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Method for preparing decellularization cartilage matrix material

A decellularized cartilage and matrix material technology, applied in the field of biomaterial preparation, can solve problems such as long soaking time, failure to use the product, and high concentration of lye

Pending Publication Date: 2018-10-16
BEIJING QINGYUAN WEIYE BIO TISSUE ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The extracellular matrix made by the method described in the existing patent CN1324567A has a high concentration of lye, a long soaking time, and a high processing temperature, which will cause fiber digestion, collapse and void layers, which will cause cells to fail to grow into the product when the product is applied. , affecting the use of the final product
The lye concentration is low, the soaking time is short, and the treatment temperature is low, which will cause the cells to be unclean, cause the immune reaction of the matrix, and cause the product to fail to use

Method used

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  • Method for preparing decellularization cartilage matrix material
  • Method for preparing decellularization cartilage matrix material
  • Method for preparing decellularization cartilage matrix material

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0041] The invention provides a preparation method of decellularized cartilage matrix material. In all embodiments of the present invention, all possess following common features: described preparation method comprises the following steps:

[0042] (1) placing human or animal cartilage tissue in a fixative for fixation;

[0043] (2) Wash the fixed cartilage tissue with a suitable solution, and the number of times of washing can be several times;

[0044] (3) placing the washed cartilage tissue in an alkaline solution for decellularization;

[0045] (4) extracting and cleaning the decellularized cartilage tissue;

[0046] The fixing agent is glutaraldehyde, formaldehyde or peracetic acid;

[0047] The suitable solution may be distilled water;

[0048] The alkaline solution is selected from NaOH or KOH solutions with a concentration ranging from 4% to 15%. Using this concentration of alkali solution can remove the cells of the treated decellularized cartilage matrix material w...

experiment example 1

[0061] Experimental example 1, concrete operation of the present invention

[0062] Cartilage obtained from human or animal tissues is removed, fat is removed, and fixed in a fixative. The fixative can be glutaraldehyde, formaldehyde or peracetic acid. The tissue is left in fixative for at least 2 hours, or until the tissue is fixed. After the tissue is fixed, it is washed with a suitable solution, the solution may be distilled water, and the number of times of washing may be several times.

[0063] The tissue is then decellularized in an alkaline solution, which may be NaOH or KOH solution. The standing time is 15-30 hours, the temperature is 15-35°C, and the concentration of lye is 4%-15%.

[0064] After the tissue is decellularized, it is extracted and cleaned with physiological saline, and the soaking time is 5-7 hours to ensure the biological safety of the tissue. In practice, it has been found that if there is no immersion cleaning, the cytotoxicity of the tissue is n...

experiment example 2

[0066] Experimental example 2, the performance effect verification of the cartilage matrix material prepared by the method of the present invention

[0067] The cartilage matrix material obtained by the preparation method of the present invention is subjected to conventional detection methods in the field, such as the detection method described in the invention patent application 201711056849.0, to detect the following performance indicators, and obtain the following results:

[0068] The cytotoxicity test result of the cartilage matrix material obtained by the preparation method of the present invention: good cytocompatibility;

[0069]

[0070] Detection of immunogenic components: no immunogen, the number of residual cells is 0;

[0071] Decellularization effect data: DNA residue figure 1 Shown, cell residue: 0;

[0072] Such as figure 2 As shown, the porosity data of the acellular cartilage matrix material obtained by the preparation method of the present invention ob...

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Abstract

The invention discloses a 'method for preparing a decellularization cartilage matrix material', and belongs to the technical field of biological material preparation. The method comprises the following steps: (1) carrying out immobilization treatment on human or animal cartilage tissue in an immobilization agent; (2) washing the cartilage tissue after the immobilization treatment by using an appreciate solution, wherein the washing time can be more times; (3) carrying out decellularization treatment on the washed cartilage tissue in an alkali solution; (4) carrying out soaking washing treatment on the cartilage tissue after the decellularization treatment, wherein the immobilization agent is glutaraldehyde, formaldehyde or peracetic acid, the appropriate solution may be distilled water, and the alkali solution is selected from 4-15% NaOH or KOH solutions, and a washing liquid used in soaking washing treatment is normal saline. The decellularization cartilage matrix material prepared byusing the method has a porosity of 70-90% and a pore size of 12-67mu m.

Description

technical field [0001] The invention belongs to the technical field of biological material preparation, and in particular relates to a preparation method of an acellular cartilage matrix material. Background technique [0002] "Cartilage" is specialized dense connective tissue. It is composed of a large number of interstitial cells and encapsulated chondrocytes. There are at least a few cells in the cartilage capsule, and the interstitial cells are firm and elastic. Animals have cartilage in the outer ear, trachea wall, between the vertebral bones, and at the end of the parabone. "Cartilage" is a type of connective tissue in a human or vertebrate body. During the embryonic period, most of the human skeleton is composed of cartilage. Adults only have cartilage on the tip of the nose, the outer ear, the tip of the ribs, and the connecting surfaces of the vertebrae. "Cartilage" can be divided into hyaline cartilage, elastic cartilage and fibrocartilage. It is a kind of sli...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61L27/54
CPCA61L27/3612A61L27/3633A61L27/3654A61L27/3687A61L27/54A61L2430/06A61L2430/40
Inventor 胡杰李燕青曹志华
Owner BEIJING QINGYUAN WEIYE BIO TISSUE ENG
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