention, tyrosine on a silk fibroin macromolecular chain in SF is oxidized into tyrosine residues under the catalysis of riboflavin, the tyrosine residues are crosslinked with each other to form a double-tyrosine single-network hydrogel, and the hyaluronic acid grafted with acetyl can rapidly form a HAMA hydrogel network under the catalysis of a photoinitiator. The SF is a very slow process under ultraviolet light, meanwhile, the HAMA forms gel under ultraviolet light is a chain reaction, the gel rate of SF can be accelerated while the gel is fast, the release rate of HAMA single-network hydrogel in tissue fluid is too fast, the defect that the HAMA rapidly swells and cannot slowly release insulin can be overcome by adding an SF network, the SF under a dry state has good mechanical properties, and finally, the double-network hydrogel microneedle with excellent mechanical properties can be prepared.

The above embodiments are only for illustrating the technical solution of the present invention and not for limiting the same, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications and equivalents may be made thereto without departing from the spirit and scope of the technical solution of the present invention.

Claims

1. The preparation method of the double-network interpenetrating hydrogel microneedle based on silk fibroin and acylated hyaluronic acid is characterized by comprising the following steps of:

s1, preparing regenerated SF solution

S2, preparation of SF-RF solution

s3, preparing acylated hyaluronic acid HAMA pre-coagulant

s4, preparing hydrogel microneedle

2. The method for preparing the silk fibroin and acylated hyaluronic acid-based double-network interpenetrating hydrogel microneedle according to claim 1, wherein the method comprises the following steps: in step S1, the degumming process is to put raw silk fiber into boiling Na ₂CO₃ solution with concentration of 0.05 wt%.

3. The method for preparing the silk fibroin and acylated hyaluronic acid-based double-network interpenetrating hydrogel microneedle according to claim 1, wherein the method comprises the following steps: in step S1, the predetermined concentration of the LiBr solution is 9.3mol/L.

4. The method for preparing the silk fibroin and acylated hyaluronic acid-based double-network interpenetrating hydrogel microneedle according to claim 1, wherein the method comprises the following steps: in step S1, the predetermined concentration of the regenerated SF solution is 5.0wt%.

5. The method for preparing the silk fibroin and acylated hyaluronic acid-based double-network interpenetrating hydrogel microneedle according to claim 1, wherein the method comprises the following steps: in step S2, the predetermined volume ratio is: regenerated SF solution: riboflavin=10 to 500:1.

6. The method for preparing the silk fibroin and acylated hyaluronic acid-based double-network interpenetrating hydrogel microneedle according to claim 1, wherein the method comprises the following steps: in the step S3, the preset concentration of the hyaluronic acid is 5-150 g/L; the volume ratio of the hyaluronic acid to the methacrylic anhydride is 100:1-10:1; the preset time for carrying out the reaction is 6-24 hours;

7. The method for preparing the silk fibroin and acylated hyaluronic acid-based double-network interpenetrating hydrogel microneedle according to claim 1, wherein the method comprises the following steps: in the step S4, the preset volume ratio of the SF solution to the HAMA prefoaming agent is 95:5-5:95.

8. The method for preparing the silk fibroin and acylated hyaluronic acid-based double-network interpenetrating hydrogel microneedle according to claim 1, wherein the method comprises the following steps: in the step S4, the concentration of the human recombinant insulin is 1-50wt%; the ratio of the mixed solution to the human recombinant insulin is 1000:1-5:1.

9. The method for preparing the silk fibroin and acylated hyaluronic acid-based double-network interpenetrating hydrogel microneedle according to claim 1, wherein the method comprises the following steps: in step S4, the predetermined time of exposure to ultraviolet rays is 1 to 2 minutes.

10. A silk fibroin and acylated hyaluronic acid-based dual-network interpenetrating hydrogel microneedle, characterized in that: the hydrogel microneedle prepared according to the preparation method of any one of claims 1 to 9; the array of hydrogel microneedles is in an11 x 11 arrangement; each microneedle has a pyramid shape with a base width of 200-600 μm and a height of 400-800 μm.