Silk fibroin composite material and application thereof in preparation of corneal injury repair material

Abstract

The invention relates to the technical field of ophthalmic material preparation, and particularly discloses a silk fibroin composite material and application thereof in preparation of a corneal injury repair material. The silk fibroin composite material comprises silk fibroin and mussel mucin, the composition further comprises active ingredients; the active component is selected from spinosin, osmundone or alnudinone. Researches show that in the preparation process of the silk fibroin composite material, spinosin, osmundone or alnone is added to prepare the silk fibroin composite material, and the silk fibroin composite material has a remarkable pain relieving effect; therefore, when the silk fibroin composite material is used for preparing a corneal injury repair material, the silk fibroin composite material has important application value for relieving wound pain after corneal injury operation repair.

Description

Technical Field

[0001] The invention relates to the technical field of ophthalmic material preparation, and in particular to a silk fibroin composite material and application thereof in preparing corneal damage repairing material. Background Art

[0002] The cornea is a transparent fiber membrane without blood vessels located in the front 1 / 6 of the eyeball wall. Corneal injury refers to the damage to the cornea caused by external factors or body factors, including corneal interlayer tear, corneal penetration and rupture, corneal abrasion, corneal contusion, deep corneal tissue loss, etc. If the cornea is seriously injured, surgical repair is required. Corneal injury repair materials are indispensable materials in the process of corneal injury surgical repair.

[0003] Chinese invention patent CN201910762378.8 discloses a method for preparing a collagen-based cellulose nanocrystal composite membrane; the preparation method comprises the following steps: preparing a collagen solution and an aqueous dispersion of cellulose nanocrystals respectively, then removing bubbles from the mixed solution at room temperature, pouring it into a culture dish, and air-drying it to obtain a collagen-based cellulose nanocrystal composite membrane. The composite membrane prepared by this method has good mechanical properties and good biocompatibility, and can be used as an alternative material for corneal repair.

[0004] Chinese invention patent CN201310562721.7 discloses a method for preparing a composite material of silk fibroin and marine mussel adhesion protein; the preparation steps of the method are as follows: dissolving regenerated silk fibroin and a certain proportion of marine mussel adhesion protein in a certain amount of organic solvent, stirring continuously to make them evenly mixed, and preparing a certain concentration to obtain a mixed solution of silk fibroin and marine mussel adhesion protein, and a certain amount of the mixed solution is used to make a membrane or a scaffold. The biological silk fibroin and marine mussel adhesion protein composite nanofiber membrane prepared by the invention has strong mechanical properties.

[0005] The inventors found in further research that although the composite film has good mechanical properties, the wound needs to be repaired after corneal injury surgery, and the pain of the wound will be caused after the anesthetic drugs lose their effectiveness. However, the composite film does not have an analgesic effect. Therefore, the development of a composite material with an analgesic effect has important application value for relieving the pain of the wound after corneal injury surgery. Summary of the invention

[0006] In order to solve at least one of the technical problems existing in the prior art, the present invention provides a silk fibroin composite material and its application in preparing corneal damage repair material.

[0007] The present invention first provides a silk fibroin composite material, which comprises silk fibroin and mussel viscose; and also comprises an active ingredient;

[0008] The active ingredient is selected from spinosin, ospermone or alnus ketone.

[0009] The inventors surprisingly found in the research that in the preparation process of the silk fibroin composite material, the silk fibroin composite material prepared by adding spinosin, osfera ketone or alnus ketone has a significant analgesic effect. Among them, the analgesic effect of the silk fibroin composite material prepared by adding spinosin is significantly higher than that of the silk fibroin composite material prepared by osfera ketone or alnus ketone.

[0010] Preferably, the mass ratio of silk fibroin, mussel mucin and active ingredient in the silk fibroin composite material is 100:1-5:1-5.

[0011] Most preferably, the mass ratio of silk fibroin, mussel mucin and active ingredient in the silk fibroin composite material is 100:3:2.

[0012] Preferably, the active ingredients consist of spinosin and porphyrin.

[0013] Preferably, the mass ratio of spinosum to porphyrin in the active ingredient is 1-3:1-3.

[0014] Preferably, the mass ratio of spinosum to porphyrin in the active ingredient is 1:1.

[0015] The inventors surprisingly found in further research that in the preparation process of the silk fibroin composite material, the analgesic effect of the silk fibroin composite material prepared by adding spinotoxin and porphyrin at the same time is much higher than that of the silk fibroin composite material prepared by adding spinotoxin or porphyrin alone. In the preparation process of the silk fibroin composite material, adding spinotoxin and porphyrin at the same time can synergistically improve the analgesic effect of the prepared silk fibroin composite material.

[0016] The present invention also provides an application of the silk fibroin composite material in preparing a corneal damage repair material.

[0017] Preferably, the corneal damage repair material is prepared by the following method:

[0018] (1) taking the silk fibroin composite material according to any one of claims 1 to 4 and dissolving it in a solvent to obtain a silk fibroin composite liquid;

[0019] (2) taking the silk fibroin composite liquid to form a film to obtain a silk fibroin composite film, that is, the corneal damage repair material.

[0020] Preferably, the solvent in step (1) is hexafluoroisopropanol.

[0021] Preferably, in step (1), the ratio of the silk fibroin composite material to the solvent is 1 g: 10-20 mL.

[0022] Preferably, in step (1), the amount ratio of the silk fibroin composite material to the solvent is 1 g:15 mL.

[0023] Beneficial effects: The present invention provides a novel method for preparing a silk fibroin composite material; during the preparation of the silk fibroin composite material, the silk fibroin composite material prepared by adding spinotoxin, osfera ketone or alnus ketone has a significant analgesic effect; therefore, the silk fibroin composite material of the present invention is used to prepare a corneal injury repair material, which has important application value for alleviating wound pain after corneal injury surgical repair. DETAILED DESCRIPTION

[0024] The present invention is further explained below in conjunction with specific examples, but the examples do not limit the protection scope of the present invention.

[0025] Example 1 Preparation of silk fibroin composite material

[0026] The silk fibroin composite material is formed by mixing silk fibroin, mussel viscose and active ingredients in a mass ratio of 100:3:2;

[0027] Wherein, the active ingredient is spinosum.

[0028] Example 2 Preparation of silk fibroin composite material

[0029] The silk fibroin composite material is formed by mixing silk fibroin, mussel viscose and active ingredients in a mass ratio of 100:3:2;

[0030] Wherein, the active ingredient is purpurogenol.

[0031] Example 3 Preparation of silk fibroin composite material

[0032] The silk fibroin composite material is formed by mixing silk fibroin, mussel viscose and active ingredients in a mass ratio of 100:3:2;

[0033] Wherein, the active ingredient is alnus ketone.

[0034] Example 4 Preparation of Silk Fibroin Composite Material

[0035] The silk fibroin composite material is formed by mixing silk fibroin, mussel viscose and active ingredients in a mass ratio of 100:3:2;

[0036] The active ingredient is composed of spinosum and porphyrin in a mass ratio of 1:1.

[0037] Example 5 Preparation of silk fibroin composite material

[0038] The silk fibroin composite material is formed by mixing silk fibroin, mussel viscose and active ingredients in a mass ratio of 100:3:2;

[0039] The active ingredient is composed of spinosadone and alnus ketone in a mass ratio of 1:1.

[0040] Example 6 Preparation of Silk Fibroin Composite Material

[0041] The silk fibroin composite material is formed by mixing silk fibroin, mussel viscose and active ingredients in a mass ratio of 100:3:2;

[0042] The active ingredient is composed of porphyrone and alnus ketone in a mass ratio of 1:1.

[0043] Experimental Example 1 Analgesia Experiment

[0044] The samples to be tested in this experiment are the silk fibroin composite materials prepared in Examples 1 to 6;

[0045] The preparation method of the sample solution to be tested is as follows: the silk fibroin composite material prepared in Examples 1 to 6 is dissolved in hexafluoroisopropanol to obtain the sample solution to be tested; wherein the amount ratio of the silk fibroin composite material to the solvent is 1 g:15 mL.

[0046] Experimental method: 84 male KM mice weighing 18-22g were selected and randomly divided into 7 groups, 12 mice in each group; they were experimental groups 1-6 and a control group; before the experiment, 1 mL of the sample solution to be tested was applied to the soles and hind legs of the mice in the experimental groups 1-6, and the control group was applied with an equal amount of hexafluoroisopropanol; 3 hours after applying the sample solution to be tested, the mice in each group were placed on a hot plate at 50°C to test the time required for the mice to lick their feet. The time required for the mice to lick their feet is the pain threshold, and the pain threshold is recorded. The results are shown in Table 1.

[0047] Table 1. Analgesia test results

[0048] Samples to be tested Pain threshold Control group - 17.9s±1.4s Experimental Group 1 Silk fibroin composite material prepared in Example 1 25.7s±2.6s Experimental Group 2 Silk fibroin composite material prepared in Example 2 22.3s±2.0s Experimental Group 3 Silk fibroin composite material prepared in Example 3 21.5s±2.4s Experimental Group 4 Silk fibroin composite material prepared in Example 4 33.1s±3.5s Experimental Group 5 Silk fibroin composite material prepared in Example 5 23.3s±2.1s Experimental Group 6 Silk fibroin composite material prepared in Example 6 21.9s±2.3s

[0049] From the experimental results in Table 1, it can be seen that the pain threshold of experimental groups 1 to 3 is significantly higher than that of the control group; this shows that in the preparation process of silk fibroin composite materials, the silk fibroin composite materials prepared by adding spinosone, osfera ketone or alnus ketone have significant analgesic effect.

[0050] From the experimental results in Table 1, it can be seen that the pain threshold of experimental group 1 is also significantly higher than that of experimental groups 1 and 2; this shows that: in the preparation process of silk fibroin composite materials, the silk fibroin composite materials prepared by adding spinosin have a significantly higher analgesic effect than the silk fibroin composite materials prepared by adding osfera ketone or alnus ketone.

[0051] From the experimental results in Table 1, it can be seen that the pain threshold of experimental group 4 is much higher than that of experimental groups 1 to 3; this shows that in the preparation process of the silk fibroin composite material, the silk fibroin composite material prepared by adding spinotoxin and osfera ketone at the same time has a much higher analgesic effect than the silk fibroin composite material prepared by adding spinotoxin or osfera ketone alone. In the preparation process of the silk fibroin composite material, adding spinotoxin and osfera ketone at the same time can synergistically improve the analgesic effect of the prepared silk fibroin composite material.

[0052] It can be seen from the experimental results in Table 1 that the pain thresholds of experimental groups 5 and 6 cannot be further significantly higher than those of experimental groups 1 to 3; this shows that in the preparation process of silk fibroin composite materials, only by adding spinotoxin and osfera ketone at the same time can the analgesic effect of the prepared silk fibroin composite materials be synergistically improved; however, adding a combination of spinotoxin, osfera ketone and the other two of alnus ketone cannot synergistically improve the analgesic effect of the prepared silk fibroin composite materials.

[0053] Example 7 Preparation of corneal damage repair material

[0054] (1) The silk fibroin composite material prepared in Example 1 was dissolved in a solvent to obtain a silk fibroin composite liquid; wherein the amount ratio of the silk fibroin composite material to the solvent was 1 g:15 mL; and the solvent was hexafluoroisopropanol;

[0055] (2) Taking the silk fibroin composite liquid and using electrostatic spinning technology to form a film to obtain a silk fibroin composite film, that is, the corneal damage repair material.

[0056] Example 8 Preparation of corneal damage repair material

[0057] (1) The silk fibroin composite material prepared in Example 2 was dissolved in a solvent to obtain a silk fibroin composite liquid; wherein the amount ratio of the silk fibroin composite material to the solvent was 1 g:10 mL; and the solvent was hexafluoroisopropanol;

[0058] (2) Taking the silk fibroin composite liquid and using electrostatic spinning technology to form a film to obtain a silk fibroin composite film, that is, the corneal damage repair material.

[0059] Example 9 Preparation of corneal damage repair material

[0060] (1) The silk fibroin composite material prepared in Example 3 was dissolved in a solvent to obtain a silk fibroin composite liquid; wherein the amount ratio of the silk fibroin composite material to the solvent was 1 g:20 mL; and the solvent was hexafluoroisopropanol;

[0061] (2) Taking the silk fibroin composite liquid and using electrostatic spinning technology to form a film to obtain a silk fibroin composite film, that is, the corneal damage repair material.

[0062] Example 10 Preparation of corneal damage repair material

[0063] (1) The silk fibroin composite material prepared in Example 4 was dissolved in a solvent to obtain a silk fibroin composite liquid; wherein the amount ratio of the silk fibroin composite material to the solvent was 1 g:15 mL; and the solvent was hexafluoroisopropanol;

[0064] (2) Taking the silk fibroin composite liquid and using electrostatic spinning technology to form a film to obtain a silk fibroin composite film, that is, the corneal damage repair material.

Claims

1. A silk fibroin composite material comprising silk fibroin and mussel viscose; characterized in that: It also contains active ingredients; The active ingredient is selected from spinosin, ospermone or alnus ketone.

2. The silk fibroin composite material according to claim 1, characterized in that The mass ratio of silk fibroin, mussel mucin and active ingredients in the silk fibroin composite material is 100:1-5:1-5. Most preferably, the mass ratio of silk fibroin, mussel mucin and active ingredient in the silk fibroin composite material is 100:3:

2.

3. The silk fibroin composite material according to claim 1, characterized in that The active ingredients are composed of spinosin and porphyrone.

4. The silk fibroin composite material according to claim 3, characterized in that: The mass ratio of spinosin to porphyrin in the active ingredients is 1-3:1-3.

5. The silk fibroin composite material according to claim 4, characterized in that: The mass ratio of spinosum to porphyrin in the active ingredients is 1:

1.

6. Use of the silk fibroin composite material according to any one of claims 1 to 5 in preparing corneal damage repair materials.

7. The use according to claim 6, characterized in that: The corneal damage repair material is prepared by the following method: (1) taking the silk fibroin composite material according to any one of claims 1 to 4 and dissolving it in a solvent to obtain a silk fibroin composite liquid; (2) taking the silk fibroin composite liquid to form a film to obtain a silk fibroin composite film, that is, the corneal damage repair material.

8. The use according to claim 7, characterized in that: The solvent described in step (1) is hexafluoroisopropanol.

9. The use according to claim 7, characterized in that: In step (1), the amount ratio of the silk fibroin composite material to the solvent is 1 g: 10-20 mL.

10. The use according to claim 9, characterized in that: In step (1), the amount ratio of the silk fibroin composite material to the solvent is 1 g:15 mL.