Tough fish scale glue Janus membrane for promoting wound healing and preparation method and application thereof

Abstract

The invention belongs to the field of medical preparations, and particularly relates to a tough fish scale glue Janus membrane for promoting wound healing as well as a preparation method and application of the tough fish scale glue Janus membrane. In the preparation method of the tough fish scale gelatin Janus membrane, chitosan with an antibacterial function and fish scale gelatin with various biological activities are selected as raw materials, a tough Janus membrane dressing capable of being matched with a frequently-moving active wound surface is prepared through a two-step film casting method, the outer chitosan compact layer is antibacterial, and the outer layer is not prone to damage. The inner fish scale gelatin porous layer adheres to the wound surface and maintains a suitable healing microenvironment for wound surface healing, the synergistic effect of inhibiting bacteria, balancing acute inflammatory response and promoting cell proliferation and migration is achieved, repair and regeneration of infectious activity wound surfaces in tissue engineering can be obviously accelerated, industrial production can be achieved easily, and good clinical application prospects are achieved.

Description

Technical Field

[0001] The invention belongs to the field of medical preparations, and in particular relates to a tough fish scale glue Janus film for promoting wound healing, and a preparation method and application thereof. Background Art

[0002] For wounds in vulnerable active parts of the human body such as elbows, knees, ankles, and neck and back, due to frequent movement and stress concentration effects, traditional dressings are not only easy to fall off, have poor mechanical properties, single functions, and are susceptible to bacterial invasion and infection, but also cause secondary injuries, pain, and even disability. Clinical wounds are often treated with tissue staples, clips, or sutures, but they damage the surrounding healthy skin tissue and increase the risk of scarring, infection, inflammation, and surgical complications.

[0003] In recent years, wound dressings such as hydrogels and asymmetric (Janus) membranes have been developed to address the serious problems of secondary injury, tissue fluid exudation, bacterial infection, dehydration, wound ulceration, and scar formation in the healing process of active wounds. In particular, the Janus membrane, which consists of a dense outer layer and a porous inner layer, has attracted much attention because its structure is similar to that of skin tissue. The dense layer of the Janus membrane can mimic the epidermis of the skin, protect the body from bacterial infection and control gas exchange; while the porous inner layer can absorb exudates, promote cell adhesion and proliferation, keep the wound environment moist, and will not destroy new granulation or epithelial tissue. However, the existing Janus wound dressing strategy mainly focuses on biochemical functions, with complex manufacturing processes and high costs, which seriously hinders its clinical application. In addition, the currently developed Janus membrane wound dressings have poor flexibility and cannot match active wounds with frequent movements, which greatly limits its clinical application in wounds in sports joints and other parts.

[0004] Fish scales are mainly composed of type I collagen (about 45%), hydroxyapatite (about 55%) and trace elements (zinc and magnesium). Fish scale glue, as an organic extract of the intercellular matrix of fish scales, is rich in sources, has good biocompatibility and contains trace elements (zinc and magnesium); in particular, it can restore cell function by enhancing cell signaling and cell communication and enhancing cytokine sensitivity, normalize acute inflammatory reactions in wounds, promote cell proliferation and migration, and thus accelerate wound healing. Chitosan is naturally non-toxic, has excellent biocompatibility, and has hemostatic and antibacterial functions as a wound dressing. At present, fish scale collagen has been used for wound dressings. For example, CN110665053B discloses a medical composite collagen sponge dressing, which contains fish scale collagen, bacterial protein and nutrients of culture medium. CN105107011A discloses a fish scale collagen hemostatic sponge, which is made by cross-linking fish scale collagen with chitosan. However, the fish scale collagen in the above literature is pure collagen, lacking trace elements and biological structure in fish scale glue, and has a high preparation cost. In addition, the dressings are all sponge dressings, which are still traditional dressings and cannot fit the wound surface, especially active wounds caused by frequent exercise.

[0005] Therefore, the technical field is in urgent need of a tough fish scale glue Janus membrane wound dressing for matching infectious active wounds and promoting their healing. The dressing can be adhered to the infectious active wound to maintain a suitable wound healing microenvironment and effectively inhibit bacteria, thereby accelerating the healing of infectious active wounds. Summary of the invention

[0006] In order to solve the above technical problems, the present invention provides a tough fish scale glue Janus membrane for promoting wound healing, especially healing of infectious active wounds, and its preparation method and application. The present invention combines the advantages of two types of biomaterials, fish scale glue and chitosan, and combines chitosan with antibacterial function and fish scale glue with multiple biological activities to make a Janus membrane dressing that can match the wound surface of the active part. The outer chitosan dense layer is antibacterial, and the inner fish scale glue porous layer adheres to the wound surface and maintains a suitable healing microenvironment for wound healing, achieving a synergistic effect of antibacterial, balancing acute inflammatory response and promoting healing.

[0007] The first aspect of the present invention provides a method for preparing a tough fish scale glue Janus film for promoting wound healing, comprising the following steps:

[0008] 1) dispersing fish scale glue in a moisturizer solution, and stirring thoroughly until a uniform and completely clear fish scale glue solution is obtained; centrifuging, pouring the obtained fish scale glue supernatant solution into a plate, and drying to obtain a fish scale glue film;

[0009] 2) dissolving chitosan in the solution to obtain a uniform and transparent chitosan solution; then, evenly pouring the obtained chitosan solution on the fish scale film obtained in step 1), and drying to obtain a chitosan / fish scale film;

[0010] 3) placing the chitosan / fish scale glue film obtained in step 2) in a cross-linking reactor, adding a cross-linking agent for cross-linking, and after demolding, obtaining the tough fish scale glue Janus film.

[0011] In some embodiments, in step 1), fish scale glue is prepared by the following method: fresh fish scales are decellularized using 0.5-5wt% Triton X-100, 1-5wt% sodium dodecyl sulfate, 1-20μg / mL DNase and 1-20μg / mL RNase in sequence; then, the decellularized fish scales are treated with 20-40% quicklime and 5-15% citric acid in sequence, and then ultrasonically treated with ultrasound-assisted technology to heat to 80-95°C, ultrasonicate for 1-3 hours, filter, collect the filtrate and dry it to obtain fish scale glue.

[0012] In some embodiments, the fresh fish scales are selected from one or more of tilapia scales, crucian carp scales, carp scales, grass carp scales, silver carp scales, sea bass scales, bighead carp scales, and crispy snapper scales.

[0013] In some embodiments, in step 1), the moisturizer is selected from one or more of glycerol, propylene glycol, butylene glycol, glyceryl polyether-26, allantoin, betaine and trehalose, and the concentration of the moisturizer solution is 5-25wt%. By adding the moisturizer, the flexibility and porosity of the Janus membrane are effectively increased.

[0014] In some embodiments, in step 1), the thickness of the obtained fish scale film is 0.2-0.8 mm.

[0015] In some embodiments, in step 2), chitosan is dissolved in a 0.01-0.1 M acetic acid solution to obtain a 1-2 wt % chitosan-acetic acid solution.

[0016] In some embodiments, in step 1) and step 2), the drying is ventilation drying in a drying oven, and the ventilation drying conditions are: humidity 25%-50%, temperature 18-35°C, and drying time 36-72 hours.

[0017] In some embodiments, in step 3), the crosslinking method selected for crosslinking is steam or immersion crosslinking; the crosslinking agent used for steam crosslinking is formaldehyde or glutaraldehyde, the temperature is 15-35°C, and the crosslinking time is 6-48 hours; the crosslinking agent used for immersion crosslinking is formaldehyde, glutaraldehyde or genipin, the crosslinking agent concentration is 2%-8%, and the crosslinking time is 2-12 hours.

[0018] In some embodiments, in step 3), the thickness of the obtained tough fish scale glue Janus film is 0.4-1.8 mm.

[0019] A second aspect of the present invention provides a tough fish scale glue Janus film for promoting wound healing, which is prepared by the preparation method of any of the above embodiments.

[0020] The third aspect of the present invention provides the application of the tough fish scale glue Janus film that promotes wound healing as a medical dressing for wound healing in the field of wound repair at the wrist, knee, elbow, ankle and neck back in tissue engineering.

[0021] Compared with the prior art, the beneficial effects of the present invention include at least:

[0022] (1) The fish scale glue and chitosan raw materials used in the preparation of the tough fish scale glue Janus membrane dressing of the present invention are both derived from non-mammals, which can effectively avoid the risks of cross-species viral infection and zoonosis; and the fish scale glue fully combines the effective antibacterial function of chitosan and the function of reversing acute inflammatory response and promoting cell proliferation and migration, thereby synergistically promoting wound healing; different from fish scale collagen, the fish scale glue prepared by the present invention retains trace elements and biological structure, has high air permeability and water absorption, and has low cost.

[0023] (2) The preparation method provided by the present invention prepares a tough fish scale glue Janus membrane with excellent flexibility, which has excellent tensile fatigue resistance and matches the wound surface of the active part, and can effectively avoid the falling off of the Janus membrane wound dressing caused by frequent movement of the wound surface.

[0024] (3) The tough fish scale glue Janus membrane dressing of the present invention is a moist wound dressing with asymmetric double-sided functions. The dense outer layer of chitosan has antibacterial and antibacterial functions, which effectively prevents delayed wound healing caused by microbial infection; the inner layer of the fish scale glue porous structure has good hydrophilicity and adhesion, and provides a suitable moist microenvironment for the healing of wounds in active areas, and has excellent clinical medical value;

[0025] (4) The chitosan and fish scale glue used in the present invention can be derived from shrimp and crab shells and marine fish scales, which also provides an effective strategy for the comprehensive development and high-value utilization of marine aquatic product processing waste, and can effectively reduce environmental pollution and resource waste. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 The scanning electron microscope photographs of the macroscopic morphology and cross-sectional structure of the tough fish scale glue Janus membrane prepared in Examples 1 to 4 of the present invention for promoting healing of infectious active wounds;

[0027] Figure 2 This is a mechanical property analysis diagram of the tough fish scale glue Janus film prepared in Example 2-3 of the present invention;

[0028] Figure 3 This is a diagram showing the effect of the tough fish scale gel Janus membrane prepared in Example 2-3 of the present invention on promoting the migration of L929 cells;

[0029] Figure 4 This is a diagram showing the effect of the tough fish scale glue Janus membrane prepared in Example 2-3 of the present invention on promoting the healing of infectious active wounds. DETAILED DESCRIPTION

[0030] In order to make the purpose, technical scheme and beneficial effects of the present invention clearer, the present invention is further described in detail below in conjunction with the accompanying drawings and specific embodiments. Examples of the embodiments are shown in the accompanying drawings. It should be understood that the specific embodiments described in the following embodiments of the present invention are only exemplary descriptions of the specific embodiments of the present invention, and are intended to be used to explain the present invention, but do not constitute a limitation of the present invention.

[0031] The endpoints of ranges and any values ​​disclosed herein are not limited to the exact range or value, and these ranges or values ​​should be understood to include approximations to these ranges.

[0032] Example 1

[0033] A method for preparing a tough fish scale glue Janus film for promoting healing of infectious active wounds comprises the following steps:

[0034] 1) Take a portion of fresh crispy fish scales and decellularize them using 1% Triton X-100, 0.5% sodium dodecyl sulfate, DNase (5 μg / mL) and RNase (5 μg / mL) in sequence; then, treat the decellularized fish scales with 20% quicklime and 10% citric acid in sequence, and then perform ultrasonic treatment and heating (95°C) using ultrasound-assisted technology for 2 hours, collect the filtrate after filtration and freeze-dry to obtain fish scale glue.

[0035] 2) Disperse 0.8 g of fish scale glue in 8 ml of 5% glycerol solution, and stir thoroughly to obtain a uniform and completely clear fish scale glue solution; after centrifugation at 10,000 rpm / min for 15 minutes, pour the fish scale glue supernatant into a polytetrafluoroethylene plate, and dry it in a drying oven at a humidity of 40% and a temperature of 25° C. for 72 hours to obtain a fish scale glue film with a thickness of 0.3 mm.

[0036] 3) dissolving 0.2 g of chitosan in an acetic acid solution with a concentration of 0.01 M, stirring continuously until completely dissolved, to obtain a uniform and transparent chitosan-acetic acid solution with a mass concentration of 2%; then, evenly pouring the obtained chitosan-acetic acid solution on the fish scale film obtained in step 2), drying in a drying oven at a humidity of 40% and a temperature of 25° C. for 72 hours, to obtain a chitosan / fish scale film with a thickness of 0.7 mm.

[0037] 4) The chitosan / fish scale glue film obtained in step 3) and applied to a polytetrafluoroethylene plate was placed in a formaldehyde vapor atmosphere device and cross-linked at 25° C. for 24 hours. After demolding, the tough fish scale glue Janus film (5FSG / CS) was obtained.

[0038] Example 2

[0039] A method for preparing a tough fish scale glue Janus film for promoting healing of infectious active wounds comprises the following steps:

[0040] 1) A portion of fresh sea bass scales was decellularized using 2% Triton X-100, 2% sodium dodecyl sulfate, DNase (5 μg / mL) and RNase (5 μg / mL) in sequence; then, the decellularized scales were treated with 30% quicklime and 15% citric acid in sequence, and then ultrasonically treated and heated (85°C) using ultrasound-assisted technology for 2 hours, and the filtrate was filtered, collected and freeze-dried to obtain fish scale glue.

[0041] 2) 1.6 g of fish scale glue was dispersed in 8 ml of 20% glycerol solution, and the solution was stirred thoroughly until it was uniform and completely clarified; after centrifugation at 10,000 rpm / min for 15 minutes, the fish scale glue supernatant was poured into a polytetrafluoroethylene plate, and dried in a drying oven at a humidity of 30% and a temperature of 25° C. for 48 hours to obtain a fish scale glue film with a thickness of 0.8 mm.

[0042] 3) dissolving 0.2 g of chitosan in an acetic acid solution with a concentration of 0.01 M, stirring continuously until completely dissolved, to obtain a uniform and transparent chitosan-acetic acid solution with a mass concentration of 2%; then, evenly pouring the obtained chitosan-acetic acid solution on the fish scale film obtained in step 2), drying in a drying oven at a humidity of 30% and a temperature of 30° C. for 48 hours, to obtain a chitosan / fish scale film with a thickness of 1.8 mm.

[0043] 4) The chitosan / fish scale glue film obtained in step 3) and applied to a polytetrafluoroethylene plate was placed in a glutaraldehyde vapor atmosphere device and cross-linked at 30° C. for 24 hours. After demolding, the tough fish scale glue Janus film (20FSG / CS) was obtained.

[0044] Example 3

[0045] A method for preparing a tough fish scale glue Janus film for promoting healing of infectious active wounds comprises the following steps:

[0046] 1) A portion of fresh grass carp scales was decellularized using 5% TritonX-100, 3% sodium dodecyl sulfate, DNase (5 μg / mL) and RNase (5 μg / mL) in sequence; then, the decellularized scales were treated with 40% quicklime and 15% citric acid in sequence, and then ultrasonically treated and heated (90°C) using ultrasound-assisted technology for 3 hours, and the filtrate was filtered, collected and freeze-dried to obtain fish scale glue.

[0047] 2) 1.6 g of fish scale glue was dispersed in 8 ml of 10% glycerol solution, and the solution was stirred thoroughly until it was uniform and completely clarified; after centrifugation at 10000 rpm / min for 15 minutes, the fish scale glue supernatant was poured into a polytetrafluoroethylene plate, and dried in a drying oven at a humidity of 30% and a temperature of 35° C. for 72 hours to obtain a fish scale glue film with a thickness of 0.5 mm.

[0048] 3) dissolving 0.2 g of chitosan in an acetic acid solution with a concentration of 0.01 M, stirring continuously until completely dissolved, to obtain a uniform and transparent chitosan-acetic acid solution with a mass concentration of 2%; then, evenly pouring the obtained chitosan-acetic acid solution on the fish scale film obtained in step 2), drying in a drying oven at a humidity of 30% and a temperature of 35° C. for 72 hours, to obtain a chitosan / fish scale film with a thickness of 1.2 mm.

[0049] 4) The chitosan / fish scale glue film obtained in step 3) and applied to a polytetrafluoroethylene plate was placed in a glutaraldehyde vapor atmosphere device and cross-linked at 25° C. for 48 hours. After demolding, the tough fish scale glue Janus film (10FSG / CS) was obtained.

[0050] Example 4

[0051] A method for preparing a tough fish scale glue Janus film for promoting healing of infectious active wounds comprises the following steps:

[0052] 1) A portion of fresh sea bass scales was decellularized using 4% Triton X-100, 2% sodium dodecyl sulfate, DNase (5 μg / mL) and RNase (5 μg / mL) in sequence; then, the decellularized scales were treated with 40% quicklime and 15% citric acid in sequence, and then ultrasonically treated and heated (95°C) using ultrasound-assisted technology for 2 hours, and the filtrate was filtered, collected and freeze-dried to obtain fish scale glue.

[0053] 2) 1.2 g of fish scale glue was dispersed in 8 ml of 15% glycerol solution, and the solution was stirred thoroughly until it was uniform and completely clarified; after centrifugation at 10,000 rpm / min for 15 minutes, the fish scale glue supernatant was poured into a polytetrafluoroethylene plate, and dried in a drying oven at a humidity of 40% and a temperature of 25° C. for 72 hours to obtain a fish scale glue film with a thickness of 0.4 mm.

[0054] 3) dissolving 0.1 g of chitosan in an acetic acid solution having a concentration of 0.01 M, stirring continuously until completely dissolved, to obtain a uniform and transparent chitosan-acetic acid solution having a mass concentration of 1%; then, evenly pouring the obtained chitosan-acetic acid solution on the fish scale film obtained in step 2), and drying in a drying oven at a humidity of 40% and a temperature of 25° C. for 72 hours to obtain a chitosan / fish scale film having a thickness of 0.9 mm.

[0055] 4) The chitosan / fish scale glue film obtained in step 3) and applied to a polytetrafluoroethylene plate was placed in a formaldehyde vapor atmosphere device and cross-linked at 35° C. for 72 hours. After demolding, the tough fish scale glue Janus film (15FSG / CS) was obtained.

[0056] Example 5

[0057] A method for preparing a tough fish scale glue Janus film for promoting healing of infectious active wounds comprises the following steps:

[0058] 1) A portion of fresh grass carp scales was decellularized using 5% Triton X-100, 1% sodium dodecyl sulfate, DNase (5 μg / mL) and RNase (5 μg / mL) in sequence; then, the decellularized scales were treated with 20% quicklime and 10% citric acid in sequence, and then ultrasonically treated and heated (95°C) using ultrasound-assisted technology for 3 hours, and the filtrate was filtered, collected and freeze-dried to obtain fish scale glue.

[0059] 2) 1.2 g of fish scale glue was dispersed in 8 ml of 20% glycerol solution, and the solution was stirred thoroughly until it was uniform and completely clarified; after centrifugation at 10,000 rpm / min for 15 minutes, the fish scale glue supernatant was poured into a polytetrafluoroethylene plate, and dried in a drying oven at a humidity of 30% and a temperature of 35° C. for 48 hours to obtain a fish scale glue film with a thickness of 0.6 mm.

[0060] 3) dissolving 0.2 g of chitosan in an acetic acid solution with a concentration of 0.01 M, stirring continuously until completely dissolved, to obtain a uniform and transparent chitosan-acetic acid solution with a mass concentration of 2%; then, evenly pouring the obtained chitosan-acetic acid solution on the fish scale film obtained in step 2), drying in a drying oven at a humidity of 30% and a temperature of 35° C. for 48 hours, to obtain a chitosan / fish scale film with a thickness of 1.6 mm.

[0061] 4) The chitosan / fish scale glue film obtained in step 3) and applied to the polytetrafluoroethylene plate is immersed in a container containing a glutaraldehyde solution with a mass concentration of 2.5%, cross-linked at room temperature for 6 hours, and then demolded to obtain the tough fish scale glue Janus film.

[0062] In order to verify the beneficial effect of the present invention in promoting the healing of infectious active wounds, the tough fish scale glue Janus films prepared in Examples 1-4 were subjected to the following tests.

[0063] (1) Morphological characteristics and cross-sectional microstructure characterization of tough fish scale glue Janus film

[0064] This experiment characterizes the morphology and cross-sectional microstructure of the tough fish scale glue Janus film prepared in Examples 1-4. Figure 1 It can be seen that the tough fish scale glue Janus film prepared in Examples 1-4 has excellent film-forming properties and good transparency; further scanning electron microscopy shows that the tough fish scale glue Janus film presents an asymmetric double-layer structure, the outer chitosan layer is dense, and the inner fish scale glue layer presents an interconnected porous structure, and there is no obvious break in the connecting part between the double layers.

[0065] (2) Mechanical properties analysis of tough fish scale glue Janus film

[0066] Figure 2 The tensile strength and tensile fatigue resistance of the tough fish scale glue Janus film prepared in Examples 2 and 4 of the present invention. Figure 2 As can be seen from A and B, the tough fish scale glue Janus film has excellent deformation performance in the tensile test, and its deformation is as high as more than 200%. Figure 2 As can be seen in Figure C, the tough fish scale glue Janus membrane can still maintain the original strain after 50 cycles under 200% deformation test conditions. This shows that the prepared fish scale glue Janus membrane has good mechanical properties, and its tensile fatigue resistance can provide mechanical performance support in the wound area of ​​the active part with frequent movement.

[0067] (3) Evaluation of the effect of tough fish scale gel Janus membrane on promoting L929 cell migration

[0068] Figure 3The results of the evaluation of the effect of the tough fish scale glue Janus membrane prepared in Examples 2 and 4 of the present invention on promoting the migration of L929 cells are shown in the figure, wherein the control group is a cell scratch group without the tough fish scale glue Janus membrane prepared in Examples 2 and 4. The results of the cell scratch experiment on the simulated wound surface in vitro showed that within 24 hours, the migration rate of L929 cells in the tough fish scale glue Janus membrane group exceeded 22%, while that in the control group was only 16%; at the 72nd hour, the migration rate of L929 cells in the tough fish scale glue Janus membrane group was about 91%, while that in the control group was only 58%. This shows that the tough fish scale glue Janus membrane can significantly promote the cell migration of L929 cells, thereby facilitating the rapid healing of the wound surface.

[0069] (4) Evaluation of the effect of tough fish scale glue Janus membrane on promoting healing of infectious active wounds

[0070] Figure 4 The results of the healing effect of the tough fish scale glue Janus membrane prepared in Examples 2 and 4 of the present invention on the infectious active wounds of rats are shown in the figure, wherein the control group is the infectious active wounds of rats not covered with the tough fish scale glue Janus membrane prepared in Examples 2-3. Figure 4 The results show that on the 3rd day, the infectious active wounds of the rats in the control group had severe inflammatory reactions, accompanied by the outflow of purulent tissue exudates, while the wounds of the tough fish scale glue Janus membrane groups prepared in Examples 2 and 4 had basically scabbed; in addition, compared with the control group, the number of Staphylococcus aureus colonies in the wounds of the tough fish scale glue Janus membrane groups prepared in Examples 2-3 was significantly reduced. On the 15th day, the infectious active wounds of the tough fish scale glue Janus membrane groups prepared in Examples 2-3 were basically completely healed, while the wound healing rate of the rats in the control group was only about 84%.

[0071] Through the above embodiments and experiments, it can be seen that the present invention combines chitosan with antibacterial function and fish scale glue with multiple biological activities to obtain a tough fish scale glue Janus film dressing. The tough fish scale glue Janus film dressing can not only match the wound surface of the active part, but also its outer chitosan dense layer is antibacterial, and the inner fish scale glue porous layer adheres to the wound surface and maintains a suitable healing microenvironment for wound healing, achieving the synergistic effect of antibacterial, balancing acute inflammatory response and promoting healing, and can significantly promote the repair and regeneration of infectious active wounds in tissue engineering, and has a good application prospect.

[0072] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing a tough fish scale glue Janus film that promotes wound healing, characterized in that: The steps include: 1) dispersing fish scale glue in a moisturizer solution, stirring thoroughly to obtain a uniform and completely clear fish scale glue solution; centrifuging, pouring the obtained fish scale glue supernatant solution into a plate, and drying to obtain a fish scale glue film; 2) dissolving chitosan in the solution to obtain a uniform and transparent chitosan solution; then, evenly pouring the obtained chitosan solution on the fish scale film obtained in step 1), and drying to obtain a chitosan / fish scale film; 3) placing the chitosan / fish scale glue film obtained in step 2) in a cross-linking reactor, adding a cross-linking agent for cross-linking, and after demolding, obtaining the tough fish scale glue Janus film.

2. The method for preparing the tough fish scale glue Janus film for promoting wound healing as claimed in claim 1, characterized in that: In step 1), fish scale glue is prepared by the following method: fresh fish scales are decellularized using 0.5-5wt% Triton X-100, 1-5wt% sodium dodecyl sulfate, 1-20μg / mL DNase and 1-20μg / mL RNase in sequence; Then, the decellularized fish scales are treated with 20-40% quicklime and 5-15% citric acid in turn, and then ultrasonically treated by heating to 80-95°C for 1-3 hours using ultrasonic-assisted technology. After filtering, the filtrate is collected and dried to obtain fish scale glue.

3. The method for preparing the tough fish scale glue Janus film for promoting wound healing as claimed in claim 2, characterized in that: The fresh fish scales are selected from one or more of tilapia scales, crucian carp scales, carp scales, grass carp scales, silver carp scales, sea bass scales, bighead carp scales, and crispy mullet scales.

4. The method for preparing the tough fish scale glue Janus film for promoting wound healing as claimed in claim 1, characterized in that: In step 1), the moisturizing agent is selected from one or more of glycerol, propylene glycol, butylene glycol, glycerol polyether-26, allantoin, betaine and trehalose, the concentration of the moisturizing agent solution is 5-25wt%, and the thickness of the obtained fish scale film is 0.2-0.8mm.

5. The method for preparing the tough fish scale glue Janus film for promoting wound healing as claimed in claim 1, characterized in that: In step 2), chitosan is dissolved in a 0.01-0.1 M acetic acid solution to obtain a 1-2 wt % chitosan-acetic acid solution.

6. The method for preparing the tough fish scale glue Janus film for promoting wound healing as claimed in claim 1, characterized in that: In step 1) and step 2), the drying is performed by ventilation drying in a drying oven, and the ventilation drying conditions are: humidity 25%-50%, temperature 18-35° C., and drying time 36-72 hours.

7. The method for preparing the tough fish scale glue Janus film for promoting wound healing as claimed in claim 1, characterized in that: In step 3), the crosslinking method selected for crosslinking is steam or immersion crosslinking; the crosslinking agent used for steam crosslinking is formaldehyde or glutaraldehyde, the temperature is 15-35°C, and the crosslinking time is 6-48 hours; the crosslinking agent used for immersion crosslinking is formaldehyde, glutaraldehyde or genipin, the crosslinking agent concentration is 2%-8%, and the crosslinking time is 2-12 hours.

8. The method for preparing the tough fish scale glue Janus film for promoting wound healing as claimed in claim 1, characterized in that: In step 3), the thickness of the obtained tough fish scale glue Janus film is 0.4-1.8 mm.

9. A tough fish scale glue Janus membrane for promoting wound healing, characterized in that: The preparation is obtained by the preparation method according to any one of claims 1 to 8.

10. Use of the tough fish scale glue Janus membrane for promoting wound healing as claimed in claim 9 as a wound dressing in the field of wound repair at the wrist, knee, elbow, ankle and neck back in tissue engineering.

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