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

Abstract

The application relates to the technical field of ophthalmic material preparation, and particularly discloses a silk fibroin composite material and application thereof in preparation of corneal injury repair materials. The silk fibroin composite material contains silk fibroin and mussel mucin; the silk fibroin composite material further contains an active ingredient; the active ingredient is selected from spinozine, erythronol or alderone. Research shows that the silk fibroin composite material prepared by adding spinozine, erythronol or alderone in the preparation process has a significant analgesic effect; therefore, the silk fibroin composite material is used for preparing the corneal injury repair material, and has important application value for relieving pain of a wound after corneal injury surgical repair.

Description

Technical Field

[0001] This invention relates to the field of ophthalmic material preparation technology, specifically to a silk fibroin composite material and its application in the preparation of corneal damage repair materials. Background Technology

[0002] The cornea is a transparent, avascular fibrous membrane located in the anterior 1 / 6 of the eyeball wall. Corneal injury refers to damage to the cornea caused by external forces or internal factors; it includes corneal lacerations, penetrating and ruptured corneas, corneal abrasions, corneal contusions, and loss of deep corneal tissue. Severe corneal damage requires surgical repair. Corneal repair materials are indispensable in the surgical repair of corneal injuries.

[0003] Chinese invention patent CN201910762378.8 discloses a method for preparing a collagen-based cellulose nanocrystal composite film. The method includes the following steps: preparing aqueous dispersions of collagen solution and cellulose nanocrystals separately; then removing air bubbles from the mixed solution at room temperature, pouring it into a petri dish, and air-drying to obtain the collagen-based cellulose nanocrystal composite film. The composite film prepared by this method exhibits good mechanical properties and excellent 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 adhesive protein. The preparation steps of this method are as follows: regenerated silk fibroin and a certain proportion of marine mussel adhesive protein are dissolved in a certain amount of organic solvent, and the mixture is stirred continuously to make it uniform and prepared to a certain concentration, thereby obtaining a mixed solution of silk fibroin and marine mussel adhesive protein. A certain amount of the mixed solution is used to form a membrane or scaffold. The bio-silk fibroin and marine mussel adhesive protein composite nanofiber membrane prepared by this invention has strong mechanical properties.

[0005] Further research by the inventors revealed that while the aforementioned composite film possesses good mechanical properties, post-operative wound repair following corneal injury surgery often results in pain due to the wear and tear of anesthetic drugs. However, the composite film does not provide pain relief. Therefore, developing a composite material with analgesic properties would have significant application value in alleviating post-operative wound pain associated with 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 the preparation of corneal damage repair materials.

[0007] This invention first provides a silk fibroin composite material, which comprises silk fibroin and mussel adhesive protein; and also contains active ingredients;

[0008] The active ingredient is selected from spinosin, osmidrone, or alder.

[0009] The inventors made a surprising discovery during their research: silk fibroin composites prepared by adding spinosin, scutellarin, or alder ketone exhibited significant analgesic effects. In particular, the analgesic effect of the silk fibroin composite prepared with spinosin was significantly higher than that prepared with scutellarin or alder ketone.

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

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

[0012] Preferably, the active ingredient is composed of spinosin and osmectin.

[0013] Preferably, the mass ratio of spinosin to osmidrone in the active ingredients is 1-3:1-3.

[0014] Preferably, the mass ratio of spinosin to osmidrone in the active ingredients is 1:1.

[0015] Further research by the inventors revealed a surprising discovery: silk fibroin composites prepared by simultaneously adding both spinosin and osmidrone exhibited significantly higher analgesic effects than those prepared by adding either spinosin or osmidrone alone. The simultaneous addition of spinosin and osmidrone during the preparation of silk fibroin composites synergistically enhances their analgesic effect.

[0016] The present invention also provides an application of the above-mentioned silk fibroin composite material in the preparation of corneal damage repair materials.

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

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

[0019] (2) Take the silk fibroin composite liquid to make a film, and obtain the silk fibroin composite film, which is the corneal damage repair material.

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

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

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

[0023] Beneficial effects: This invention provides a novel method for preparing silk fibroin composite materials; the silk fibroin composite materials prepared by adding spinosin, scutellarin or alder ketone during the preparation process have significant analgesic effects; therefore, the silk fibroin composite materials described in this invention are used to prepare corneal injury repair materials, which have important application value in relieving wound pain after corneal injury surgery. Detailed Implementation

[0024] The present invention will be further explained below with reference to specific embodiments, but the embodiments do not limit the scope of protection of the present invention.

[0025] Example 1: Preparation of silk fibroin composite material

[0026] The silk fibroin composite material is composed of silk fibroin, mussel adhesive protein and active ingredients in a mass ratio of 100:3:2;

[0027] The active ingredient is Spinosin.

[0028] Example 2: Preparation of silk fibroin composite material

[0029] The silk fibroin composite material is composed of silk fibroin, mussel adhesive protein and active ingredients in a mass ratio of 100:3:2;

[0030] The active ingredient is osmanthus ketone.

[0031] Example 3: Preparation of silk fibroin composite material

[0032] The silk fibroin composite material is composed of silk fibroin, mussel adhesive protein and active ingredients in a mass ratio of 100:3:2;

[0033] The active ingredient is alder ketone.

[0034] Example 4: Preparation of silk fibroin composite material

[0035] The silk fibroin composite material is composed of silk fibroin, mussel adhesive protein and active ingredients in a mass ratio of 100:3:2;

[0036] The active ingredient is composed of spinoxanone and osmanthus ketone in a mass ratio of 1:1.

[0037] Example 5: Preparation of Silk Fibroin Composite Material

[0038] The silk fibroin composite material is composed of silk fibroin, mussel adhesive protein and active ingredients in a mass ratio of 100:3:2;

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

[0040] Example 6: Preparation of Silk Fibroin Composite Material

[0041] The silk fibroin composite material is composed of silk fibroin, mussel adhesive protein and active ingredients in a mass ratio of 100:3:2;

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

[0043] Experiment Example 1: Analgesia Experiment

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

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

[0046] Experimental methods: Eighty-four male KM mice weighing 18–22 g were randomly divided into seven groups of 12 mice each: experimental groups 1–6 and a control group. Before the experiment, 1 mL of the test sample solution was applied to the paws and hind legs of mice in experimental groups 1–6, while the control group was applied with an equal amount of hexafluoroisopropanol. Three hours after the test sample solution was applied, 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 paws. The time required for the mice to lick their paws was the pain threshold. The pain threshold was recorded, and the results are shown in Table 1.

[0047] Table 1. Results of the analgesic experiment

[0048] Sample 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] As can be seen from the experimental results in Table 1, the pain threshold of experimental groups 1-3 was significantly higher than that of the control group. This indicates that the silk fibroin composite materials prepared by adding spinosin, scutellarin or alder ketone during the preparation process have significant analgesic effects.

[0050] As can be seen from the experimental results in Table 1, the pain threshold of experimental group 1 is significantly higher than that of experimental groups 1 and 2. This indicates that 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 adding styraxone or alder.

[0051] As shown in Table 1, the pain threshold of experimental group 4 was significantly higher than that of experimental groups 1-3. This indicates that the analgesic effect of silk fibroin composites prepared by simultaneously adding both spinosin and osmidrone is significantly higher than that prepared by adding either spinosin or osmidrone alone. Therefore, the simultaneous addition of spinosin and osmidrone during the preparation of silk fibroin composites can synergistically enhance the analgesic effect of the prepared composites.

[0052] As can be seen from the experimental results in Table 1, the pain thresholds of experimental groups 5 and 6 were not significantly higher than those of experimental groups 1-3. This indicates that only by adding both sine and alder ketone simultaneously during the preparation of the silk fibroin composite material can the analgesic effect of the prepared silk fibroin composite material be synergistically improved. However, the combination of adding sine, alder ketone, and the other two of alder ketone did not synergistically improve the analgesic effect of the prepared silk fibroin composite material.

[0053] Example 7: Preparation of Corneal Damage Repair Materials

[0054] (1) Take the silk fibroin composite material prepared in Example 1, dissolve it in a solvent to obtain a silk fibroin composite solution; wherein, the ratio of the amount of silk fibroin composite material to the amount of solvent is 1g:15mL; the solvent is hexafluoroisopropanol;

[0055] (2) The silk fibroin composite liquid is used to make a film by electrospinning technology to obtain the silk fibroin composite film, which is the corneal damage repair material.

[0056] Example 8: Preparation of Corneal Damage Repair Material

[0057] (1) Take the silk fibroin composite material prepared in Example 2, dissolve it in a solvent to obtain a silk fibroin composite solution; wherein, the ratio of the amount of silk fibroin composite material to the amount of solvent is 1g:10mL; the solvent is hexafluoroisopropanol;

[0058] (2) The silk fibroin composite liquid is used to make a film by electrospinning technology to obtain the silk fibroin composite film, which is the corneal damage repair material.

[0059] Example 9: Preparation of Corneal Damage Repair Materials

[0060] (1) Take the silk fibroin composite material prepared in Example 3, dissolve it in a solvent to obtain a silk fibroin composite solution; wherein, the ratio of the amount of silk fibroin composite material to the amount of solvent is 1g:20mL; the solvent is hexafluoroisopropanol;

[0061] (2) The silk fibroin composite liquid is used to make a film by electrospinning technology to obtain the silk fibroin composite film, which is the corneal damage repair material.

[0062] Example 10: Preparation of Corneal Damage Repair Materials

[0063] (1) Take the silk fibroin composite material prepared in Example 4, dissolve it in a solvent to obtain a silk fibroin composite solution; wherein, the ratio of the amount of silk fibroin composite material to the amount of solvent is 1g:15mL; the solvent is hexafluoroisopropanol;

[0064] (2) The silk fibroin composite liquid is used to make a film by electrospinning technology to obtain the silk fibroin composite film, which is the corneal damage repair material.

Claims

1. A silk fibroin composite material comprising silk fibroin and mussel adhesive protein; characterized in that, It also contains active ingredients; The active ingredients consist of spinosin and osmanthus ketone; The mass ratio of silk fibroin, mussel adhesive protein, and active ingredients in the silk fibroin composite material is 100:3:

2. The mass ratio of spinosin to osmidrone in the active ingredients is 1:

1.

2. The application of the silk fibroin composite material according to claim 1 in the preparation of corneal damage repair materials.

3. The application according to claim 2, characterized in that, The corneal damage repair material is prepared by the following method: (1) Take the silk fibroin composite material according to claim 1, dissolve it in a solvent to obtain a silk fibroin composite solution; (2) Take the silk fibroin composite liquid to make a film, and obtain the silk fibroin composite film, which is the corneal damage repair material.

4. The application according to claim 3, characterized in that, The solvent mentioned in step (1) is hexafluoroisopropanol.

5. The application according to claim 3, characterized in that, In step (1), the ratio of silk fibroin composite material to solvent is 1g:10~20mL.

6. The application according to claim 5, characterized in that, In step (1), the ratio of silk fibroin composite material to solvent is 1g:15mL.