Porcine-derived extracellular matrix material and methods and uses for removing immunogenicity thereof

Abstract

The application discloses a porcine-derived extracellular matrix material and a method and application for removing immunogenicity of the porcine-derived extracellular matrix. The method comprises the following steps: ultrasonic treatment of porcine-derived fascial tissue in a first to sixth mixed solution, a first ethanol solution and pure water in sequence, and then biological enzyme hydrolysis; the first mixed solution comprises 0.8-1.2% triton X-100, 5-10% acetone and 15-25% anhydrous ethanol; the second mixed solution comprises 1.5-2.0% triton X-100, 2.0-3.0% sodium carbonate and 70-76.5% anhydrous ethanol; the third mixed solution comprises 2.0-3.0% SDS, 2.0-3.0% citric acid and 20-30% anhydrous ethanol; the fourth mixed solution comprises 1.0-2.0% SDS, 2.0-3.0% acetic acid and 20-30% anhydrous ethanol; the fifth mixed solution is a PBS solution; and the sixth mixed solution is a sodium bicarbonate aqueous solution.

Description

Technical Field

[0001] This application relates to the field of biomaterials technology, specifically to a porcine extracellular matrix material and a method and application for removing the immunogenicity of porcine extracellular matrix. Background Technology

[0002] Biomaterials composed of extracellular matrix (ECM) are typically derived from decellularized tissues or organs. Tissues are composed of cells and extracellular matrix (ECM), and their cellular components, debris, DNA, and other elements are highly immunogenic. Direct implantation into the human body can trigger inflammation or immune rejection, leading to transplant failure. In contrast, ECM is a complex complex of structural and functional proteins, and its components are generally protected and well-tolerated across different species. Decellularized tissues and organs are widely used in tissue engineering and regenerative medicine.

[0003] Porcine endocrine disruptor membranes (ECMs) are considered ideal biomaterials due to their wide availability and structural similarity to human tissues. Thorough removal of these immunogenic components is a prerequisite for the clinical application of porcine ECMs. The ideal decellularization technique preserves the complete components and three-dimensional ultrastructure of the ECM.

[0004] Currently, the mainstream method for removing immunogenicity from ECM is a combination of physical and chemical methods for decellularization. However, existing technologies have significant shortcomings: First, the decellularization process is often incomplete, resulting in high levels of residual cells and DNA, which still poses a risk of triggering an immune response. Second, the large quantities or repeated use of chemical reagents to ensure the decellularization effect can easily leave residues inside the ECM scaffold. These residual chemical reagents may be toxic or irritating to the recipient tissue, and some sensitive individuals may experience adverse reactions. Finally, many methods are complex and require stringent conditions, making it difficult to scale up to industrial production. Summary of the Invention

[0005] To address the aforementioned deficiencies in this field, this application aims to provide a porcine extracellular matrix material and a method and application for removing the immunogenicity of porcine extracellular matrix.

[0006] According to one aspect of this application, a method for removing porcine extracellular matrix immunogenicity is provided, comprising:

[0007] Porcine fascia tissue was pretreated.

[0008] The pretreated porcine fascia tissue was subjected to a first ultrasonic treatment in a first mixed solution to obtain a first intermediate product.

[0009] The first intermediate product was subjected to a second ultrasonic treatment in the second mixed solution to obtain the second intermediate product.

[0010] The second intermediate product was subjected to a third ultrasonic treatment in the third mixed solution to obtain the third intermediate product.

[0011] The third intermediate product was subjected to a fourth ultrasonic treatment in the fourth mixed solution to obtain the fourth intermediate product.

[0012] The fourth intermediate product was subjected to a fifth ultrasonic treatment in the fifth mixed solution to obtain the fifth intermediate product.

[0013] The fifth intermediate product was subjected to a sixth ultrasonic treatment in the sixth mixed solution to obtain the sixth intermediate product.

[0014] The sixth intermediate product was sequentially immersed in a first ethanol solution and pure water for a seventh and eighth ultrasonic treatment to obtain the seventh intermediate product.

[0015] The seventh intermediate was subjected to enzymatic hydrolysis to obtain a porcine extracellular matrix that had been de-immunogenic.

[0016] The first mixed solution is prepared by mixing the following raw materials in parts by weight: Triton X-100: 0.8~1.2%; acetone: 5~10%; anhydrous ethanol: 15~25%; water balance;

[0017] The second mixed solution is prepared by mixing the following raw materials in parts by weight: Triton X-100: 1.5~2.0%; sodium carbonate: 2.0~3.0%; anhydrous ethanol: 70~76.5%; water balance;

[0018] The third mixed solution is prepared by mixing the following raw materials in parts by weight: sodium dodecyl sulfate: 2.0~3.0%; citric acid: 2.0~3.0%; anhydrous ethanol: 20~30%; water balance;

[0019] The fourth mixed solution is prepared by mixing the following raw materials in parts by weight: sodium dodecyl sulfate: 1.0~2.0%; acetic acid: 2.0~3.0%; anhydrous ethanol: 20~30%; water balance;

[0020] The fifth mixed solution is a PBS solution with a pH of 7.0 to 7.5; the sixth mixed solution is an aqueous solution of sodium bicarbonate.

[0021] According to some embodiments of this application, the pretreatment includes: immersing the porcine fascia tissue in a fat-soluble solvent, subjecting it to ultrasonic treatment for 1-2 hours, draining the solvent, and rinsing off any remaining solvent with purified water.

[0022] According to some embodiments of this application, the first ethanol solution is an ethanol solution with a mass fraction of 5-15%.

[0023] According to some embodiments of this application, the duration of the first ultrasonic treatment is 1 to 3 hours;

[0024] The second ultrasound treatment takes 1 to 3 hours;

[0025] The third ultrasound treatment takes 1 to 3 hours;

[0026] The fourth ultrasound treatment takes 1 to 3 hours;

[0027] The fifth ultrasound treatment takes 1-2 hours;

[0028] The sixth ultrasound treatment takes 1-2 hours;

[0029] The seventh ultrasound treatment takes 1 to 2 hours.

[0030] According to some embodiments of this application, the amounts of the first mixed solution, the second mixed solution, the third mixed solution, and the fourth mixed solution are not less than 7 times the mass of the pretreated porcine fascia tissue;

[0031] The amount of the fifth mixed solution shall not be less than five times the mass of the first intermediate product;

[0032] The amount of the first ethanol solution used shall not be less than 5 times the mass of the second intermediate product;

[0033] The amount of pure water used shall not be less than five times the mass of the second intermediate product.

[0034] According to some embodiments of this application, the mass concentration of the sodium bicarbonate aqueous solution is 2% to 4%.

[0035] According to some embodiments of this application, the fat-soluble solvent is one or more of acetone, n-hexane, and ethyl acetate.

[0036] According to some embodiments of this application, the biological enzyme includes one or more of trypsin, pepsin, and collagenase.

[0037] According to another aspect of this application, a porcine extracellular matrix material prepared by the above method is also provided, wherein the porcine extracellular matrix material contains ≤2 residual cells / field of view, ≤5 μg / g residual DNA, ≤10 μg / g residual Triton X-100, and ≤3 mg / g residual sodium dodecyl sulfate.

[0038] According to one aspect of this application, the porcine extracellular matrix material prepared by the above method is used in tissue engineering scaffolds, wound dressings, or biological patches.

[0039] Compared with the prior art, this application has at least the following beneficial effects:

[0040] This application provides a method for removing porcine extracellular matrix immunogenicity, which can efficiently and thoroughly remove immunogenicity while ensuring extremely low chemical reagent residues, and the process is suitable for industrial production. Detailed Implementation

[0041] The technical solution of this application will be clearly and completely described below with reference to the embodiments of this application. Obviously, the described embodiments are only some embodiments of this application, not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0042] It should be particularly noted that similar substitutions and modifications made to this application are obvious to those skilled in the art, and they are all considered to be included in this application. Those skilled in the art can obviously make modifications or appropriate alterations and combinations to the methods and applications described herein without departing from the content, spirit, and scope of this application to implement and apply the technology of this application. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments.

[0043] Unless otherwise specified, this application is conducted under standard conditions or conditions recommended by the manufacturer. The raw materials or excipients used, as well as the reagents or instruments used, whose manufacturers are not specified, are all conventional products that can be obtained commercially.

[0044] The following is a detailed description of this application.

[0045] To address the significant shortcomings of existing technologies in removing ECM immunogenicity, this application provides a porcine extracellular matrix material, a method for removing porcine extracellular matrix immunogenicity, and its application.

[0046] In some examples, the method for removing porcine extracellular matrix immunogenicity according to this application includes:

[0047] Porcine fascia tissue was immersed in a lipid-soluble solvent and sonicated for 1-2 hours. The solvent was drained, and residual solvent was rinsed with purified water. The pretreated porcine fascia tissue was then immersed in a first mixed solution for a first sonication for 1-3 hours to obtain a first intermediate product. The first intermediate product was then immersed in a second mixed solution for a second sonication for 1-3 hours to obtain a second intermediate product. The second intermediate product was then sonicated in a third mixed solution for a third sonication to obtain a third intermediate product. The third intermediate product was then sonicated in a fourth mixed solution for a fourth sonication to obtain a fourth intermediate product. The fourth intermediate product was then sonicated in a fifth mixed solution for a fifth sonication to obtain a fifth intermediate product. The fifth intermediate product was then sonicated in a sixth mixed solution for a sixth sonication to obtain a sixth intermediate product. The sixth intermediate product was then immersed in a first ethanol solution and purified water for a seventh and eighth sonication treatment to obtain a seventh intermediate product. The seventh intermediate product was then enzymatically hydrolyzed, centrifuged, and freeze-dried to obtain ECM material with low immunogenicity.

[0048] The first mixed solution is prepared by mixing the following raw materials in parts by weight:

[0049] Triton X-100: 0.8~1.2%;

[0050] Acetone: 5-10%;

[0051] Anhydrous ethanol: 15~25%;

[0052] Water balance;

[0053] The second mixed solution is prepared by mixing the following raw materials in parts by weight:

[0054] Triton X-100: 1.5~2.0%;

[0055] Sodium carbonate: 2.0~3.0%;

[0056] Anhydrous ethanol: 70~76.5%;

[0057] Water balance;

[0058] The third mixed solution is prepared by mixing the following raw materials in parts by weight:

[0059] Sodium dodecyl sulfate: 2.0~3.0%;

[0060] Citric acid: 2.0~3.0%;

[0061] Anhydrous ethanol: 20~30%;

[0062] Water balance;

[0063] The fourth mixed solution is prepared by mixing the following raw materials in parts by weight:

[0064] Sodium dodecyl sulfate: 1.0~2.0%;

[0065] Acetic acid: 2.0~3.0%;

[0066] Anhydrous ethanol: 20~30%;

[0067] Water balance;

[0068] The fifth mixed solution is a PBS solution with a pH of 7.0–7.5;

[0069] The sixth mixed solution is an aqueous solution of sodium bicarbonate, optionally with a concentration of 2% to 4%, optionally 3%.

[0070] In some examples, the duration of the first ultrasound treatment was 1–3 hours; the duration of the second ultrasound treatment was 1–3 hours; the duration of the third ultrasound treatment was 1–3 hours; the duration of the fourth ultrasound treatment was 1–3 hours; the duration of the fifth ultrasound treatment was 1–2 hours; the duration of the third ultrasound treatment was 1–2 hours; and the duration of the fourth ultrasound treatment was 1–2 hours.

[0071] In some examples, the amount of lipid-soluble solution used is not less than 5 times the tissue mass; the amount of first ethanol used is not less than 5 times the tissue mass; and the amount of pure water used is not less than 5 times the tissue mass. The amount of first mixed solution used is not less than 7 times the mass of pretreated porcine fascia tissue; the amount of second mixed solution used is not less than 7 times the mass of first intermediate product; the amount of third mixed solution used is not less than 7 times the mass of first intermediate product; and the amount of fourth mixed solution used is not less than 7 times the mass of first intermediate product.

[0072] In some examples, the lipid-soluble solvent is one or more of acetone, n-hexane, and ethyl acetate; the biological enzyme is one or more of trypsin, pepsin, and collagenase.

[0073] In some examples, the porcine extracellular matrix material of this application has the following specifications: residual cells ≤2 per field of view (slide staining method), residual DNA ≤5μg / g, residual Triton X-100 ≤10μg / g, and residual sodium dodecyl sulfate ≤3mg / g.

[0074] The technical solution of this application will be further described below with reference to specific embodiments.

[0075] Example 1

[0076] Take 100g of abdominal wall fascia tissue from a healthy pig, cut it into small pieces, soak it in 500mL of acetone, sonicate it (300W power, 40kHz frequency) for 1.5 hours, drain it, and rinse it three times with 1000mL of purified water.

[0077] The tissue was immersed in 700 mL of the first mixed solution (Triton X-100: acetone: anhydrous ethanol: water = 1:7:20:72 (mass ratio)), sonicated for 2 hours, and then drained.

[0078] The tissue was immersed in 700 mL of a second mixed solution (Triton X-100: sodium carbonate: anhydrous ethanol: water = 1.8: 2.5: 72.7: 23 (mass ratio)), sonicated for 2 hours, and then drained.

[0079] The tissue was immersed in 700 mL of a third mixed solution (sodium dodecyl sulfate: citric acid: anhydrous ethanol: water = 2.5: 2.5: 25: 70 (mass ratio)), sonicated for 2 hours, and then drained.

[0080] The tissue was immersed in 700 mL of the fourth mixed solution (sodium dodecyl sulfate: acetic acid: anhydrous ethanol: water = 1.5: 2.5: 25: 71 (mass ratio)), sonicated for 2 hours, and then drained.

[0081] Immerse the tissue in 500ml of purified water, add PBS solution, adjust the pH to 7.0-7.5, sonicate for 1.5h, and drain.

[0082] The tissue was immersed in a 3% sodium bicarbonate aqueous solution, sonicated for 1.5 hours, and then drained.

[0083] The tissue was immersed in 500 ml of 10% ethanol, sonicated for 1.5 h, and then drained.

[0084] The tissue was immersed in 500ml of purified water, sonicated for 1.5h, and drained; the tissue was then freeze-dried to obtain the dried product.

[0085] Take 5g of dried product and enzymatically hydrolyze it with 500ml of pepsin solution (0.1%, pH2) at room temperature for 12 hours. Centrifuge and discard the supernatant. Freeze-dry the precipitate to obtain white flocculent ECM material.

[0086] Example 2

[0087] Take 100g of abdominal wall fascia tissue from a healthy pig, cut it into small pieces, soak it in 500mL of ethyl acetate, sonicate it (300W power, 40kHz frequency) for 2 hours, drain it, and rinse it 3 times with 1000mL of purified water.

[0088] The tissue was immersed in 700 mL of the first mixed solution (Triton X-100: acetone: anhydrous ethanol: water = 0.8:5:15:79.2 (mass ratio)), sonicated for 3 hours, and then drained.

[0089] The tissue was immersed in 700 mL of a second mixed solution (Triton X-100: sodium carbonate: anhydrous ethanol: water = 1.5: 2.0: 76.5: 20 (mass ratio)), sonicated for 3 hours, and then drained.

[0090] The tissue was immersed in 700 mL of a third mixed solution (sodium dodecyl sulfate: citric acid: anhydrous ethanol: water = 2:2:20:76 (mass ratio)), sonicated for 1 hour, and then drained.

[0091] The tissue was immersed in 700 mL of the fourth mixed solution (sodium dodecyl sulfate: acetic acid: anhydrous ethanol: water = 1:2:20:77 (mass ratio)), sonicated for 1.5 hours, and then drained.

[0092] Immerse the tissue in 500ml of purified water, add PBS solution, adjust the pH to 7.0-7.5, sonicate for 1 hour, and drain.

[0093] The tissue was immersed in a 2% sodium bicarbonate aqueous solution, sonicated for 1 hour, and then drained.

[0094] The tissue was immersed in 500ml of 10% ethanol, sonicated for 1.5h, and then drained.

[0095] The tissue was immersed in 500ml of purified water, sonicated for 2 hours, and drained; the tissue was then freeze-dried to obtain the dried product.

[0096] Take 5g of dried product and enzymatically hydrolyze it with 500ml of pepsin solution (0.1%, pH2) at room temperature for 12 hours. Centrifuge and discard the supernatant. Freeze-dry the precipitate to obtain white flocculent ECM material.

[0097] Example 3

[0098] Take 100g of abdominal wall fascia tissue from a healthy pig, cut it into small pieces, soak it in 500mL of acetone, sonicate it (300W power, 40kHz frequency) for 1 hour, drain it, and rinse it 3 times with 1000mL of purified water.

[0099] The tissue was immersed in 700 mL of the first mixed solution (Triton X-100: acetone: anhydrous ethanol: water = 1.2: 10: 25: 63.8 (mass ratio)), sonicated for 1 hour, and then drained.

[0100] The tissue was immersed in 700 mL of a second mixed solution (Triton X-100: sodium carbonate: anhydrous ethanol: water = 2:3:70:25 (mass ratio)), sonicated for 3 hours, and then drained.

[0101] The tissue was immersed in 700 mL of a third mixed solution (sodium dodecyl sulfate: citric acid: anhydrous ethanol: water = 2.5: 2.5: 25: 70 (mass ratio)), sonicated for 2 hours, and then drained.

[0102] The tissue was immersed in 700 mL of the fourth mixed solution (sodium dodecyl sulfate: acetic acid: anhydrous ethanol: water = 3:3:30:64 (mass ratio)), sonicated for 3 hours, and then drained.

[0103] Immerse the tissue in 500ml of purified water, add PBS solution, adjust the pH to 7.0-7.5, sonicate for 1 hour, and drain.

[0104] The tissue was immersed in a 3% sodium bicarbonate aqueous solution, sonicated for 2 hours, and then drained.

[0105] Immerse the tissue in 500ml of 10% ethanol, sonicate for 2 hours, and drain.

[0106] The tissue was immersed in 500ml of purified water, sonicated for 1 hour, and drained; the tissue was then freeze-dried to obtain the dried product.

[0107] Take 5g of dried product and enzymatically hydrolyze it with 500ml of pepsin solution (0.1%, pH2) at room temperature for 12 hours. Centrifuge and discard the supernatant. Freeze-dry the precipitate to obtain white flocculent ECM material.

[0108] Comparative Example 1

[0109] The preparation steps are the same as in Example 1, except that:

[0110] First mixed solution: Triton X-100: Acetone: Anhydrous Ethanol: Water = 0.6:4:10:85.4

[0111] Second mixed solution: Triton X-100: Sodium carbonate: Anhydrous ethanol: Water = 1:1:60:38

[0112] Third mixed solution: Sodium dodecyl sulfate: Citric acid: Anhydrous ethanol: Water = 1:1:10:88

[0113] Fourth mixed solution: Sodium dodecyl sulfate: Acetic acid: Anhydrous ethanol: Water = 0.8:1:10:88.2.

[0114] Comparative Example 2

[0115] The preparation steps are the same as in Example 1, except that:

[0116] First mixed solution: Triton X-100: Acetone: Anhydrous Ethanol: Water = 1.5:15:30:53.5

[0117] Second mixed solution: Triton X-100: Sodium carbonate: Anhydrous ethanol: Water = 3:4:80:13

[0118] Third mixed solution: Sodium dodecyl sulfate: Citric acid: Anhydrous ethanol: Water = 4:4:40:52

[0119] Fourth mixed solution: Sodium dodecyl sulfate: Acetic acid: Anhydrous ethanol: Water = 3:4:40:53.

[0120] Comparative Example 3

[0121] The preparation steps are the same as in Example 1, except that:

[0122] First mixed solution: Sodium dodecyl sulfate: Citric acid: Anhydrous ethanol: Water = 2.5: 2.5: 25: 70

[0123] Second mixed solution: Sodium dodecyl sulfate: Acetic acid: Anhydrous ethanol: Water = 1.5: 2.5: 25: 71

[0124] Third mixed solution: Triton X-100: Acetone: Anhydrous Ethanol: Water = 1:7:20:72

[0125] Fourth mixed solution: Triton X-100: sodium carbonate: anhydrous ethanol: water = 1.8: 2.5: 72.7: 23.

[0126] Comparative Example 4

[0127] The preparation steps are the same as in Example 1, except that:

[0128] The first mixed solution (Triton X-100: acetone: anhydrous ethanol: water) was not used.

[0129] Comparative Example 5

[0130] The preparation steps are the same as in Example 1, except that:

[0131] No second mixed solution (Triton X-100: sodium carbonate: anhydrous ethanol: water) treatment is performed.

[0132] Comparative Example 6

[0133] The preparation steps are the same as in Example 1, except that:

[0134] The fourth mixed solution (sodium dodecyl sulfate: acetic acid: anhydrous ethanol: water) treatment is not performed.

[0135] Comparative Example 7

[0136] The preparation steps are the same as in Example 1, except that:

[0137] The second mixed solution is: Triton X-100: sodium bicarbonate: anhydrous ethanol: water = 1.8: 2.5: 72.7: 23.

[0138] Experimental Example

[0139] 1. The porcine extracellular matrix from the above examples and comparative examples was subjected to residual detection. The detection method is as follows:

[0140] Cell residue detection: section staining method (H&E staining)

[0141] DNA residue: fluorescence method

[0142] Triton X-100 Residue: High Performance Liquid Chromatography

[0143] Sodium dodecyl sulfate (SDS) residue: Ultraviolet spectrophotometry

[0144] The test results are shown in Tables 1 and 2:

[0145] Table 1 Residual Test Results of Examples

[0146]

[0147] Table 2 Comparative residual test results

[0148]

[0149] 2. Human tolerance tests were conducted on the porcine extracellular matrix from the above examples and comparative examples.

[0150] The test results are shown in Tables 3 and 4.

[0151] Table 3 Results of Human Tolerance Experiments for Examples

[0152]

[0153] Table 4 Results of comparative human tolerance experiments

[0154]

[0155] The comparison between the above embodiments and comparative examples clearly demonstrates that the decellularization method with the specific formulation provided in this application is significantly superior to traditional methods in terms of the thoroughness of cell and DNA removal (cell residue ≤2 vs ≤20, DNA residue ≤5μg / g vs ≤80μg / g). More importantly, the method of this application can greatly reduce the residue of chemical reagents (Triton X-100 residue ≤10μg / g vs ≤100μg / g, SDS residue ≤3mg / g vs ≤10mg / g), thereby achieving a 100% tolerability rate in human trials, while traditional methods all exhibit varying degrees of allergic or irritant reactions. This proves the significant advantages of the method of this application in terms of safety and effectiveness.

[0156] The above description of the embodiments is only for the purpose of helping to understand the method and core ideas of this application. It should be noted that, for those skilled in the art, several improvements and modifications can be made to this application without departing from the principles of this application, and these improvements and modifications also fall within the protection scope of the claims of this application.

Claims

1. A method for removing immunogenicity of porcine-derived extracellular matrix, characterized by, The application relates to a method for preparing a porcine-derived extracellular matrix with immunogenicity removed. The method comprises the following steps: pretreating porcine-derived fascia tissue; subjecting the pretreated porcine-derived fascia tissue to first ultrasonic treatment in a first mixed solution to obtain a first intermediate product; subjecting the first intermediate product to second ultrasonic treatment in a second mixed solution to obtain a second intermediate product; subjecting the second intermediate product to third ultrasonic treatment in a third mixed solution to obtain a third intermediate product; subjecting the third intermediate product to fourth ultrasonic treatment in a fourth mixed solution to obtain a fourth intermediate product; subjecting the fourth intermediate product to fifth ultrasonic treatment in a fifth mixed solution to obtain a fifth intermediate product; subjecting the fifth intermediate product to sixth ultrasonic treatment in a sixth mixed solution to obtain a sixth intermediate product; subjecting the sixth intermediate product to seventh ultrasonic treatment and eighth ultrasonic treatment by sequentially immersing the sixth intermediate product in a first ethanol solution and pure water to obtain a seventh intermediate product; subjecting the seventh intermediate product to biological enzyme hydrolysis to obtain a porcine-derived extracellular matrix with immunogenicity removed. The first mixed solution is prepared by mixing the following raw materials in a certain proportion by weight: 0.8-1.2% triton X-100, 5-10% acetone, 15-25% anhydrous ethanol, and the balance of water. The second mixed solution is prepared by mixing the following raw materials in a certain proportion by weight: 1.5-2.0% triton X-100, 2.0-3.0% sodium carbonate, 70-76.5% anhydrous ethanol, and the balance of water. The third mixed solution is prepared by mixing the following raw materials in a certain proportion by weight: 2.0-3.0% sodium dodecyl sulfate, 2.0-3.0% citric acid, 20-30% anhydrous ethanol, and the balance of water. The fourth mixed solution is prepared by mixing the following raw materials in a certain proportion by weight: 1.0-2.0% sodium dodecyl sulfate, 2.0-3.0% acetic acid, 20-30% anhydrous ethanol, and the balance of water.

2. The method of removing immunogenicity of a porcine-derived extracellular matrix according to claim 1, characterized in that, The fifth mixed solution is a PBS solution with a pH of 7.0-7.5, and the sixth mixed solution is a sodium bicarbonate aqueous solution.

3. The method of removing immunogenicity of a porcine-derived extracellular matrix according to claim 2, characterized in that, The pretreatment comprises the following steps: immersing the porcine-derived fascia tissue in a fat-soluble solvent, subjecting the porcine-derived fascia tissue to ultrasonic treatment for 1-2 hours, draining the solvent, and rinsing the residual solvent with pure water.

4. The method of removing immunogenicity of porcine-derived extracellular matrix according to claim 1, characterized in that, The first ethanol solution is an ethanol solution with a mass fraction of 5-15%. The first ultrasonic treatment is performed for 1-3 hours. The second ultrasonic treatment is performed for 1-3 hours. The third ultrasonic treatment is performed for 1-3 hours. The fourth ultrasonic treatment is performed for 1-3 hours. The fifth ultrasonic treatment is performed for 1-2 hours. The sixth ultrasonic treatment is performed for 1-2 hours.

5. The method of removing immunogenicity of porcine-derived extracellular matrix according to claim 1, characterized in that, The seventh ultrasonic treatment is performed for 1-2 hours. The amount of the first mixed solution, the second mixed solution, the third mixed solution and the fourth mixed solution is not less than 7 times the mass of the pretreated porcine-derived fascia tissue. The amount of the fifth mixed solution is not less than 5 times the mass of the first intermediate product. The amount of the first ethanol solution is not less than 5 times the mass of the second intermediate product. The amount of the pure water is not less than 5 times the mass of the second intermediate product.

6. The method of removing immunogenicity of a porcine-derived extracellular matrix according to claim 5, characterized in that, The mass concentration of the sodium bicarbonate aqueous solution is 2%-4%.

7. The method of removing immunogenicity of porcine-derived extracellular matrix according to claim 2, characterized in that, The fat-soluble solvent is one or more of acetone, n-hexane and ethyl acetate.

8. The method of removing immunogenicity of porcine-derived extracellular matrix according to any one of claims 1 to 7, characterized in that, The biological enzyme includes one or more of trypsin, pepsin and collagenase.

9. A porcine-derived extracellular matrix material prepared by the method of any one of claims 1 to 8, characterized in that, The pig-derived extracellular matrix material has cell residues of ≤2 / visual field, DNA residues of ≤5 μg / g, Triton X-100 residues of ≤10 μg / g and sodium dodecyl sulfate residues of ≤3 mg / g.

10. Application of the pig-derived extracellular matrix material prepared by the method of any one of claims 1-8 to a tissue engineering scaffold, a wound dressing or a biological patch.

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