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Acellular vascular matrix material preparation method

A matrix material and decellularization technology, applied in the field of biomaterial preparation, can solve the problems of short soaking time, fiber digestion, long soaking time, etc.

Pending Publication Date: 2018-09-14
BEIJING QINGYUAN WEIYE BIO TISSUE ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The extracellular matrix made by the method described in the existing patent CN1284265A has a high concentration of lye, a long soaking time, and a high processing temperature, resulting in 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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  • Acellular vascular matrix material preparation method
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Examples

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preparation example Construction

[0035] The invention provides a preparation method of decellularized vascular matrix material. In all embodiments of the present invention, all possess following common features: described preparation method comprises the following steps:

[0036] (1) placing human or animal vascular tissue in a fixative for fixation;

[0037] (2) placing the fixed vascular tissue in an alkaline solution for decellularization;

[0038] (3) extracting and cleaning the decellularized vascular tissue;

[0039] The fixing agent is glutaraldehyde, formaldehyde or peracetic acid;

[0040] The alkaline solution is selected from NaOH or KOH solutions with a concentration ranging from 4% to 15%;

[0041] The use of this lye concentration can remove the cells of the processed decellularized vascular matrix material without causing the body's immune response, and at the same time allow the processed decellularized vascular matrix material to have sufficient gaps. The porosity of the decellularized va...

experiment example 1

[0055] Experimental example 1, concrete operation of the present invention

[0056] Blood vessels obtained from human or animal tissues are removed, fat and fur are removed, and fixed in a fixative, which 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.

[0057] 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%.

[0058] 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 tiss...

experiment example 2

[0060] Experimental example 2, performance effect verification of the vascular matrix material prepared by the method of the present invention

[0061] The vascular 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:

[0062] Cytotoxicity test results: the cells are non-toxic; the detection of immunogenic components is 0;

[0063] Decellularization effect data: such as figure 2 As shown, the amount of residual cells was 0.

[0064] The void data of the decellularized vascular matrix material prepared by the method of the present invention is obtained through electron microscope observation, counting and statistics: the void ratio is 70%-90%, and the void can reach 19 μm-79 μm.

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Abstract

The invention belongs to the technical field of biological material preparation and discloses an acellular vascular matrix material preparation method. The preparation method includes steps: (1) subjecting human or animal vascular tissues to fixing treatment in a fixing agent; (2) subjecting the fixed vascular tissues to decellularization treatment in alkaline solution; (3) subjecting the vasculartissues to extraction and cleaning treatment after decellularization treatment, wherein the fixing agent refers to glutaraldehyde, formaldehyde or peracetic acid, the alkaline solution is selected from NaOH or KOH solution in concentration range of 4%-15%, and cleaning liquid used in extraction and cleaning treatment is normal saline. Porosity of an acellular vascular matrix material prepared according to the preparation method is 70%-90%, voids reach 19-79micron, cell ingrowth can be realized in product application, and collapse and gap layers due to large voids caused by fiber digestion areavoided.

Description

technical field [0001] The invention belongs to the technical field of biological material preparation, and in particular relates to a preparation method of decellularized vascular matrix material. Background technique [0002] Components of the extracellular matrix are often conserved across species and can be tolerated by xenogeneic receptors. Extracellular matrices obtained from different tissues, including skin, heart valves, blood vessels, nerves, skeletal muscles, tendons, ligaments, small intestinal submucosa, have been extensively studied for tissue engineering and regenerative material applications. The goal of decellularization is to efficiently remove all cellular and nuclear material while minimizing adverse effects on the bioactivity and mechanical integrity of the material. [0003] The extracellular matrix made by the method described in the existing patent CN1284265A has a high concentration of lye, a long soaking time, and a high processing temperature, res...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/50A61L27/56
CPCA61L27/3633A61L27/3687A61L27/50A61L27/507A61L27/56A61L2430/40
Inventor 胡杰李燕青曹志华
Owner BEIJING QINGYUAN WEIYE BIO TISSUE ENG
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