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Preparation method of natural soft tissue decellularization matrix

A decellularized matrix and soft tissue technology, applied in the fields of biomedicine, regenerative medicine, and clinical medicine, can solve the problems of matrix composition and three-dimensional structure destruction, time-consuming, serious reagent residues, etc., to weaken the interaction, shorten the time, The effect of improving cell removal efficiency

Inactive Publication Date: 2017-10-10
GUANGZHOU SUN SHING BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, the traditional tissue decellularized matrix preparation process easily leads to the destruction of matrix components and three-dimensional structure, and there are serious reagent residues, which may cause severe immune rejection when implanted in tissue
Moreover, the traditional tissue decellularization matrix preparation process takes a lot of time to allow the chemical detergent to penetrate into the tissue, act on the cells, and remove the cellular components in the tissue, so the decellularization efficiency is low

Method used

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  • Preparation method of natural soft tissue decellularization matrix
  • Preparation method of natural soft tissue decellularization matrix
  • Preparation method of natural soft tissue decellularization matrix

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Decellularization of natural soft tissues to obtain acellular matrix, taking the submucosa of the pig small intestine as an example, includes the following steps:

[0062] Material: Take the jejunum section of the pig small intestine and clean it with 0.9wt% physiological saline;

[0063] Pretreatment: Carefully peel and remove the serosal layer and base layer in the intestinal wall of the pig small intestine, scrape off the mucosal layer to obtain the submucosa of the pig small intestine, and clean it with 0.9wt% physiological saline;

[0064] Disinfection: soak in a solution containing 0.02wt% peroxyacetic acid and 5% ethanol for 1h, and rinse with 0.9wt% physiological saline;

[0065] Degreasing: Put the sterilized submucosa of the pig small intestine into a degreasing reagent for 4 hours. The degreasing reagent is chloroform. The total volume of the degreasing reagent is 30 times the volume of the submucosa of the pig small intestine;

[0066] Enzyme treatment: Put the defatt...

Embodiment 5

[0081] With reference to the national standard YY / T 0606.25-2014, the undecellularized submucosa of the pig small intestine and the submucosa of the pig small intestine after the decellularization of Examples 2-1 to 4-3 were tested for the amount of residual DNA. The DNA residue in the submucosa of the pig small intestine treated by the decellularization process is 696.53ng / mg. The test results of Examples 2-1 to 4-3 are shown in Table 4. After the above decellularization process, the submucosal matrix of the pig small intestine The amount of residual DNA in the slab is significantly reduced.

[0082] Table 4 DNA residues in the submucosa of pig small intestine with different process parameters

[0083]

Embodiment 6

[0085] Perform HE staining on the porcine small intestinal submucosal matrix after decellularization in Example 1 and Examples 4-1, 4-2, and 4-3, such as Figure 1A , Figure 1B , Figure 1C , Figure 1D As shown, the staining results show that there are basically no cells in the submucosa of the pig small intestine treated in Example 1 and Examples 4-1, 4-2, and 4-3, and no significant difference is observed under the microscope. It shows that this process can effectively remove the cells in the submucosal matrix of the pig small intestine and reduce the immunogenicity of the matrix.

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Abstract

The invention discloses a preparation method of a natural soft tissue decellularization matrix. The method includes material taking, pretreatment, disinfection, degreasing, enzyme treatment, decellularization, rinsing and sterilization. The method has the advantages of being high in decellularization efficiency, small in damage of matrix components, low in immunogenicity of the decellularization matrix, and good in biocompatibility.

Description

Technical field [0001] The invention relates to the technical fields of clinical medicine, biomedicine and regenerative medicine, in particular to a method for preparing natural soft tissue acellular matrix. Background technique [0002] Tissues are composed of cells and extracellular matrix (ECM). The extracellular matrix is ​​a complex network structure formed by structural proteins and functional proteins, supporting and connecting tissue structures, and regulating the physiological activities of cells (migration, differentiation and proliferation) , To complete a specific function. [0003] Compared with non-animal-derived materials such as metals and plastics, the composition and structure of the cell matrix are closer to human tissues, and the biocompatibility is better, which makes it the most widely used material in tissue engineering repair. However, due to the complex three-dimensional structure of the extracellular matrix, it is difficult to reproduce in vitro extracell...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36
CPCA61L27/3633A61L27/3687A61L2430/40
Inventor 梁丽金杨习锋曾晨光
Owner GUANGZHOU SUN SHING BIOTECH CO LTD
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