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Preparation method of cell substrate protein biological support material used for tissue engineering

A cell matrix and matrix protein technology, applied in medical science, prostheses, etc., can solve problems such as limited sources, destruction of protein natural configuration, and no plasticity of products, and achieve growth promotion, high purity, and good biocompatibility Effect

Inactive Publication Date: 2004-05-05
STOMATOLOGICAL HOSPITAL NO 4 ARMY MEDICAL COLLEGE PLA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has the following disadvantages: ① the composition is single, only collagen; the material is human placenta, the source is limited, and it is not suitable for large-scale use in tissue engineering; ② the final product obtained is liquid, and the product content cannot be detected; ③ the product has no plasticity , cannot be used in the development of tissue engineering products
The disadvantages are: ①The material is treated with protease, which destroys the natural configuration of the protein; ②The material is cross-linked, which destroys the biocompatibility of the protein

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1) After washing the fetal bovine dermis, tendon and bone tissue, they are broken into particles;

[0026] 2) Wash the tissue particles with 0.1M Tris / 1mM EDTA 1:1 mixture for 24 hours, and change the solution once every 6 hours;

[0027] 3) Extract matrix protein from tissue pellets with 0.5M acetic acid buffer for 72 hours, and change the medium once every 24 hours;

[0028] 4) The extract is salted out and precipitated by adding 1M NaCl;

[0029] 5) Dissolve the precipitate with 0.5M acetic acid buffer;

[0030] 6) adding 1M NaCl for salting out and precipitation to obtain the crude matrix protein;

[0031] 7) Dialyze 4 times with distilled water to remove residual acetic acid buffer;

[0032] 8) The crude matrix protein was obtained after freeze-drying, and stored at 4°C after sterilization.

Embodiment 2

[0034] The matrix protein crude product that 6) obtains in embodiment 1 is processed as follows again:

[0035] 1) Dissolve the crude protein with 0.1M Tris buffer (pH 7.0);

[0036] 2) Filter and purify the solution with a 200 mesh filter screen;

[0037] 3) adding different gradients of NaCl to the solution to remove type III collagen to obtain purified matrix protein;

[0038] 4) Dialyze 5 times with distilled water to remove residual acetic acid buffer;

[0039] 5) After freeze-drying, the pure matrix protein (slightly acidic) is obtained, and stored at 4°C after sterilization.

Embodiment 3

[0041] 3) in the embodiment 2, the matrix protein pure product that obtains is processed as follows again:

[0042] 1) dissolve the purified matrix protein with 0.05M acetic acid buffer;

[0043] 2) dialyzing the matrix protein solution by sodium hydroxide solution to make it neutral;

[0044] 3) Distilled water dialysis 6 times to remove residual acetic acid buffer;

[0045] 4) Freeze-drying at -30°C to obtain a matrix protein product, which has a milky white spongy structure;

[0046] 5) After the product is sterilized, store it at 4°C.

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PUM

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Abstract

A cytostromatic protein bioscaffold material for tissue engineering is prepared through using the buffering acetic acid solution to extract the cytostromatic protein, depositing with inorganic salt, and using the inorganic salt with different gradient to remove the III-type collagen from the cytostromatic protein, resulting in purified cytostromatic protein. Its advantages are high stability, purity and adhesion and no toxic by-effect.

Description

technical field [0001] The invention belongs to the technical field of biological material preparation, and relates to a preparation method of cell matrix protein biological material used for tissue engineering. Background technique [0002] Matrix protein is the main protein component of connective tissue. As a scaffold material for tissue engineering, it has excellent mechanical properties. The important reason is that matrix protein can be naturally cross-linked to form the three-dimensional structure required for tissue engineering products. Therefore, matrix proteins can meet the requirements of tissue engineering biomaterials in a wide range. At present, matrix protein components (such as collagen, chondroitin sulfate, and extracellular matrix) are widely used in medicine, such as surgical burn treatment agents, drug carriers, or repair materials (ointment, sutures), etc. However, the application in tissue engineering has higher requirements on matrix protein, and it ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/22A61L27/60
Inventor 金岩聂鑫
Owner STOMATOLOGICAL HOSPITAL NO 4 ARMY MEDICAL COLLEGE PLA
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