A method for preparing a transparent collagen membrane
By repeatedly washing and transparentizing the collagen membrane, a transparent collagen membrane was prepared, which solved the problems of insufficient mechanical properties and aesthetics of existing materials in the repair of corneal dermoid tumors, and achieved good healing effect and aesthetics.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- NEO MODULUS (SUZHOU) MEDICAL SCI TECH CO LTD
- Filing Date
- 2023-08-01
- Publication Date
- 2026-05-29
AI Technical Summary
Existing patch materials have problems such as poor mechanical properties, rapid degradation, and large color difference with surrounding tissues after repairing corneal dermoid tumors, which affect aesthetics.
Collagen membranes were prepared using a transparency treatment method, which included multiple washing, freeze-drying, and transparency treatment steps to maintain the triple helix structure of the collagen membrane and ensure its transparency and mechanical properties.
The prepared transparent collagen membrane has good mechanical and degradation properties, can maintain the original tissue color and shape at the wound site, and promotes aesthetics during the healing process.
Smart Images
Figure CN116850345B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of biomedical materials, and in particular to a method for preparing a transparent collagen membrane. Background Technology
[0002] Corneal dermoid tumors are the most common benign eye tumors. They are congenital abnormalities resembling tumors, but histologically they are not true tumors, but rather typical keratomas. Corneal dermoid tumors are usually present at birth, slightly increasing in size with age and eye development, accounting for approximately 20% of childhood external eye tumors. Corneal dermoid tumors commonly occur in the infratemporal and temporal regions of the limbus, but can also occur anywhere on the cornea. The limbus is often the center of the dermoid tumor, with one half on the cornea and the other half on the scleral surface. A dermoid tumor is a round, flat, yellow or pink, hill-like mass with visible hair on its surface. Corneal dermoid tumors not only affect appearance, but with age, they can also invade the cornea in the pupillary area, affecting vision and causing corneal astigmatism. As astigmatism gradually increases, vision will decrease, leading to amblyopia.
[0003] Since corneal dermoid tumors are congenital benign corneal tumors, drug treatment is ineffective. If the tumor is small and does not affect visual function or appearance, it can be observed temporarily. If the tumor grows and affects vision or appearance, and the mass is relatively superficial, it should be surgically removed as soon as possible. However, if the wound is not repaired after removal, scarring will remain. Most existing patch materials are decellularized matrix materials, such as dermis and small intestinal mucosa; or synthetic polymer materials, such as PGA, PLGA, PCL, hyaluronic acid, etc. These patch materials appear milky white or slightly yellow after synthesis. If used to repair wound areas, they will cause a significant difference in color between the wound area and the normal cornea and sclera, affecting aesthetics. Each person's scleral color also has slight differences. Custom-made patches for aesthetic purposes are expensive, and the color is difficult to control in the long term. At the same time, the helical structure of artificially synthesized collagen is poor, with some helical structures being incomplete, resulting in poor mechanical properties and excessively rapid degradation.
[0004] Therefore, there is an urgent need in this field for a patch material with good mechanical properties, degradation properties, and the ability to maintain the aesthetic appearance of the healed tissue. Summary of the Invention
[0005] The problem the invention aims to solve
[0006] Based on the aforementioned problems in the existing technology, the purpose of this invention is to provide a transparent collagen membrane with good mechanical properties and degradation properties, and which can reflect the original tissue color and morphology of the wound site.
[0007] Solution for solving the problem
[0008] This invention provides a method for preparing a transparent collagen membrane, wherein the method includes the following steps:
[0009] (a) Obtaining decellularized collagen membranes of animal origin;
[0010] (b) Transparent the collagen membrane at least three times.
[0011] Preferably, step (a) includes:
[0012] (a1) Take animal tissue and remove the surface fat and impurities;
[0013] (a2) Processing animal tissues after removing surface fat and impurities;
[0014] (a3) The animal tissue treated in step (a2) was freeze-dried using a freeze dryer to obtain a dry decellularized collagen membrane.
[0015] Preferably, the treatment includes: washing with a sodium chloride solution;
[0016] More preferably, the treatment includes: washing with sodium chloride solution, treating with hydrochloric acid, and then washing with purified water;
[0017] More preferably, the treatment includes: washing with sodium chloride solution, treating with hydrochloric acid, washing with purified water, treating with SDS solution, and washing with purified water again.
[0018] Preferably, the animal tissue includes pig skin and / or the mucosal layer of pig small intestine; more preferably, the animal tissue is either pig skin or the mucosal layer of pig small intestine.
[0019] Preferably, the sodium chloride solution is a 0.1%-3% sodium chloride solution; more preferably, the sodium chloride solution is a 0.9% sodium chloride solution.
[0020] Preferably, the hydrochloric acid is 1%-5% hydrochloric acid; more preferably, the hydrochloric acid is 2% hydrochloric acid.
[0021] Preferably, the SDS solution is a 1%-5% SDS solution; more preferably, the SDS solution is a 1% SDS solution.
[0022] Preferably, the at least three transparency operations include five transparency operations;
[0023] More preferably, the at least three transparency operations include four transparency operations;
[0024] More preferably, the at least three transparency processes include three transparency processes;
[0025] Most preferably, the at least three transparency processes include a first transparency process, a second transparency process, and a third transparency process.
[0026] Preferably, the first transparentization includes: hydrating the dry decellularized collagen membrane obtained in step (a) to make it transparent, and treating it with sodium hydroxide solution.
[0027] Preferably, the second transparency process includes: rinsing with purified water followed by soaking in an acetic acid solution.
[0028] Preferably, the third transparency process includes: after rinsing with purified water, heating the collagen membrane in a water bath, and then rapidly cooling the collagen membrane in a sodium hydroxide solution.
[0029] Preferably, step (b) further includes: washing the cooled collagen membrane with purified water, adding purified water until it covers the collagen membrane, adjusting the pH value with hydrochloric acid, and washing it again with purified water to obtain a transparent collagen membrane.
[0030] Preferably, the sodium hydroxide solution is a 0.3%-10% sodium hydroxide solution; more preferably, the sodium hydroxide solution is a 1% sodium hydroxide solution.
[0031] Preferably, the acetic acid solution is a 1%-5% acetic acid solution; more preferably, the acetic acid solution is a 1% acetic acid solution.
[0032] Preferably, the water bath heating is performed by heating in water at 30℃-70℃ for 5-15 minutes; more preferably, the water bath heating is performed by heating in water at 50℃ for 10 minutes.
[0033] Preferably, the sodium hydroxide solution is a sodium hydroxide solution at 0℃-20℃; more preferably, the sodium hydroxide solution is a sodium hydroxide solution at 0℃.
[0034] Preferably, the hydrochloric acid is 0.1%-10% hydrochloric acid; more preferably, the hydrochloric acid is 1% hydrochloric acid.
[0035] Preferably, the pH adjustment is to adjust the pH value to 6-7; more preferably, the pH adjustment is to adjust the pH value to 7.
[0036] The present invention also provides a collagen membrane prepared according to the method, wherein the collagen membrane is a transparent collagen membrane.
[0037] The present invention also provides the application of the collagen membrane prepared according to the method in the preparation of tissue and organ repair products for humans or animals.
[0038] Preferably, the organ is the cornea.
[0039] The effects of the invention
[0040] This invention provides a method for preparing a transparent collagen membrane. The method uses animal-derived materials and obtains the collagen membrane through decellularization, making its triple helix structure more closely resemble the tissue itself, resulting in good mechanical and degradation properties. The collagen membrane prepared according to this invention, after three transparentization treatments, exhibits a uniform collagen network structure and a stable, uniformly transparent color. Its translucency is not affected by pH changes, enabling it to accelerate wound healing while maintaining the aesthetic appearance of the healed tissue. Attached Figure Description
[0041] Figure 1 This data represents the change in product transmittance with each processing step.
[0042] Figure 2 This data shows the change in the transmittance of the treated product as a function of pH value when the product is immersed in solutions of different pH values.
[0043] Figure 3 It is a decellularized collagen membrane of hydrated porcine small intestinal mucosa.
[0044] Figure 4 The image shows a transparent porcine small intestinal mucosa decellularized collagen membrane. (A) The transparent state of the transparent porcine small intestinal mucosa decellularized collagen membrane placed on a culture dish; (B) The transparent state of the transparent porcine small intestinal mucosa decellularized collagen membrane placed on a glove.
[0045] Figure 5 It is a decellularized collagen membrane of hydrated porcine dermal matrix.
[0046] Figure 6 The transparent porcine dermal matrix decellularized collagen membrane. (A) The transparent state of the transparent porcine dermal matrix decellularized collagen membrane placed on a culture dish; (B) The transparent state of the transparent porcine dermal matrix decellularized collagen membrane placed on a glove. Detailed Implementation
[0047] To make the technical solution and beneficial effects of the present invention more apparent and understandable, a detailed description is provided below by listing specific embodiments. The accompanying drawings are not necessarily drawn to scale, and local features may be enlarged or reduced to more clearly show the details of the local features; unless otherwise defined, the technical and scientific terms used herein have the same meanings as those in the technical field to which this application pertains.
[0048] Unless otherwise specified, the experimental methods used in this invention are conventional methods. Unless otherwise specified, the materials, reagents, and standards used in the following examples are commercially available.
[0049] As used in this article, the term "decellularization" refers to the process of decellularizing tissues / organs using appropriate methods to remove cells and other antigen molecules that may cause rejection reactions, while retaining the 3D structure (collagen and elastin) and functional proteins (such as glucan, proteoglycans, and growth factors). This process results in a novel biomaterial with bio-inducible functions that can form specific functionalized tissues in vivo / in vitro and can be used for tissue damage repair / reconstruction.
[0050] As used in this article, the term "collagen membrane" refers to a membrane or membrane-like product made primarily of collagen through decellularization or other methods.
[0051] As used in this article, the term "animal-derived" refers to something obtained from an animal, such as a pig, a cow, or a sheep.
[0052] As used in this article, the term "purified water" refers to deionized water or deeply desalinated water.
[0053] As used in this article, the term "SDS" refers to sodium dodecyl sulfate.
[0054] As used herein, the terms “comprising” or “including” mean that the method, structure, or composition includes any example of a step / operation, component, or part, but does not exclude any other step / operation, component, or part.
[0055] In the context of this invention, when referring to numerical ranges, it should be understood that the upper and lower limits of the range, each intermediate value in between, and any other specified value or intermediate value within the range are included within this invention. The upper and lower limits of these smaller ranges that may be independently included within the range are also included within this invention, subject to any explicit exclusions from the specified range. When a specified range includes one or both of the limit values, the range excluding either of the limit values is also within this invention.
[0056] This invention provides a method for preparing a transparent collagen membrane, wherein the method includes the following steps:
[0057] (a) Obtaining decellularized collagen membranes of animal origin;
[0058] (b) Transparent the collagen membrane at least three times.
[0059] In some implementations, step (a) includes:
[0060] (a1) Take animal tissue and remove the surface fat and impurities;
[0061] (a2) Processing animal tissues after removing surface fat and impurities;
[0062] (a3) The animal tissue treated in step (a2) was freeze-dried using a freeze dryer to obtain a dry decellularized collagen membrane.
[0063] In some embodiments, the treatment includes washing with a sodium chloride solution.
[0064] In some embodiments, the treatment includes: washing with a sodium chloride solution, treating with hydrochloric acid, and then washing with purified water.
[0065] In some embodiments, the treatment includes: washing with sodium chloride solution, treating with hydrochloric acid, washing with purified water, treating with SDS solution, and washing with purified water again.
[0066] In some embodiments, the animal tissue includes porcine skin and / or porcine small intestinal mucosa; more preferably, the animal tissue is either porcine skin or porcine small intestinal mucosa.
[0067] In some implementations, step (a1) includes taking pig skin, removing the fat layer and the epidermis to obtain the dermis.
[0068] In some implementations, step (a1) includes taking a mucosal layer of the pig small intestine and removing surface fat and impurities.
[0069] In some embodiments, the sodium chloride solution is a 0.1%-3% sodium chloride solution.
[0070] In some embodiments, the sodium chloride solution is a 0.9% sodium chloride solution.
[0071] In some embodiments, the cleaning with sodium chloride solution involves cleaning four times with a 0.1%-3% sodium chloride solution.
[0072] In some embodiments, the cleaning with sodium chloride solution involves cleaning four times with a 0.9% sodium chloride solution.
[0073] In some embodiments, the cleaning with sodium chloride solution is performed using an electric stirrer with a rotation speed of 100 rpm.
[0074] In some implementations, the cleaning with sodium chloride solution is performed for 30 minutes each time.
[0075] In some embodiments, the hydrochloric acid is 1%-5% hydrochloric acid.
[0076] In some embodiments, the hydrochloric acid is 2% hydrochloric acid.
[0077] In some embodiments, the hydrochloric acid treatment is a treatment with 1%-5% hydrochloric acid for 2 hours.
[0078] In some embodiments, the hydrochloric acid treatment is a treatment with 2% hydrochloric acid for 2 hours.
[0079] In some embodiments, the hydrochloric acid treatment is performed using an electric stirrer with a rotation speed of 100 rpm.
[0080] In some embodiments, the SDS solution is a 1%-5% SDS solution.
[0081] In some embodiments, the SDS solution is a 1% SDS solution.
[0082] In some embodiments, the treatment with SDS solution is to treat with 1%-5% SDS solution for 24 hours.
[0083] In some embodiments, the treatment with SDS solution is to treat with 1% SDS solution for 24 hours.
[0084] In some embodiments, the SDS solution treatment is performed using an electric stirrer with a rotation speed of 100 rpm.
[0085] In some implementations, the washing with purified water involves washing three times with purified water.
[0086] In some embodiments, the washing with purified water is performed using an electric stirrer with a rotation speed of 100 rpm.
[0087] In some implementations, the rinsing with purified water is performed for 30 minutes each time.
[0088] In some implementations, the freeze-drying is performed using a freeze dryer for 16 hours.
[0089] In some implementations, the at least three transparencys include five transparencys.
[0090] In some implementations, the at least three transparencys include four transparencys.
[0091] In some implementations, the at least three transparencys include three transparencys.
[0092] In some implementations, the at least three transparency processes include a first transparency process, a second transparency process, and a third transparency process.
[0093] In some embodiments, the first transparency process includes: hydrating the dry, decellularized collagen membrane prepared in step (a) to make it transparent, and treating it with a sodium hydroxide solution.
[0094] In some embodiments, the second transparency process includes rinsing with purified water followed by immersion in an acetic acid solution.
[0095] In some embodiments, the third transparency process includes: after rinsing with purified water, heating the collagen membrane in a water bath, and then rapidly cooling the collagen membrane in a sodium hydroxide solution.
[0096] In some embodiments, step (b) further includes: washing the cooled collagen membrane with purified water, adding purified water until it covers the collagen membrane, adjusting the pH value with hydrochloric acid, and washing it again with purified water to obtain a transparent collagen membrane.
[0097] In some embodiments, the sodium hydroxide solution is a 0.3%-10% sodium hydroxide solution.
[0098] In some embodiments, the sodium hydroxide solution is a 1% sodium hydroxide solution.
[0099] In some embodiments, the treatment with sodium hydroxide solution is to treat with a 0.3%-10% sodium hydroxide solution for 1 hour.
[0100] In some embodiments, the treatment with sodium hydroxide solution is to treat with a 1% sodium hydroxide solution for 1 hour.
[0101] In some embodiments, the treatment with sodium hydroxide solution is performed using an electric stirrer with a rotation speed of 60 rpm.
[0102] In some embodiments, the acetic acid solution is a 1%-5% acetic acid solution.
[0103] In some embodiments, the acetic acid solution is a 1% acetic acid solution.
[0104] In some embodiments, the immersion treatment with acetic acid solution is a immersion treatment with 1%-5% acetic acid solution for 2 hours.
[0105] In some embodiments, the immersion treatment with acetic acid solution is a immersion treatment with 1% acetic acid solution for 2 hours.
[0106] In some embodiments, the water bath heating is performed by heating in water at 30°C-70°C for 5-15 minutes.
[0107] In some implementations, the water bath heating is performed by heating in water at 50°C for 10 minutes.
[0108] In some embodiments, the sodium hydroxide solution is a sodium hydroxide solution at 0°C-20°C.
[0109] In some embodiments, the sodium hydroxide solution is a sodium hydroxide solution at 0°C.
[0110] In some embodiments, the hydrochloric acid is 0.1%-10% hydrochloric acid.
[0111] In some embodiments, the hydrochloric acid is 1% hydrochloric acid.
[0112] In some implementations, the pH adjustment is to adjust the pH to 6-7.
[0113] In some implementations, the pH adjustment is to adjust the pH to 7.
[0114] In some implementations, the washing with purified water involves washing three times with purified water.
[0115] In some embodiments, the washing with purified water is performed using an electric stirrer with a rotation speed of 100 rpm.
[0116] In some implementations, the rinsing with purified water is performed for 30 minutes each time.
[0117] The present invention also provides a collagen membrane prepared according to the method, wherein the collagen membrane is a transparent collagen membrane.
[0118] In some embodiments, the collagen membrane has a light transmittance of 77%-87%.
[0119] In some embodiments, the collagen membrane has a light transmittance of 87%.
[0120] The present invention also provides the application of the collagen membrane prepared according to the method in the preparation of tissue and organ repair products for humans or animals.
[0121] In some implementations, the organ is the cornea.
[0122] In some implementations, the tissue / organ repair product is a product for repairing wound areas.
[0123] In some implementations, the tissue / organ repair product is a patch-like material.
[0124] In some embodiments, the tissue / organ repair product is a patch-like material used for repairing wound areas after corneal dermoid tumor resection.
[0125] Example 1: Clearing treatment of decellularized porcine dermal matrix
[0126] 1. Obtaining animal-derived decellularized collagen membranes
[0127] (1) Take pig skin raw material, remove the fat layer and epidermis to obtain the dermis, and use a light transmittance tester to test the light transmittance of the dermis.
[0128] (2) Wash with 0.9% sodium chloride solution 4 times, with the electric stirrer speed at 150 rpm, each wash for 30 minutes;
[0129] (3) Treat the leather in a beaker with 2% hydrochloric acid for 2 hours, with the electric stirrer speed at 150 rpm;
[0130] (4) Rinse with purified water 3 times, with the electric stirrer speed at 150 rpm, each rinse lasting 30 minutes;
[0131] (5) Treat with 1% SDS solution for 24 hours, with electric stirrer speed of 150 rpm;
[0132] (6) Rinse with purified water 3 times, with the electric stirrer speed at 150 rpm, each rinse lasting 30 minutes;
[0133] (7) Use a freeze dryer to freeze dry for 24 hours to obtain a dry collagen membrane, which is white at this time.
[0134] 2. Collagen membrane becomes transparent
[0135] (1) Hydrate the dried collagen membrane until it becomes transparent (e.g.) Figure 5 As shown), it was treated with 2% sodium hydroxide solution for 2 hours, with the electric stirrer speed at 100 rpm (first transparentization);
[0136] (2) Wash with purified water 3 times, with an electric stirrer speed of 150 rpm, each wash for 30 minutes, and use a light transmittance tester to test the light transmittance of the dermis.
[0137] (3) Soak in 1% acetic acid solution for 3 hours (second transparentization);
[0138] (4) Wash with purified water 3 times, with an electric stirrer speed of 150 rpm, each wash lasting 30 minutes. Use a light transmittance tester to test the light transmittance of the dermis.
[0139] (5) Heat the collagen membrane in a 50°C water bath for 10 minutes;
[0140] (6) Place the collagen membrane in a 0℃ sodium hydroxide solution and cool it rapidly for 5 minutes (third transparentization);
[0141] (7) Rinse with purified water 3 times, with the electric stirrer speed at 150 rpm, each rinse lasting 30 minutes;
[0142] (8) Add purified water until it covers the collagen membrane, adjust the pH to 6.7 with hydrochloric acid, and soak for 1 hour;
[0143] (9) Wash three times with purified water, using an electric stirrer at 150 rpm for 30 minutes each time. After washing, a transparent collagen membrane is obtained (e.g., ...). Figure 6 (As shown).
[0144] The change in product light transmittance with processing steps is as follows: Figure 1 As shown, the light transmittance of the dermal matrix reached 56% after the first transparency treatment, 61% after the second transparency treatment, and 77% after the third transparency treatment.
[0145] 3. Transmittance testing of products at different pH values
[0146] The transparent collagen membrane was immersed in solutions with pH values of 1, 2, 3, 4, 5, 6, 7, and 8, respectively, for 5 minutes. Subsequently, the transmittance of the dermis was tested using a transmittance meter. The change in transmittance of the product with pH is shown below. Figure 2 As shown, the transmittance is lower when the pH value is lower and higher when the pH value is closer to neutral, with a relatively small overall variation.
[0147] Example 2: Clearing treatment of decellularized matrix in porcine small intestinal mucosa
[0148] 1. Obtaining animal-derived decellularized collagen membranes
[0149] (1) Take the mucosal layer of pig small intestine, remove the surface fat and impurities, and use a light transmittance tester to test the light transmittance of the dermis layer;
[0150] (2) Wash with 0.9% sodium chloride solution 4 times, with the electric stirrer speed at 100 rpm, each wash for 30 minutes;
[0151] (3) Treat the small intestinal mucosa in a beaker with 1% hydrochloric acid for 2 hours, with the electric stirrer speed at 100 rpm;
[0152] (4) Rinse with purified water 3 times, with the electric stirrer speed at 100 rpm, and each rinse for 30 minutes;
[0153] (5) Treat with 1% SDS solution for 24 hours, with electric stirrer speed of 100 rpm;
[0154] (6) Rinse with purified water 3 times, with the electric stirrer speed at 100 rpm, each rinse lasting 30 minutes;
[0155] (7) Use a freeze dryer to freeze dry for 16 hours to obtain a dry collagen membrane, which is white at this time.
[0156] 2. Collagen membrane becomes transparent
[0157] (1) Hydrate the dried collagen membrane until it becomes transparent (e.g.) Figure 3 As shown), it was treated with 1% sodium hydroxide solution for 1 hour, with the electric stirrer speed at 60 rpm (first transparentization);
[0158] (2) Wash with purified water 3 times, with an electric stirrer speed of 100 rpm, each wash for 30 minutes, and use a light transmittance tester to test the light transmittance of the dermis.
[0159] (3) Soak in 1% acetic acid solution for 2 hours (second transparentization);
[0160] (4) Wash with purified water 3 times, with the electric stirrer speed at 100 rpm, each wash for 30 minutes, and use a light transmittance tester to test the light transmittance of the dermis.
[0161] (5) Heat the collagen membrane in a 50°C water bath for 10 minutes;
[0162] (6) Place the collagen membrane in a 0℃ sodium hydroxide solution and cool it rapidly for 5 minutes (third transparentization);
[0163] (7) Rinse with purified water 3 times, with the electric stirrer speed at 100 rpm, each rinse lasting 30 minutes;
[0164] (8) Add purified water until it covers the collagen membrane, adjust the pH to 6.7 with hydrochloric acid, and soak for 1 hour;
[0165] (9) Wash three times with purified water, using an electric stirrer at 100 rpm for 30 minutes each time. A transparent collagen membrane (e.g.) will be obtained after washing. Figure 4 As shown, a transmittance tester was used to test the transmittance of the dermis.
[0166] The change in product light transmittance with processing steps is as follows: Figure 1 As shown, the light transmittance of the dermal matrix reached 68% after the first transparency treatment, 71% after the second transparency treatment, and 87% after the third transparency treatment.
[0167] 3. Transmittance testing of products at different pH values
[0168] The transparent collagen membrane was immersed in solutions with pH values of 1, 2, 3, 4, 5, 6, 7, and 8, respectively, for 5 minutes. Subsequently, the transmittance of the dermis was tested using a transmittance meter. The change in transmittance of the product with pH is shown below. Figure 2 As shown, the transmittance is lower when the pH value is lower and higher when the pH value is closer to neutral, with a relatively small overall variation.
[0169] It should be understood that the above embodiments are exemplary and are not intended to encompass all possible implementations included in the claims. Various modifications and changes can be made to the above embodiments without departing from the scope of this disclosure. Similarly, the various technical features of the above embodiments can be arbitrarily combined to form other embodiments of the present invention that may not be explicitly described. Therefore, the above embodiments only illustrate several implementations of the present invention and do not limit the scope of protection of this patent.
Claims
1. A method for preparing a transparent collagen membrane, characterized in that, The method includes the following steps: (a) Obtaining animal-derived decellularized collagen membranes; (b) Transparentify the collagen membrane at least three times; The at least three transparency processes include a first transparency, a second transparency, and a third transparency. The first transparentization includes: hydrating the dry decellularized collagen membrane obtained in step (a) to make it transparent, and treating it with sodium hydroxide solution; The second transparency process includes: rinsing with purified water followed by soaking in an acetic acid solution; The third transparency process includes: after rinsing with purified water, heating the collagen membrane in a water bath, and then rapidly cooling the collagen membrane in a sodium hydroxide solution. The sodium hydroxide solution is a 0.3%-10% sodium hydroxide solution; The acetic acid solution is a 1%-5% acetic acid solution; The water bath heating is performed by heating in water at 30℃-70℃ for 5-15 minutes; The sodium hydroxide solution is a sodium hydroxide solution with a temperature of 0℃-20℃.
2. The method according to claim 1, characterized in that, Step (a) includes: (a1) Take animal tissue and remove surface fat and impurities; (a2) Processing animal tissues after removing surface fat and impurities; (a3) The animal tissue treated in step (a2) was freeze-dried using a freeze dryer to obtain a decellularized collagen membrane in a dry state.
3. The method according to claim 2, characterized in that, The treatment described in step (a2) includes: washing with a sodium chloride solution.
4. The method according to claim 3, characterized in that, The treatment described in step (a2) includes: washing with sodium chloride solution, treating with hydrochloric acid, and then washing with purified water.
5. The method according to claim 4, characterized in that, The treatment described in step (a2) includes: washing with sodium chloride solution, treating with hydrochloric acid, washing with purified water, treating with SDS solution, and washing with purified water.
6. The method according to claim 2, characterized in that: The animal tissues include pig skin and / or the mucosal layer of pig small intestine.
7. The method according to claim 6, characterized in that: The animal tissue is either pig skin or the mucosal layer of pig small intestine.
8. The method according to any one of claims 3-5, characterized in that: The sodium chloride solution is a 0.1%-3% sodium chloride solution.
9. The method according to claim 8, characterized in that: The sodium chloride solution is a 0.9% sodium chloride solution.
10. The method according to any one of claims 4-5, characterized in that: The hydrochloric acid is 1%-5% hydrochloric acid.
11. The method according to claim 10, characterized in that: The hydrochloric acid is 2% hydrochloric acid.
12. The method according to claim 5, characterized in that: The SDS solution is a 1%-5% SDS solution.
13. The method according to claim 12, characterized in that: The SDS solution is a 1% SDS solution.
14. The method according to claim 1, characterized in that, Step (b) further includes: washing the cooled collagen membrane with purified water, adding purified water until it covers the collagen membrane, adjusting the pH value with hydrochloric acid, and washing it again with purified water to obtain a transparent collagen membrane.
15. The method according to claim 1, characterized in that: The sodium hydroxide solution is a 1% sodium hydroxide solution.
16. The method according to claim 1, characterized in that: The acetic acid solution is a 1% acetic acid solution.
17. The method according to claim 1, characterized in that: The water bath heating involves heating in water at 50°C for 10 minutes.
18. The method according to claim 1, characterized in that: The sodium hydroxide solution is a sodium hydroxide solution at 0°C.
19. The method according to claim 14, characterized in that: The hydrochloric acid is 0.1%-10% hydrochloric acid.
20. The method according to claim 19, characterized in that: The hydrochloric acid is 1% hydrochloric acid.
21. The method according to claim 14, characterized in that: The pH adjustment is to adjust the pH value to 6-7.
22. The method according to claim 21, characterized in that: The pH adjustment is to adjust the pH value to 7.
23. A collagen membrane prepared according to the method of claim 1, characterized in that, The collagen membrane is a transparent collagen membrane.
24. The application of a collagen membrane prepared according to claim 1 in the preparation of tissue and organ repair products for humans or animals, characterized in that, The organ in question is the cornea.