Silk fibroin hyaluronic acid double-network gel for filling nasolabial sulcus and preparation process of silk fibroin hyaluronic acid double-network gel

Abstract

The invention relates to the technical field of polymer gel materials, in particular to silk fibroin hyaluronic acid double-network gel for filling nasolabial grooves and a preparation process of the silk fibroin hyaluronic acid double-network gel. According to the invention, the tannic acid, the cinnamyl aldehyde, the lidocaine and the mesoporous polydopamine nanoparticles are added, so that the composite antibacterial material is prepared. The preparation method comprises the following steps: adding hyaluronic acid, sodium periodate, propylated chitosan and a photoinitiator 2959 to obtain a solution A; adding silk and sodium alginate to obtain a solution B; mixing the solution A and the solution B, fully stirring, adding a composite antibacterial material, and continuously stirring to obtain a mixed cross-linking solution; sequentially carrying out ultrasonic defoaming, freezing overnight and freeze drying on the mixed cross-linking solution to obtain aerogel; and dipping the aerogel in a calcium chloride solution, and carrying out ultraviolet crosslinking, water washing and freeze drying to obtain a finished product. The finished product prepared by the invention has excellent antibacterial property and high crosslinking degree, so that the finished product has wide application prospects in the technical field of polymer gel materials.

Description

Technical Field

[0001] The present invention relates to the technical field of polymer gel materials, and specifically to a fibroin hyaluronic acid double-network gel for nasolabial fold filling and its preparation process. Background Art

[0002] With the rapid development of the medical aesthetics industry, injectable soft tissue fillers have received extensive attention. Among them, fibroin and sodium hyaluronate, as natural polymer materials, have good biocompatibility and are used in multiple fields such as tissue engineering and drug sustained release, with broad market application prospects. Although sodium hyaluronate gel has good immediate filling and shaping effects, it degrades rapidly in the tissue environment, has a low hardness, does not have the function of tissue regeneration, and patients need to be injected frequently. Fibroin is a fibrous protein with a slow biodegradation rate, which has advantages in slow degradation. At the same time, it has strong adhesion and guidance migration ability to cells and can provide a growth environment for cells. However, the water retention and compressive properties of regenerated fibroin gel need to be improved. Therefore, crosslinking the two to form a composite gel can combine the advantages of both, extend the duration of the filling effect, improve the effectiveness of the material, and meet the requirements of soft tissue repair for the material.

[0003] Defects and deficiencies of the prior art:

[0004] 1. Poor network structure stability: If the network inside the gel is formed by the mixed crosslinking of fibroin and sodium hyaluronate, when the sodium hyaluronate component is degraded first, it is easy to cause the collapse and disintegration of the overall three-dimensional network of the gel, making the gel lose its effectiveness and unable to achieve the purpose of stable support and filling in the body.

[0005] 2. Prone to bacterial contamination: In practical applications, especially when used for human tissue filling, the gel faces the risk of infection. Once the gel is contaminated by bacteria, it may cause local inflammatory reactions such as redness, swelling, pain, and fever, seriously affecting the filling effect and the health of patients. Improving the antibacterial performance can effectively inhibit the growth and reproduction of bacteria and reduce the probability of infection.

[0006] 3. Affected injectability and lubricity: The crosslinking degree of pure fibroin-sodium hyaluronate crosslinked gel is low, and its mechanical properties are poor, with low elasticity and high viscosity, which will affect the injectability and lubricity of the gel. If the crosslinking of the gel is insufficient and the mechanical strength is low, it is easy to deform, shift, or even break, resulting in an unsatisfactory filling effect and affecting facial aesthetics.

[0007] 4. Difficult to control the reaction: The preparation method of fibroin-sodium hyaluronate crosslinked double-network gel is complex. It is difficult to achieve the crosslinking of fibroin and the preparation of gel in an aqueous solution under normal alkaline reaction conditions. When using multiple crosslinking agents, the reaction process is difficult to precisely control, which may lead to unstable gel properties.

[0008] To overcome the defects of the prior art, the present invention provides a silk fibroin - hyaluronic acid double - network gel for nasolabial fold filling and its preparation process. Summary of the Invention

[0009] The object of the present invention is to provide a silk fibroin - hyaluronic acid double - network gel for nasolabial fold filling and its preparation process to solve the problems raised in the prior art.

[0010] To achieve the above object, the present invention provides the following technical solutions:

[0011] The preparation process of the silk fibroin - hyaluronic acid double - network gel for nasolabial fold filling includes the following steps:

[0012] Step 1: Add tannic acid, cinnamaldehyde, and lidocaine to an anhydrous ethanol solution, stir well for 5 - 8 min to obtain a reaction solution; disperse mesoporous polydopamine nanoparticles into the reaction solution, react at 25 - 30 °C in the dark for 25 - 30 h, and then perform centrifugation, washing, and freeze - drying to obtain a composite antibacterial material;

[0013] Step 2: Dissolve hyaluronic acid in deionized water, then add sodium periodate, react at 25 - 30 °C in the dark for 25 - 30 h, and after the reaction, perform dialysis and freeze - drying to obtain oxidized hyaluronic acid; dissolve the oxidized hyaluronic acid in deionized water to obtain an oxidized hyaluronic acid solution; add acrylated chitosan and photoinitiator 2959 to an ethanol - aqueous solution, stir well for 10 - 20 min to obtain a chitosan solution; then mix the oxidized hyaluronic acid solution and the chitosan solution, stir well for 10 - 20 min to obtain solution A;

[0014] Degum, dissolve, dialyze, and dissolve in deionized water the silk to obtain a silk fibroin solution, then add sodium alginate, stir well until dissolved to obtain solution B; mix solution A and solution B, stir well for 3 - 5 min, then add the composite antibacterial material, and continue to stir for 15 - 20 min to obtain a mixed cross - linked solution;

[0015] Step 3: Perform ultrasonic degassing, freeze overnight, and freeze - dry the mixed cross - linked solution in sequence to obtain an aerogel; then immerse the aerogel in a calcium chloride solution, perform photocross - linking under ultraviolet light conditions for 3 - 5 min, and then perform water washing and freeze - drying to obtain the finished product.

[0016] Preferably, in step 1, the concentration of tannic acid in the reaction solution is 1.8 - 2.0 mg / mL, the concentration of cinnamaldehyde in the reaction solution is 0.5 - 0.7 mg / mL, the concentration of lidocaine in the reaction solution is 1.0 - 1.5 mg / mL, and the concentration of mesoporous polydopamine nanoparticles in the reaction solution is

[0017] 3.0 - 3.5 mg / mL.

[0018] Preferably, in step one, the preparation process of the mesoporous polydopamine nanoparticles is as follows: Pluronic F-127 and 1,3,5-trimethylbenzene are sequentially added to an ethanol aqueous solution, and after ultrasonic dispersion to obtain a white emulsion, magnetic stirring is carried out for 30 - 40 min. Then, a Tris-HCl solution and dopamine hydrochloride are sequentially added, the pH is adjusted to 8.5 - 8.7, and a reaction is carried out at 25 - 30 °C in the dark for 25 - 30 h. After the reaction is completed, solid nanoparticles are collected at 3 - 5 °C, and then through ultrasonic cleaning, centrifugal collection, washing with deionized water, and freeze-drying, mesoporous polydopamine nanoparticles are obtained.

[0019] Preferably, the reaction mass ratio of Pluronic F-127, 1,3,5-trimethylbenzene, Tris-HCl, and dopamine hydrochloride is 37:36:9:(6.0 - 6.5).

[0020] Preferably, in step two, the preparation process of acrylated chitosan is as follows: Chitosan is dissolved in a 2.0 - 2.5 wt% acetic acid solution, stirred at 25 - 30 °C for 10 - 12 h, then methacrylic anhydride is added dropwise and a reaction is carried out in a water bath at 60 - 65 °C for 6 - 8 h. After the reaction is completed, the pH is adjusted to neutral, dialysis is carried out, and freeze-drying is carried out to obtain acrylated chitosan.

[0021] Preferably, the mass-volume ratio of chitosan, acetic acid solution, and methacrylic anhydride is 1 g:50 mL:(2.0 - 2.5) mL.

[0022] Preferably, in step two, when preparing oxidized hyaluronic acid, the reaction mass ratio of hyaluronic acid and sodium periodate is (0.9 - 1.0):1.

[0023] Preferably, in step two, when preparing solution A, the reaction mass ratio of oxidized hyaluronic acid, acrylated chitosan, and photoinitiator 2959 is 1:(0.7 - 0.9):0.02; when preparing solution B, the reaction mass ratio of sodium alginate and silk fibroin is 3:(3 - 4); when preparing the mixed cross-linking solution, the reaction mass ratio of oxidized hyaluronic acid, silk fibroin, and the composite antibacterial material is (1.2 - 1.5):3:(0.3 - 0.5).

[0024] Preferably, in step three, the calcium chloride solution is 1.5 - 2.0 wt%; the ultraviolet light intensity is 25 - 35 mW / cm 2 .

[0025] The beneficial effects of the present invention:

[0026] The characteristics of the present invention are as follows. In Step 1, a mesoporous polydopamine nanoparticle is prepared by adding Pluronic F-127, 1,3,5-trimethylbenzene, Tris-HCl, and dopamine hydrochloride. Then, tannic acid, cinnamaldehyde, and lidocaine are loaded into the pores of the mesoporous polydopamine nanoparticle to obtain a composite antibacterial material. The polydopamine in this composite antibacterial material itself has good biocompatibility and can interact well with biological tissues and cells, reducing irritation and immune responses to organisms. In addition, the mesoporous polydopamine nanoparticle also has excellent photothermal conversion ability and can convert light energy into heat energy under the irradiation of near-infrared light, generating local high temperature. This photothermal effect can not only directly kill bacteria but also promote the release of the loaded drug and enhance the antibacterial effect. Tannic acid and cinnamaldehyde both have certain antibacterial activities and act on different targets of bacteria. Tannic acid can bind to the proteins on the surface of bacteria and destroy the cell wall and cell membrane structures of bacteria; cinnamaldehyde can inhibit the respiratory chain and enzyme activities of bacteria and interfere with the metabolic process of bacteria. The synergistic effect of the two can expand the antibacterial spectrum, improve the antibacterial efficiency, and have good inhibitory effects on a variety of Gram-positive and Gram-negative bacteria. Lidocaine is a commonly used local anesthetic, and by being loaded in the composite antibacterial material, it can provide analgesic effects while antibacterial.

[0027] The characteristics of the present invention are as follows. In Step 2 and Step 3, hyaluronic acid is oxidized by sodium periodate. The oxidized hyaluronic acid contains more aldehyde groups, which provides active sites for subsequent cross-linking reactions with chitosan. The oxidized hyaluronic acid solution and the chitosan solution are mixed and stirred, and the two can cross-link through the reaction between aldehyde groups and amino groups to form a preliminary network structure, obtaining Solution A. Sodium alginate is added to the aqueous solution of silk fibroin and stirred until dissolved to obtain Solution B. Silk fibroin and sodium alginate have good biocompatibility and certain gel-forming properties. Solution A, Solution B, and the composite antibacterial material are mixed and then impregnated in a calcium chloride solution. Calcium chloride can undergo ionic cross-linking with sodium alginate. By integrating the properties of various materials, better antibacterial and mechanical properties can be imparted to the gel. Further, under ultraviolet light conditions, photo-cross-linking is carried out for 3 - 5 minutes, and the photoinitiator can initiate the cross-linking reaction of acrylated chitosan, thereby making the structure of the aerogel more stable.

[0028] In summary, the finished product prepared by the present invention has excellent antibacterial properties and high cross-linking degree, so it has broad application prospects in the technical field of polymer gel materials. Detailed implementation manners

[0029] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0030] Source of raw materials:

[0031] Pluronic F-127, provided by Sigma-Aldrich; chitosan, Mw = 200 kDa, degree of deacetylation 85%; hyaluronic acid, provided by Jining Tangyi Chemical Co., Ltd., model number 000; silk, provided by Hai'an Suhao Silk Co., Ltd.; sodium alginate, provided by Anhui Zhonghong Bio-Engineering Co., Ltd., particle size 80 mesh.

[0032] Example 1: Step 1: Add Pluronic F-127 and 1,3,5-trimethylbenzene to an ethanol aqueous solution in sequence, ultrasonically disperse to obtain a white emulsion, then magnetically stir for 40 min, then add Tris-HCl solution and dopamine hydrochloride in sequence, adjust the pH to 8.7, react at 30 °C in the dark for 30 h, after the reaction, collect solid nanoparticles at 5 °C, and then through ultrasonic cleaning, centrifugal collection, washing with deionized water, and freeze-drying to obtain mesoporous polydopamine nanoparticles; the reaction mass ratio of Pluronic F-127, 1,3,5-trimethylbenzene, Tris-HCl, and dopamine hydrochloride is 37:36:9:6.3;

[0033] Add tannic acid, cinnamaldehyde, and lidocaine to an anhydrous ethanol solution, stir well for 8 min to obtain a reaction solution; disperse the mesoporous polydopamine nanoparticles into the reaction solution, react at 30 °C in the dark for 30 h, and then through centrifugation, washing, and freeze-drying to obtain a composite antibacterial material; the concentration of tannic acid in the reaction solution is 1.9 mg / mL, the concentration of cinnamaldehyde in the reaction solution is 0.6 mg / mL, the concentration of lidocaine in the reaction solution is 1.3 mg / mL, and the concentration of mesoporous polydopamine nanoparticles in the reaction solution is 3.2 mg / mL;

[0034] Step 2: Dissolve chitosan in a 2.3 wt% acetic acid solution, stir at 30 °C for 12 h, then dropwise add methacrylic anhydride and react in a water bath at 65 °C for 8 h. After the reaction, adjust the pH to neutral, dialyze, and freeze-dry to obtain acrylated chitosan; the mass-volume ratio of chitosan, acetic acid solution, and methacrylic anhydride is 1 g:50 mL:2.3 mL;

[0035] Dissolve hyaluronic acid in deionized water, then add sodium periodate, and react at 30 °C in the dark for 30 h. After the reaction, dialyze and freeze-dry to obtain oxidized hyaluronic acid; dissolve the oxidized hyaluronic acid in deionized water to obtain an oxidized hyaluronic acid solution; add acrylated chitosan and photoinitiator 2959 to an aqueous ethanol solution, and stir well for 20 min to obtain a chitosan solution; then mix the oxidized hyaluronic acid solution and the chitosan solution, and stir well for 20 min to obtain Solution A; when preparing the oxidized hyaluronic acid, the reaction mass ratio of hyaluronic acid to sodium periodate is 0.95:1;

[0036] Degum, dissolve, dialyze, and dissolve silk in deionized water to obtain a silk fibroin solution, then add sodium alginate, and stir well until dissolved to obtain Solution B; mix Solution A and Solution B, stir well for 5 min, then add a composite antibacterial material, and continue to stir for 20 min to obtain a mixed cross-linking solution; when preparing Solution A, the reaction mass ratio of oxidized hyaluronic acid, acrylated chitosan, and photoinitiator 2959 is 1:0.8:0.02; when preparing Solution B, the reaction mass ratio of sodium alginate to silk fibroin is 3:3.5; when preparing the mixed cross-linking solution, the reaction mass ratio of oxidized hyaluronic acid, silk fibroin, and composite antibacterial material is 1.3:3:0.4;

[0037] Step 3: Perform ultrasonic degassing, freeze overnight, and freeze-dry the mixed cross-linking solution in sequence to obtain an aerogel; then immerse the aerogel in a calcium chloride solution, perform photocrosslinking under ultraviolet light conditions for 5 min, then wash with water and freeze-dry to obtain the finished product; the calcium chloride solution is 1.7 wt%; the ultraviolet light intensity is 35 mW / cm 2 。

[0038] Example 2: Step 1: Add Pluronic F-127 and 1,3,5-trimethylbenzene to an aqueous ethanol solution in sequence. After ultrasonic dispersion to obtain a white emulsion, stir magnetically for 35 min, then add a Tris-HCl solution and dopamine hydrochloride in sequence, adjust the pH to 8.6, and react at 27 °C in the dark for 27 h. After the reaction, collect solid nanoparticles at 4 °C, then perform ultrasonic cleaning, centrifugal collection, washing with deionized water, and freeze-dry to obtain mesoporous polydopamine nanoparticles; the reaction mass ratio of Pluronic F-127, 1,3,5-trimethylbenzene, Tris-HCl, and dopamine hydrochloride is 37:36:9:6.3;

[0039] Tannic acid, cinnamaldehyde, and lidocaine were added to an anhydrous ethanol solution, and stirred thoroughly for 7 min to obtain a reaction solution; mesoporous polydopamine nanoparticles were dispersed in the reaction solution, and reacted at 27 °C in the dark for 27 h, followed by centrifugation, washing, and freeze-drying to obtain a composite antibacterial material; the concentration of tannic acid in the reaction solution was 1.9 mg / mL, the concentration of cinnamaldehyde in the reaction solution was 0.6 mg / mL, the concentration of lidocaine in the reaction solution was 1.3 mg / mL, and the concentration of mesoporous polydopamine nanoparticles in the reaction solution was 3.2 mg / mL;

[0040] Step 2: Chitosan was dissolved in a 2.3 wt% acetic acid solution, stirred at 27 °C for 11 h, then methacrylic anhydride was added dropwise and reacted in a water bath at 62 °C for 7 h. After the reaction, the pH was adjusted to neutral, dialyzed, and freeze-dried to obtain acrylated chitosan; the mass-volume ratio of chitosan, acetic acid solution, and methacrylic anhydride was 1 g:50 mL:2.3 mL;

[0041] Hyaluronic acid was dissolved in deionized water, and sodium periodate was added, and reacted at 27 °C in the dark for 27 h. After the reaction, it was dialyzed and freeze-dried to obtain oxidized hyaluronic acid; the oxidized hyaluronic acid was dissolved in deionized water to obtain an oxidized hyaluronic acid solution; acrylated chitosan and photoinitiator 2959 were added to an ethanol aqueous solution, and stirred thoroughly for 15 min to obtain a chitosan solution; then the oxidized hyaluronic acid solution and the chitosan solution were mixed and stirred thoroughly for 15 min to obtain solution A; when preparing oxidized hyaluronic acid, the reaction mass ratio of hyaluronic acid to sodium periodate was 0.95:1;

[0042] Silk was degummed, dissolved, dialyzed, and dissolved in deionized water to obtain a silk fibroin solution, and then sodium alginate was added and stirred until dissolved to obtain solution B; solution A and solution B were mixed and stirred thoroughly for 4 min, and then the composite antibacterial material was added and stirred continuously for 17 min to obtain a mixed cross-linking solution; when preparing solution A, the reaction mass ratio of oxidized hyaluronic acid, acrylated chitosan, and photoinitiator 2959 was 1:0.8:0.02; when preparing solution B, the reaction mass ratio of sodium alginate to silk fibroin was 3:3.5; when preparing the mixed cross-linking solution, the reaction mass ratio of oxidized hyaluronic acid, silk fibroin, and the composite antibacterial material was 1.3:3:0.4;

[0043] Step 3: The mixed cross-linking solution was successively subjected to ultrasonic degassing, freezing overnight, and freeze-drying to obtain an aerogel; then the aerogel was impregnated in a calcium chloride solution and photo-cross-linked under ultraviolet light conditions for 4 min, followed by washing with water and freeze-drying to obtain the finished product; the calcium chloride solution was 1.7 wt%; the ultraviolet light intensity was 30 mW / cm 2 。

[0044] Example 3: Step 1: Add Pluronic F-127 and 1,3,5-trimethylbenzene to the ethanol aqueous solution in sequence. After ultrasonic dispersion to obtain a white emulsion, stir magnetically for 30 min, then add Tris-HCl solution and dopamine hydrochloride in sequence, adjust the pH to 8.5, react at 25 °C in the dark for 25 h. After the reaction, collect the solid nanoparticles at 3 °C, and then through ultrasonic cleaning, centrifugal collection, washing with deionized water, and freeze-drying to obtain mesoporous polydopamine nanoparticles; the reaction mass ratio of Pluronic F-127, 1,3,5-trimethylbenzene, Tris-HCl, and dopamine hydrochloride is 37:36:9:6.3;

[0045] Add tannic acid, cinnamaldehyde, and lidocaine to the anhydrous ethanol solution, stir well for 5 min to obtain a reaction solution; disperse the mesoporous polydopamine nanoparticles in the reaction solution, react at 25 °C in the dark for 25 h, and then through centrifugation, washing, and freeze-drying to obtain a composite antibacterial material; the concentration of tannic acid in the reaction solution is 1.9 mg / mL, the concentration of cinnamaldehyde in the reaction solution is 0.6 mg / mL, the concentration of lidocaine in the reaction solution is 1.3 mg / mL, and the concentration of mesoporous polydopamine nanoparticles in the reaction solution is 3.2 mg / mL;

[0046] Step 2: Dissolve chitosan in 2.3 wt% acetic acid solution, stir at 25 °C for 10 h, then dropwise add methacrylic anhydride and react in a water bath at 60 °C for 6 h. After the reaction, adjust the pH to neutral, dialyze, and freeze-dry to obtain acrylated chitosan; the mass-volume ratio of chitosan, acetic acid solution, and methacrylic anhydride is 1 g:50 mL:2.3 mL;

[0047] Dissolve hyaluronic acid in deionized water, then add sodium periodate, react at 25 °C in the dark for 25 h. After the reaction, dialyze and freeze-dry to obtain oxidized hyaluronic acid; dissolve the oxidized hyaluronic acid in deionized water to obtain an oxidized hyaluronic acid solution; add acrylated chitosan and photoinitiator 2959 to the ethanol aqueous solution, stir well for 10 min to obtain a chitosan solution; then mix the oxidized hyaluronic acid solution and the chitosan solution, stir well for 10 min to obtain solution A; when preparing oxidized hyaluronic acid, the reaction mass ratio of hyaluronic acid and sodium periodate is 0.95:1;

[0048] Degum, dissolve, dialyze, and dissolve in deionized water the silk to obtain a silk fibroin solution. Then add sodium alginate and stir well until dissolved to obtain Solution B; mix Solution A and Solution B, stir well for 3 min, then add a composite antibacterial material and continue stirring for 15 min to obtain a mixed crosslinking solution; when preparing Solution A, the reaction mass ratio of oxidized hyaluronic acid, acrylated chitosan, and photoinitiator 2959 is 1:0.8:0.02; when preparing Solution B, the reaction mass ratio of sodium alginate to silk fibroin is 3:3.5; when preparing the mixed crosslinking solution, the reaction mass ratio of oxidized hyaluronic acid, silk fibroin, and the composite antibacterial material is 1.3:3:0.4;

[0049] Step 3: Subject the mixed crosslinking solution to ultrasonic degassing, freeze overnight, and freeze-dry to obtain an aerogel; then immerse the aerogel in a calcium chloride solution and perform photocrosslinking under ultraviolet light conditions for 3 min, followed by washing with water and freeze-drying to obtain the finished product; the calcium chloride solution is 1.7 wt%; the ultraviolet light intensity is 25 mW / cm 2 。

[0050] Comparative Example 1: Remove the composite antibacterial material, and the rest is the same as in Example 1. The specific steps are as follows: Step 1: Dissolve chitosan in a 2.3 wt% acetic acid solution, stir at 30°C for 12 h, then add methacrylic anhydride and react in a water bath at 65°C for 8 h. After the reaction, adjust the pH to neutral, dialyze, and freeze-dry to obtain acrylated chitosan; the mass-volume ratio of chitosan, acetic acid solution, and methacrylic anhydride is 1 g:50 mL:2.3 mL;

[0051] Dissolve hyaluronic acid in deionized water, then add sodium periodate and react in the dark at 30°C for 30 h. After the reaction, dialyze and freeze-dry to obtain oxidized hyaluronic acid; dissolve the oxidized hyaluronic acid in deionized water to obtain an oxidized hyaluronic acid solution; add acrylated chitosan and photoinitiator 2959 to an ethanol aqueous solution and stir well for 20 min to obtain a chitosan solution; then mix the oxidized hyaluronic acid solution and the chitosan solution and stir well for 20 min to obtain Solution A; when preparing oxidized hyaluronic acid, the reaction mass ratio of hyaluronic acid to sodium periodate is 0.95:1;

[0052] Degum, dissolve, dialyze, and dissolve in deionized water the silk to obtain a silk fibroin solution. Then add sodium alginate and stir well until dissolved to obtain Solution B; mix Solution A and Solution B, stir well for 25 min to obtain a mixed crosslinking solution; when preparing Solution A, the reaction mass ratio of oxidized hyaluronic acid, acrylated chitosan, and photoinitiator 2959 is 1:0.8:0.02; when preparing Solution B, the reaction mass ratio of sodium alginate to silk fibroin is 3:3.5; when preparing the mixed crosslinking solution, the reaction mass ratio of oxidized hyaluronic acid to silk fibroin is 1.3:3;

[0053] Step 2: The mixed crosslinking solution is successively subjected to ultrasonic degassing, freezing overnight, and freeze-drying to obtain an aerogel; then the aerogel is impregnated in a calcium chloride solution and photo-crosslinked under ultraviolet light conditions for 5 min, followed by washing with water and freeze-drying to obtain the finished product; the calcium chloride solution is 1.7 wt%; the ultraviolet light intensity is 35 mW / cm 2 。

[0054] Comparative Example 2: Remove solution A, and the rest is the same as in Example 1. The specific steps are as follows: Step 1: Add Pluronic F-127 and 1,3,5-trimethylbenzene to the ethanol aqueous solution in sequence. After ultrasonic dispersion to obtain a white emulsion, magnetic stirring is carried out for 40 min. Then add Tris-HCl solution and dopamine hydrochloride in sequence, adjust the pH to 8.7, and react at 30 °C in the dark for 30 h. After the reaction is completed, solid nanoparticles are collected at 5 °C, and then ultrasonic cleaning, centrifugal collection, washing with deionized water, and freeze-drying are carried out to obtain mesoporous polydopamine nanoparticles; the reaction mass ratio of Pluronic F-127, 1,3,5-trimethylbenzene, Tris-HCl, and dopamine hydrochloride is 37:36:9:6.3;

[0055] Add tannic acid, cinnamaldehyde, and lidocaine to the anhydrous ethanol solution, and stir well for 8 min to obtain a reaction solution; disperse the mesoporous polydopamine nanoparticles into the reaction solution, react at 30 °C in the dark for 30 h, and then carry out centrifugation, washing, and freeze-drying to obtain the composite antibacterial material; the concentration of tannic acid in the reaction solution is 1.9 mg / mL, the concentration of cinnamaldehyde in the reaction solution is 0.6 mg / mL, the concentration of lidocaine in the reaction solution is 1.3 mg / mL, and the concentration of mesoporous polydopamine nanoparticles in the reaction solution is 3.2 mg / mL;

[0056] Step 2: Degum, dissolve, dialyze, and dissolve in deionized water the silk to obtain a silk fibroin solution, and then add sodium alginate and stir well until dissolved to obtain solution B; add the composite antibacterial material to solution B and stir well for 25 min to obtain a mixed crosslinking solution; when preparing solution B, the reaction mass ratio of sodium alginate to silk fibroin is 3:3.5; when preparing the mixed crosslinking solution, the reaction mass ratio of silk fibroin to the composite antibacterial material is 3:0.4;

[0057] Step 3: The mixed crosslinking solution is successively subjected to ultrasonic degassing, freezing overnight, and freeze-drying to obtain an aerogel; then the aerogel is impregnated in a calcium chloride solution and photo-crosslinked under ultraviolet light conditions for 5 min, followed by washing with water and freeze-drying to obtain the finished product; the calcium chloride solution is 1.7 wt%; the ultraviolet light intensity is 35 mW / cm 2 。

[0058] Comparative Example 3: Remove Solution B, and the rest is the same as in Example 1. The specific steps are as follows: Step 1: Add Pluronic F-127 and 1,3,5-trimethylbenzene to the ethanol aqueous solution in sequence. After ultrasonic dispersion to obtain a white emulsion, stir magnetically for 40 min, then add Tris-HCl solution and dopamine hydrochloride in sequence, adjust the pH to 8.7, react at 30 °C in the dark for 30 h. After the reaction, collect the solid nanoparticles at 5 °C, and then through ultrasonic cleaning, centrifugal collection, washing with deionized water, and freeze-drying, mesoporous polydopamine nanoparticles are obtained; the reaction mass ratio of Pluronic F-127, 1,3,5-trimethylbenzene, Tris-HCl, and dopamine hydrochloride is 37:36:9:6.3;

[0059] Add tannic acid, cinnamaldehyde, and lidocaine to the anhydrous ethanol solution, stir well for 8 min to obtain a reaction solution; disperse the mesoporous polydopamine nanoparticles into the reaction solution, react at 30 °C in the dark for 30 h, and then through centrifugation, washing, and freeze-drying, a composite antibacterial material is obtained; the concentration of tannic acid in the reaction solution is 1.9 mg / mL, the concentration of cinnamaldehyde in the reaction solution is 0.6 mg / mL, the concentration of lidocaine in the reaction solution is 1.3 mg / mL, and the concentration of mesoporous polydopamine nanoparticles in the reaction solution is 3.2 mg / mL;

[0060] Step 2: Dissolve chitosan in a 2.3 wt% acetic acid solution, stir at 30 °C for 12 h, then add methacrylic anhydride and react in a water bath at 65 °C for 8 h. After the reaction, adjust the pH to neutral, dialyze, and freeze-dry to obtain acrylated chitosan; the mass-volume ratio of chitosan, acetic acid solution, and methacrylic anhydride is 1 g:50 mL:2.3 mL;

[0061] Dissolve hyaluronic acid in deionized water, then add sodium periodate, react at 30 °C in the dark for 30 h. After the reaction, dialyze and freeze-dry to obtain oxidized hyaluronic acid; dissolve the oxidized hyaluronic acid in deionized water to obtain an oxidized hyaluronic acid solution; add acrylated chitosan and photoinitiator 2959 to the ethanol aqueous solution, stir well for 20 min to obtain a chitosan solution; then mix the oxidized hyaluronic acid solution and the chitosan solution, stir well for 20 min to obtain Solution A; when preparing oxidized hyaluronic acid, the reaction mass ratio of hyaluronic acid and sodium periodate is 0.95:1;

[0062] Add the composite antibacterial material to Solution A, stir well for 25 min to obtain a mixed cross-linking solution; when preparing Solution A, the reaction mass ratio of oxidized hyaluronic acid, acrylated chitosan, and photoinitiator 2959 is 1:0.8:0.02; when preparing the mixed cross-linking solution, the reaction mass ratio of oxidized hyaluronic acid and the composite antibacterial material is 1.3:0.4;

[0063] Step 3: Subject the mixed cross-linking solution to ultrasonic degassing, freeze overnight, and then freeze-dry to obtain an aerogel; then immerse the aerogel in a calcium chloride solution and perform photocrosslinking under ultraviolet light conditions for 5 min, followed by washing with water and freeze-drying to obtain the finished product; the calcium chloride solution is 1.7 wt%; the ultraviolet light intensity is 35 mW / cm 2 .

[0064] Detection test:

[0065] Antibacterial performance test: Cut the finished gel prepared by the present invention into specimens of 10×10 mm, and the specimens need to be disinfected in advance. Inoculate the strain in a liquid medium and incubate at 30 °C for 24 h. Prepare a bacterial solution of 1×10 9 CFU / mL by the ten-fold dilution method, and evenly coat the bacterial solution on an agar plate. Then evenly spread the specimen on the plate and incubate with the agar plate containing the evenly distributed bacterial solution at 30 °C for 24 h. Then obtain the number of colonies through a colony counter and calculate the antibacterial rate.

[0066] Mechanical property test: Referring to the standard of GB / T 1040-2006, cut the finished gel prepared by the present invention into specimens of 50×10 mm. The tensile mechanical properties of the specimens in the dry state were measured using an electronic tensile testing machine WDW-0.05. The test parameters were: the tensile speed was 8 mm / min, the clamping distance was 20 mm, and then substitute into the formula to calculate the breaking strength. The results are shown in the following table:

[0067] Antibacterial rate / % <![CDATA[Breaking strength / MPa > Example 1 93.5 0.81 Example 2 93.2 0.80 Example 3 93.0 0.79 Comparative Example 1 71.2 0.77 Comparative Example 2 83.1 0.69 Comparative Example 3 91.7 0.53

[0068] Conclusion: The dosages of Examples 1-3 remain unchanged, and only some reaction parameters are modified. From the experimental data, it can be seen that there are no obvious fluctuations in the various properties of the specimens.

[0069] Comparative Example 1: Remove the composite antibacterial material, and the rest is the same as in Example 1. From the experimental data, it can be seen that compared with Example 1, the antibacterial rate is reduced to 71.2%. The reason for the analysis is that the mesoporous polydopamine nanoparticles, tannic acid, and cinnamaldehyde in the composite antibacterial material all have certain antibacterial activities. Therefore, after removing them, the antibacterial rate decreases.

[0070] Comparative Example 2: Remove Solution A, and the rest is the same as in Example 1. From the experimental data, it can be seen that compared with Example 1, the antibacterial rate is reduced to 83.1%, and the breaking strength is reduced to 0.69 MPa. The reason for the analysis is that Solution A contains a chitosan antibacterial structure and a Schiff base antibacterial structure, so it has good antibacterial activity; in addition, multiple cross-linking networks in Solution A penetrate and cooperate with each other, making the specimen have a certain tensile resistance; therefore, after removing Solution A, the antibacterial rate and the breaking strength decrease.

[0071] Comparative Example 3: Solution B was removed, and the rest was the same as in Example 1. From the experimental data, it can be seen that compared with Example 1, the breaking strength decreased to 0.53 MPa. The reason is that the network cross-linked by silk fibroin and sodium alginate can provide excellent strength, is not easy to deform and shift, and can maintain the filling effect for a long time, enabling the gel to have the ability to resist compression, tension and tearing.

[0072] It should be noted that in this article, relational terms such as first and second are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the term "including", "comprising" or any other variant thereof is intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements not only includes those elements, but also includes other elements not expressly listed, or further includes elements inherent to such process, method, article or device.

[0073] Finally, it should be noted that the above are only the preferred embodiments of the present invention and are not used to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements for some of the technical features. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. Preparation process of fibroin hyaluronic acid double-network gel for nasolabial fold filling, characterized in that: It includes the following steps: Step 1: Add tannic acid, cinnamaldehyde, and lidocaine to an anhydrous ethanol solution, stir well for 5 - 8 min to obtain a reaction solution; disperse mesoporous polydopamine nanoparticles into the reaction solution, react in the dark at 25 - 30 °C for 25 - 30 h, then centrifuge, wash, and freeze-dry to obtain a composite antibacterial material; Step 2: Dissolve hyaluronic acid in deionized water, then add sodium periodate, react in the dark at 25 - 30 °C for 25 - 30 h, after the reaction, dialyze and freeze-dry to obtain oxidized hyaluronic acid; dissolve the oxidized hyaluronic acid in deionized water to obtain an oxidized hyaluronic acid solution; add acrylated chitosan and photoinitiator 2959 to an ethanol aqueous solution, stir well for 10 - 20 min to obtain a chitosan solution; then mix the oxidized hyaluronic acid solution and the chitosan solution, stir well for 10 - 20 min to obtain solution A; Dissolve silk in degumming, dissolution, dialysis, and deionized water to obtain a silk fibroin solution, then add sodium alginate, stir well until dissolved to obtain solution B; mix solution A and solution B, stir well for 3 - 5 min, then add the composite antibacterial material, continue to stir for 15 - 20 min to obtain a mixed cross-linking solution; Step 3: Subject the mixed cross-linking solution to ultrasonic degassing, freeze overnight, and freeze-dry to obtain an aerogel; then immerse the aerogel in a calcium chloride solution, perform photocrosslinking under ultraviolet light conditions for 3 - 5 min, then wash with water and freeze-dry to obtain the finished product.

2. The preparation process of the fibroin hyaluronic acid double-network gel for nasolabial fold filling according to claim 1, characterized in that: In step 1, the concentration of tannic acid in the reaction solution is 1.8 - 2.0 mg / mL, the concentration of cinnamaldehyde in the reaction solution is 0.5 - 0.7 mg / mL, the concentration of lidocaine in the reaction solution is 1.0 - 1.5 mg / mL, and the concentration of mesoporous polydopamine nanoparticles in the reaction solution is 3.0 - 3.5 mg / mL.

3. The preparation process of the silk fibroin hyaluronic acid double-network gel for nasolabial fold filling according to claim 1, characterized in that: In step 1, the preparation process of mesoporous polydopamine nanoparticles is as follows: Add Pluronic F-127 and 1,3,5-trimethylbenzene to an ethanol aqueous solution in sequence, ultrasonically disperse to obtain a white emulsion, then stir magnetically for 30 - 40 min, then add Tris-HCl solution and dopamine hydrochloride in sequence, adjust the pH to 8.5 - 8.7, react in the dark at 25 - 30 °C for 25 - 30 h, after the reaction, collect solid nanoparticles at 3 - 5 °C, then ultrasonically clean, centrifuge and collect, wash with deionized water, and freeze-dry to obtain mesoporous polydopamine nanoparticles.

4. The preparation process of the silk fibroin - hyaluronic acid double - network gel for nasolabial fold filling according to claim 3, characterized in that: The reaction mass ratio of Pluronic F-127, 1,3,5-trimethylbenzene, Tris-HCl, and dopamine hydrochloride is 37:36:9:(6.0 - 6.5).

5. The preparation process of the silk fibroin - hyaluronic acid double - network gel for nasolabial fold filling according to claim 1, characterized in that: In step 2, the preparation process of acrylated chitosan is as follows: Dissolve chitosan in a 2.0 - 2.5 wt% acetic acid solution, stir at 25 - 30 °C for 10 - 12 h, then dropwise add methacrylic anhydride and react in a water bath at 60 - 65 °C for 6 - 8 h, after the reaction, adjust the pH to neutral, dialyze, and freeze-dry to obtain acrylated chitosan.

6. The preparation process of the fibroin hyaluronic acid double-network gel for nasolabial fold filling according to claim 5, characterized in that: The mass-volume ratio of chitosan, acetic acid solution, and methacrylic anhydride is 1 g : 50 mL : (2.0 - 2.5) mL.

7. The preparation process of the fibroin hyaluronic acid double-network gel for nasolabial fold filling according to claim 1, characterized in that: In step two, when preparing oxidized hyaluronic acid, the reaction mass ratio of hyaluronic acid to sodium periodate is (0.9 - 1.0) :

1.

8. The preparation process of the fibroin hyaluronic acid double-network gel for nasolabial fold filling according to claim 1, characterized in that: In step two, when preparing solution A, the reaction mass ratio of oxidized hyaluronic acid, acrylated chitosan, and photoinitiator 2959 is 1 : (0.7 - 0.9) : 0.02; when preparing solution B, the reaction mass ratio of sodium alginate to silk fibroin is 3 : (3 - 4); when preparing the mixed cross-linking solution, the reaction mass ratio of oxidized hyaluronic acid, silk fibroin, and the composite antibacterial material is (1.2 - 1.5) : 3 : (0.3 - 0.5).

9. The preparation process of the silk fibroin - hyaluronic acid double - network gel for nasolabial fold filling according to claim 1, characterized in that: In step 3, the calcium chloride solution is 1.5 - 2.0 wt%; the ultraviolet light intensity is 25 - 35 mW / cm 2 .

10. A silk fibroin - hyaluronic acid double - network gel for nasolabial fold filling, characterized in that, Prepared according to the preparation process described in any one of claims 1 - 9.