Acellular dermal matrix as well as preparation method and application thereof

Abstract

The invention discloses an acellular dermal matrix as well as a preparation method and application thereof, and relates to the technical field of decellularization. The preparation method comprises the following steps: respectively carrying out decellularization treatment on dermis by adopting a decellularization reagent, and then cleaning by adopting a NaCl solution; the decellularization reagent comprises a NaCl hypertonic solution and a NaOH solution; or the decellularization reagent is prepared from trypsin and a Tritonx-100 solution. The method can effectively remove cells, and has the advantages of low DNA content and good mechanical properties.

Description

Technical Field

[0001] The present application relates to the field of decellularization technology, and in particular to a decellularized dermal matrix and a preparation method and application thereof. Background Art

[0002] Acellular dermal matrix is ​​a dermal matrix with low immunogenicity obtained by decellularizing animal dermis using a series of physical and chemical methods to remove cellular components. It is widely used in dermal defects caused by burns and trauma, and is also used for mucosal or soft tissue filling.

[0003] Since the dermal matrix is ​​derived from animal skin tissue, which contains a lot of hair tissue, when the dermal matrix is ​​thick, many hair follicles will remain inside, making it more difficult to remove cells at the hair follicles. The inventor of this application tried to perform hair removal before treatment. However, if the hair is directly plucked, many hairs will break directly and remain in the hair follicles, and it is also very difficult to remove cells at the hair follicles; and if high-temperature water is used for hair removal, the high temperature will destroy the protein structure. Summary of the invention

[0004] The present application provides a decellularized dermal matrix and a preparation method and application thereof, which can effectively remove cells, especially cells at the hair follicles. The decellularized dermal matrix has a lower DNA content and better mechanical properties.

[0005] This application is implemented as follows:

[0006] In a first aspect, the present application provides a method for preparing an acellular dermal matrix, comprising the following steps:

[0007] The dermis was decellularized using a decellularizing agent and then washed with a NaCl solution;

[0008] The decellularization reagent includes a NaCl hypertonic solution and a NaOH solution; or, the decellularization reagent includes a trypsin solution and a Tritonx-100 solution.

[0009] In a possible embodiment, during decellularization, the NaCl hypertonic solution is first used for treatment, and then the NaOH solution is used for treatment; optionally, the concentration of the NaCl hypertonic solution is 0.5-2.0 mol / L, and the concentration of the NaOH solution is 0.1-1.5 mol / L.

[0010] In a possible implementation manner, the treatment time of the NaCl hypertonic solution is 8 to 36 hours, and the treatment time of the NaOH solution is 6 to 36 hours.

[0011] In a possible embodiment, the temperature of the NaCl hypertonic solution during treatment is 30-40°C.

[0012] In a possible embodiment, when performing decellularization treatment, the trypsin solution is first used for treatment, and then the Tritonx-100 solution is used for treatment; optionally, the concentration of the trypsin solution is 0.5 to 5 mg / mL, and the concentration of the Tritonx-100 solution is 3 to 10 mg / mL;

[0013] Optionally, the treatment time of the trypsin solution is 18 to 36 hours, and the treatment time of the Tritonx-100 solution is 18 to 36 hours.

[0014] In a possible implementation manner, in the step of washing with a NaCl solution, the concentration of the NaCl solution is 2 to 4 mol / L.

[0015] In a possible embodiment, the dermis is first disinfected and defatted before being decellularized using a decellularizing agent.

[0016] In a possible embodiment, the disinfection treatment comprises the following steps: firstly treating the dermal matrix with peracetic acid having a concentration of 0.2-0.5%, and then treating the dermal matrix with an ethanol solution having a concentration of 20-30%.

[0017] In a second aspect, the present application provides an acellular dermal matrix, which is prepared by the preparation method of the acellular dermal matrix of the embodiment of the first aspect.

[0018] In a third aspect, the present application provides a use of an acellular dermal matrix prepared by the preparation method of the acellular dermal matrix of the first aspect or the acellular dermal matrix of the second aspect in the preparation of a urethral sling, an artificial skin, or a patch.

[0019] This application has at least the following beneficial effects:

[0020] The preparation method of the decellularized dermal matrix of the present application can rupture the cell membrane by treating the dermal tissue with a NaCl hypertonic solution and a NaOH solution, or treating the dermal tissue with a trypsin solution and a Tritonx-100 solution, and then washing with a NaCl solution, which can effectively precipitate and wash out the DNA, thereby obtaining a decellularized dermal matrix with good decellularization effect and low DNA content. According to the test, the cell removal at the hair follicle of the decellularized dermal matrix is ​​also relatively complete, and the DNA content is less than 6ng / mg. In addition, the tensile strength and elongation at break are high, and it has good mechanical properties. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings required for use in the embodiments will be briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present application and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without paying creative work.

[0022] Figure 1 This is a HE staining image of the acellular dermal matrix of Example 1 of the present application;

[0023] Figure 2 This is a HE staining image of the acellular dermal matrix of Example 2 of the present application;

[0024] Figure 3 This is a HE staining image of the acellular dermal matrix of Example 3 of the present application;

[0025] Figure 4 This is a HE staining image of the acellular dermal matrix of Example 4 of the present application;

[0026] Figure 5 This is a HE staining image of the acellular dermal matrix of Example 5 of the present application;

[0027] Figure 6 This is a HE staining image of the acellular dermal matrix of Example 6 of the present application;

[0028] Figure 7 This is a HE staining image of the acellular dermal matrix of Example 7 of the present application;

[0029] Figure 8 This is a HE staining image of the acellular dermal matrix of Example 8 of the present application;

[0030] Fig. 9 This is a HE staining image of the acellular dermal matrix of Example 10 of the present application;

[0031] Fig.10 This is a HE staining image of the acellular dermal matrix of Comparative Example 1 of the present application;

[0032] Fig.11 This is a HE staining image of the acellular dermal matrix of Comparative Example 2 of the present application;

[0033] Fig.12 This is a HE staining image of the acellular dermal matrix of Comparative Example 3 of the present application;

[0034] Fig.13 This is the HE staining image of the acellular dermal matrix of Comparative Example 4 of the present application. DETAILED DESCRIPTION

[0035] The embodiments of the present application will be described in detail below in conjunction with the examples, but it will be appreciated by those skilled in the art that the following examples are only used to illustrate the present application and should not be considered as limiting the scope of the present application. In the examples, if specific conditions are not specified, they are carried out according to normal conditions or the conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be purchased commercially.

[0036] The following is a detailed description of the acellular dermal matrix and its preparation method and application according to the embodiment of the present application:

[0037] In a first aspect, the present invention provides an acellular dermal matrix and a preparation method thereof, which comprises the following steps:

[0038] (1) After the skin is taken from the animal tissue, the hair is shaved first, and then a skin removal machine is used to remove 2 to 8 cm of skin tissue. The removed dermis is disinfected and degreased.

[0039] The disinfection treatment can clean away viruses and pathogens in the dermis. Optionally, the disinfection treatment includes the following steps: firstly treating the dermis matrix with 0.2-0.5% peracetic acid, and then treating the dermis matrix with 20-30% ethanol solution.

[0040] Peracetic acid can kill some bacteria and fungi such as Escherichia coli, Staphylococcus aureus, Candida albicans, Staphylococcus aureus, etc. Optionally, the concentration of peracetic acid is 0.2%, 0.3%, 0.4% or 0.5%. After disinfection with peracetic acid, 20-30% ethanol solution is used to disinfect the dermal matrix, so that the disinfection effect is better. Optionally, the concentration of the ethanol solution is any one of 20%, 25% and 30%, or a value between any two.

[0041] (2) Degreasing the disinfected dermal matrix; wherein the degreasing can be firstly treated with dichloromethane and then with anhydrous ethanol, so that the fat removal rate is greater than 95% or more.

[0042] (3) Decellularizing the dermis with a decellularizing agent, and then washing with a NaCl solution; the decellularizing agent includes a NaCl hypertonic solution and a NaOH solution; or, the decellularizing agent includes trypsin and a Tritonx-100 solution.

[0043] In some embodiments, during the decellularization treatment, a NaCl hypertonic solution is first used for treatment, and then a NaOH solution is used for treatment.

[0044] The dermal matrix is ​​treated with a NaCl hypertonic solution, which has a swelling effect on cells, causing them to burst and release nucleic acid substances in the cells. The role of NaOH is to transform the cell membrane from a lipid bilayer structure to a vesicle, thereby allowing the cells to be more fully lysed and the cell contents to be released. NaOH reacts with DNA to unwind the double-stranded structure, which is conducive to the subsequent elution of DNA with NaCl solution. Moreover, according to detection, it was found that the cells at the hair follicles can be fully removed by first treating with a NaCl hypertonic solution and then with a NaOH solution.

[0045] Exemplarily, the concentration of the NaCl hypertonic solution is 0.5-2.0 mol / L, for example, any one of 0.5 mol / L, 1.0 mol / L, 1.5 mol / L and 2.0 mol / L or a value between any two thereof. Exemplarily, the concentration of the NaOH solution is 0.1-1.5 mol / L, for example, any one of 0.1 mol / L, 0.3 mol / L, 0.5 mol / L, 0.7 mol / L, 1.0 mol / L, 1.2 mol / L and 1.5 mol / L or a value between any two thereof.

[0046] The treatment time of the NaCl hypertonic solution is 8 to 36 hours, for example, any one of 8 hours, 12 hours, 18 hours, 24 hours, 28 hours, 32 hours and 36 hours, or a value between any two of them. If the treatment time of the NaCl hypertonic solution is too short, the cells will not swell sufficiently, which will result in insufficient release of nucleic acid substances in the cells.

[0047] Furthermore, the temperature during the NaCl hypertonic solution treatment is 30-40°C.

[0048] The applicant has found that when the temperature during the treatment with the NaCl hypertonic solution is 30-40°C, the decellularization effect is better than when the temperature is lower than 30°C (e.g., 25°C). Optionally, the temperature during the treatment with the NaCl hypertonic solution is any one of 30°C, 32°C, 35°C, 37°C and 40°C, or a value between any two of them.

[0049] The treatment time of the NaOH solution is 6 to 36 hours, for example, any one of 6 hours, 12 hours, 18 hours, 24 hours, 28 hours, 32 hours and 36 hours, or a value between any two of them. If the treatment time of the NaOH solution is too short, it may lead to insufficient cell lysis, and the double-stranded structure of DNA cannot be fully unwound, resulting in a high amount of residual DNA.

[0050] In another embodiment, when performing decellularization, trypsin solution is first used for treatment, and then Tritonx-100 solution is used for treatment; optionally, the concentration of trypsin solution is 0.5-5 mg / mL, and the concentration of Tritonx-100 solution is 3-10 mg / mL. It should be noted that the trypsin solution can be provided by a pancreatic enzyme solution, and the pancreatic enzyme contains trypsin, and optionally, also contains pancreatic amylase and pancreatic lipase.

[0051] Trypsin is used to process dermal matrix, and relies on the cleavage of the peptide bond of the carboxyl side carbon of arginine and lysine, thereby causing the cell components to be separated from the dermal matrix. Optionally, the concentration of trypsin is 0.5mg / mL, 1mg / mL, 2mg / mL, 2.5mg / mL, 3mg / mL, 4mg / mL and 5mg / mL, any one or any two values ​​between them. Optionally, the concentration of Tritonx-100 solution is 3mg / mL, 4mg / mL, 5mg / mL, 6mg / mL, 7mg / mL, 8mg / mL, 9mg / mL and 10mg / mL, any one or any two values ​​between them.

[0052] (4) After decellularization, the cells were washed with NaCl solution.

[0053] NaCl solution can precipitate DNA, and the use of NaCl solution can easily wash and rinse out DNA, especially the unwound DNA. Optionally, in the step of using NaCl solution for washing, the concentration of the NaCl solution is 2 to 4 mol / L, for example, any one of 2 mol / L, 2.5 mol / L, 3 mol / L, 3.5 mol / L and 4 mol / L, or a value between any two of them.

[0054] In a second aspect, the present application provides an acellular dermal matrix, which is prepared by the preparation method of the acellular dermal matrix of the first aspect.

[0055] The acellular dermal matrix prepared by the preparation method of the acellular dermal matrix of the embodiment of the present application has completely removed cells and a DNA content of less than 6 ng / mg. In addition, the tensile strength and elongation at break are high, and the acellular dermal matrix has good mechanical properties.

[0056] In a third aspect, the present application provides a use of an acellular dermal matrix prepared by the preparation method of the acellular dermal matrix of the first aspect or the acellular dermal matrix of the second aspect in the preparation of a urethral sling, an artificial skin, or a patch.

[0057] The acellular dermal matrix prepared in the present application has completely removed cells, low DNA content, high biocompatibility, and low immunogenicity, and can be used for the preparation of medical devices such as patches, artificial skin, and urethral slings.

[0058] The acellular dermal matrix of the present application and its preparation method and application are further described in detail below in conjunction with the examples.

[0059] Example 1

[0060] This embodiment provides a method for preparing an acellular dermal matrix, which comprises the following steps:

[0061] (1) After the skin is taken from the animal tissue, the hair is shaved first, and then a 6 cm thick skin tissue is taken using a skin-taking machine, and the removed dermis is disinfected and degreased. Among them, the disinfection treatment first uses a 0.5% peracetic acid concentration to treat the dermal matrix, and then uses a 25% ethanol solution to treat the dermal matrix. The degreasing treatment is carried out using a mixed solution of dichloromethane and anhydrous ethanol in a volume ratio of 2:1.

[0062] (2) The dermal matrix after disinfection and degreasing was first treated with a 1 mol / L NaCl hypertonic solution at 37°C for 24 h, and then treated with a 0.5 mol / L NaOH solution for 24 h.

[0063] (3) Use a 2 mol / L NaCl solution for cleaning for 2 h.

[0064] Example 2

[0065] This embodiment provides a method for preparing an acellular dermal matrix. Compared with Embodiment 1, the only difference is that the treatment time of the NaOH solution is 18 hours.

[0066] Example 3

[0067] This embodiment provides a method for preparing an acellular dermal matrix. Compared with Embodiment 1, the only difference is that the treatment time of the NaOH solution is 9 hours.

[0068] Example 4

[0069] This embodiment provides a method for preparing an acellular dermal matrix. Compared with Embodiment 1, the only difference is that the treatment time of the NaOH solution is 6 hours.

[0070] Example 5

[0071] This embodiment provides a method for preparing an acellular dermal matrix. Compared with Embodiment 1, the only difference is that the treatment time of the NaOH solution is 3 hours.

[0072] Example 6

[0073] This embodiment provides a method for preparing an acellular dermal matrix. Compared with Embodiment 2, the temperature condition for the treatment with the NaCl hypertonic solution is 25°C.

[0074] Example 7

[0075] This embodiment provides a method for preparing acellular dermal matrix. Compared with Example 2, the only difference is that in step (2) of this embodiment, a trypsin solution with a concentration of 2.5 mg / mL is first used for treatment for 24 hours, and then a Tritonx-100 solution with a concentration of 5 mg / mL is used for treatment for 24 hours.

[0076] Example 8

[0077] This embodiment provides a method for preparing an acellular dermal matrix. Compared with Example 7, the only difference is that the concentration of the trypsin solution is 5 mg / mL and the concentration of the Tritonx-100 solution is 3 mg / mL.

[0078] Example 9

[0079] This embodiment provides a method for preparing an acellular dermal matrix. Compared with Example 7, the only difference is that the concentration of the trypsin solution in Example 9 is 2 mg / mL and the treatment time is 6 h.

[0080] Example 10

[0081] This embodiment provides a method for preparing an acellular dermal matrix. Compared with Embodiment 1, the only difference is that the concentration of the NaCl hypertonic solution is 0.5 mol / L, and the concentration of the NaOH solution is 1.0 mol / L.

[0082] Embodiment 11

[0083] This embodiment provides a method for preparing an acellular dermal matrix. Compared with Embodiment 7, the difference is that this embodiment uses pancreatic enzyme, wherein the pancreatic enzyme contains trypsin, and the concentration of trypsin is also 2.5 mg / mL.

[0084] Comparative Example 1

[0085] This comparative example provides a method for preparing an acellular dermal matrix. Compared with Example 2, the only difference is that the step of treating with a NaCl hypertonic solution in step (2) of Example 3 is omitted.

[0086] Comparative Example 2

[0087] This comparative example provides a method for preparing an acellular dermal matrix. Compared with Example 2, the only difference is that step (2) is to treat with sodium dodecyl sulfate (SDS) at a concentration of 10 mg / mL for 3 hours.

[0088] Comparative Example 3

[0089] This comparative example provides a method for preparing an acellular dermal matrix. Compared with Example 2, the only difference is that step (2) is to treat with sodium dodecyl sulfate (SDS) at a concentration of 20 mg / mL for 3 hours.

[0090] Comparative Example 4

[0091] This comparative example provides a method for preparing an acellular dermal matrix. Compared with Example 1, the only difference is that in step (2), trypsin at a concentration of 2.5 mg / mL is used for treatment for 24 hours, and then sodium dodecyl sulfate at a concentration of 10 mg / mL is used for treatment for 30 minutes.

[0092] Comparative Example 5

[0093] This comparative example provides a method for preparing an acellular dermal matrix. Compared with Example 2, the only difference is that the step (3) of washing with a NaCl solution in Example 2 is omitted.

[0094] Comparative Example 6

[0095] This comparative example provides a method for preparing an acellular dermal matrix. Compared with Example 7, the only difference is that the step (3) of washing with a NaCl solution in Example 7 is omitted.

[0096] Comparative Example 7

[0097] This comparative example provides a method for preparing an acellular dermal matrix. Compared with Example 7, the only difference is that step (2) is to first treat with a 5 mg / mL Tritonx-100 solution for 24 hours, and then treat with a 5 U / mL DNA enzyme for 24 hours.

[0098] Test Example 1

[0099] The acellular dermal matrices prepared in Examples 1 to 8, Example 10 and Comparative Examples 1 to 4 were sectioned and HE stained. The HE staining images are shown in FIG. Figures 1 to 13 The circled part in the figure is the remaining cell nucleus component.

[0100] Comparative Example 2 ( Figure 2 ) and Comparative Example 1 ( Fig.10), compared with Example 2, Comparative Example 1 omitted the step of treating the dermal matrix with a NaCl hypertonic solution. The HE staining of the decellularized dermal matrix of Comparative Example 1 showed that there were still cellular components, and the cells at the hair follicles were obviously residual, indicating that omitting the NaCl hypertonic solution treatment would lead to incomplete decellularization. In addition, the HE staining of the decellularized dermal matrix of Example 10 also showed that the cells were completely removed, indicating that the concentration of the NaCl hypertonic solution and the concentration of the NaOH solution in the examples of the present application can effectively remove the cellular components of the dermal matrix, and the cells at the hair follicles can also be completely removed.

[0101] Comparative Example 2 ( Figure 2 )、Example 7( Figure 7 ) and Comparative Examples 2 to 4 ( Figure 11 to Figure 13 ), the HE staining results of the decellularized dermal matrices of Comparative Examples 2 to 4 all showed that there were still residual cell components, while the cell components of Example 2 and Example 7 were completely removed, indicating that the selection of reagents during decellularization has an important influence on the decellularization process. Decellularization using SDS or trypsin combined with SDS could not achieve the effect of complete cell removal. Example 2 used NaCl hypertonic solution and NaOH solution for treatment in sequence, and Example 7 used trypsin solution and Tritonx-100 solution for treatment, both of which could completely remove the cells of the dermal matrix. Moreover, Figure 2 As shown, the cells at the hair follicles of Example 2 were completely removed. In addition, the HE staining of the decellularized dermal matrix of Example 8 showed that the cells were completely removed, indicating that the concentrations of the trypsin solution and Tritonx-100 solution of the examples of the present application can effectively remove the cell components of the dermal matrix.

[0102] By comparing the HE staining results of Examples 1 to 5, it can be found that the HE staining images of the decellularized dermal matrices of Examples 4 and 5 show that there are still a few cell components remaining, while the cell components of the decellularized dermal matrices of Examples 1 to 3 are completely removed, indicating that when the dermis is decellularized using a NaCl hypertonic solution and a NaOH solution, the treatment time has an important influence on the decellularization effect. Only when the NaOH solution treatment time is 9 hours or more, a dermal matrix with completely removed cells can be obtained.

[0103] The treatment condition of the NaCl hypertonic solution in Example 2 is 37°C, while the treatment condition of the NaCl hypertonic solution in Example 6 is 25°C. By comparing the HE staining results of Example 2 and Example 6, it can be found that the cells of the decellularized dermal matrix in Example 2 are completely removed, while there are still a few cells remaining in the decellularized dermal matrix in Example 6, indicating that the treatment temperature of the NaCl hypertonic solution at 37°C is more conducive to complete cell removal.

[0104] Test Example 2

[0105] The DNA content of the acellular dermal matrix prepared by the preparation method of the acellular dermal matrix of Example 1, Example 2, Example 7, Example 9 and Comparative Examples 5 to 6 was detected, and the detection referred to YYT 1876-2023 (Determination of DNA Residue in Animal-Derived Biomaterials for Tissue Engineering Medical Products: Fluorescence Staining Method), and the results are recorded in Table 1.

[0106] Table 1. Residual DNA content in acellular dermal matrix

[0107] DNA residue / (ng / mg) Example 1 3.11 Example 2 5.15 Example 7 5.17 Example 9 5.78 Comparative Example 5 12.85 Comparative Example 6 19.47

[0108] Referring to the results in Table 1, by comparing the results of Example 1, Example 2 and Comparative Example 5, it can be found that the residual DNA content of the acellular dermal matrix of Example 1 and Example 2 is less than 6 ng / mg, while the residual DNA content of the acellular dermal matrix of Comparative Example 5, which omits the step of washing with NaCl solution, is greater than 10 ng / mg, indicating that omitting the step of washing with NaCl solution will lead to a significant increase in the residual DNA of the acellular matrix. Moreover, by comparing Example 1 and Example 2, it can be found that the residual DNA content of the acellular dermal matrix of Example 1, which has a longer NaOH solution treatment time, is lower, indicating that extending the NaOH solution treatment time is more conducive to reducing the residual DNA content.

[0109] By comparing the results of Example 7 and Comparative Example 6, it can be found that the residual DNA content of the decellularized matrix of Example 7 is less than 6 ng / mg, while the residual DNA content of the decellularized dermal matrix of Comparative Example 6 in which the NaCl solution washing step is omitted is greater than 19 ng / mg, indicating that omitting the NaCl solution washing step will lead to a significant increase in the residual DNA of the decellularized matrix.

[0110] Test Example 3

[0111] The acellular dermal matrices prepared in Example 2, Example 7 and Comparative Example 7 were cut into dumbbell shapes with a size of 1 cm*5 cm, and mechanical properties were tested with reference to (GB / T 1040.3-2006 Determination of tensile properties of plastics). The measured average tensile strength and average elongation at break are shown in Table 2.

[0112] Table 2. Mechanical testing results of acellular dermal matrix

[0113] Tensile strength / MPa Elongation at break Example 2 4.11 38.23％ Example 7 3.59 48.17％ Comparative Example 7 3.45 29.12％

[0114] From the results in Table 2, it can be seen that the tensile strength and elongation at break of Example 2 and Example 7 are higher than those of Comparative Example 7, especially the elongation at break, which is more than 9% higher than that of Comparative Example 7. This indicates that the decellularization process using trypsin combined with Tritonx-100 is easier to obtain a decellularized dermal matrix with high tensile strength and good elongation at break than the decellularization process using Tritonx-100 combined with DNA protease.

[0115] The above description is only a specific embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A method for preparing an acellular dermal matrix, It is characterized in that The following steps are involved: The dermis was decellularized using a decellularizing agent and then washed with a NaCl solution; The decellularization reagent includes a NaCl hypertonic solution and a NaOH solution; or, the decellularization reagent includes a trypsin solution and a Tritonx-100 solution.

2. The method for preparing the acellular dermal matrix according to claim 1, It is characterized in that When performing decellularization treatment, the NaCl hypertonic solution is first used for treatment, and then the NaOH solution is used for treatment; optionally, the concentration of the NaCl hypertonic solution is 0.5-2.0 mol / L, and the concentration of the NaOH solution is 0.1-1.5 mol / L.

3. The method for preparing the acellular dermal matrix according to claim 2, It is characterized in that The treatment time of the NaCl hypertonic solution is 8 to 36 hours, and the treatment time of the NaOH solution is 6 to 36 hours.

4. The method for preparing the acellular dermal matrix according to claim 2, It is characterized in that The temperature during the treatment with the NaCl hypertonic solution is 30-40°C.

5. The method for preparing the acellular dermal matrix according to claim 1, It is characterized in that When performing decellularization, the trypsin solution is first used for treatment, and then the Tritonx-100 solution is used for treatment; optionally, the concentration of the trypsin solution is 0.5-5 mg / mL, and the concentration of the Tritonx-100 solution is 3-10 mg / mL; Optionally, the treatment time of the trypsin solution is 18 to 36 hours, and the treatment time of the Tritonx-100 solution is 18 to 36 hours.

6. The method for preparing the acellular dermal matrix according to any one of claims 1 to 5, It is characterized in that In the step of using NaCl solution for cleaning, the concentration of the NaCl solution is 2-4 mol / L.

7. The method for preparing the acellular dermal matrix according to any one of claims 1 to 5, It is characterized in that Before decellularizing the dermis using a decellularizing agent, the dermis is first disinfected and defatted.

8. The method for preparing the acellular dermal matrix according to claim 7, It is characterized in that The disinfection treatment comprises the following steps: firstly treating the dermal matrix with peracetic acid having a concentration of 0.2-0.5%, and then treating the dermal matrix with an ethanol solution having a concentration of 20-30%.

9. An acellular dermal matrix, It is characterized in that The acellular dermal matrix is ​​prepared by the preparation method of any one of claims 1 to 8.

10. Use of the acellular dermal matrix according to claim 9 or the acellular dermal matrix prepared by the preparation method of the acellular dermal matrix according to any one of claims 1 to 8 in the preparation of a urethral sling, artificial skin, or patch.

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