Cow achilles tendon collagen purification method and sponge preparation thereof

A technology of tendon collagen and collagen, applied in the field of collagen sponge preparation, can solve the problems of collagen sponge cracks, high market price, complicated process, etc., and achieve the effects of low production cost, wide promotion value, and simple production process

Active Publication Date: 2016-05-25
浙江崇山生物制品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] On the other hand, if the degree of cross-linking of the collagen solution is not well controlled, cracks will appear in the collagen sponge during the removal of the cross-linking agent, and it is difficult to remove the cross-linking agent. However, it is difficult to completely remove glutaraldehyde from the highly hydrophilic collagen sponge, and it needs to be washed repeatedly until there is no glutaraldehyde. This process is not only complicated, but also difficult to completely remove glutaraldehyde. Harmful to surrounding cells, seriously affecting biocompatibility, resulting in lower pass rate of collagen sponge and high market price

Method used

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  • Cow achilles tendon collagen purification method and sponge preparation thereof
  • Cow achilles tendon collagen purification method and sponge preparation thereof
  • Cow achilles tendon collagen purification method and sponge preparation thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Embodiment 1 Purification of collagen and the specific steps of sponge preparation thereof are as follows:

[0014] 1) Use a blade to remove impurities such as fascia, muscle and fat from the bovine Achilles tendon that has been frozen at -20°C, cut into 0.3mm thick with a slicer, and then fully pulverize with a grinder;

[0015] 2) Remove impurity protein with dilute alkali solution for 6-8 hours, wash with distilled water until neutral, then degrease with alcohol, wash with distilled water until neutral and no peculiar smell;

[0016] 3) Dissolved in a dilute acid solution with a pH of 2 to 3;

[0017] 4) adding an appropriate amount of pepsin to the above solution, the weight ratio of collagen to pepsin is 25:1, stirring at 4°C for 48 hours;

[0018] 5) Centrifuge the extract, collect the supernatant, adjust the collagen solution to neutral with 10mol / L sodium hydroxide, collect the obtained collagen precipitate, first add 0.5mol / L acetic acid to dissolve the collag...

Embodiment 2

[0021] Embodiment 2 The specific steps for the preparation of purified collagen and its sponge are as follows:

[0022] 1) Use a blade to remove impurities such as fascia, muscle and fat from the bovine Achilles tendon that has been frozen at -20°C, cut into 0.3mm thick with a slicer, and then fully pulverize with a grinder;

[0023] 2) Remove impurity protein with dilute alkali solution for 6-8 hours, wash with distilled water until neutral, then degrease with alcohol, wash with distilled water until neutral and no peculiar smell;

[0024] 3) Dissolved in a dilute acid solution with a pH of 2 to 3;

[0025] 4) adding an appropriate amount of pepsin to the above solution, the weight ratio of collagen to pepsin is 50:1, stirring at 4°C for 60 hours;

[0026] 5) Centrifuge the extract, collect the supernatant, adjust the collagen solution to neutral with 10mol / L sodium hydroxide, collect the obtained collagen precipitate, first add 0.5mol / L acetic acid to dissolve the collagen ...

Embodiment 3

[0029] Embodiment 3 The specific steps for the preparation of purified collagen and its sponge are as follows:

[0030] 1) Use a blade to remove impurities such as fascia, muscle and fat from the bovine Achilles tendon that has been frozen at -20°C, cut into 0.3mm thick with a slicer, and then fully pulverize with a grinder;

[0031] 2) Remove impurity protein with dilute alkali solution for 6-8 hours, wash with distilled water until neutral, then degrease with alcohol, wash with distilled water until neutral and no peculiar smell;

[0032] 3) Dissolved in a dilute acid solution with a pH of 2 to 3;

[0033] 4) adding an appropriate amount of pepsin to the above solution, the weight ratio of collagen to pepsin is 100:1, stirring at 4°C for 72 hours;

[0034] 5) Centrifuge the extract, collect the supernatant, adjust the collagen solution to neutral with 10mol / L sodium hydroxide, collect the obtained collagen precipitate, first add 0.5mol / L acetic acid to dissolve the collagen...

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Abstract

The present invention relates to a collagen purification method. The purpose of the present invention is that cow achilles tendon is completely crushed by using a new method, a pH value adjusting and collagen precipitation method is used to replace salt precipitation and dialysis desalination during collagen purification, a chemical cross-linking agent is not added during a sponge molding process, the process is simple, the collagen extraction rate is improved, the high qualification rate collagen sponge is achieved, and bleeding stopping sponge is prepared by using the collagen.

Description

technical field [0001] The invention relates to a preparation method of a collagen sponge with a high qualified rate achieved by a simple process. Background technique [0002] Collagen is the most abundant and widely distributed protein in vertebrates. Fibrous collagen forms the structural basis of connective tissue in all mammals, and is also the main component of many tissues, especially connective tissue, accounting for about 25% of the total protein in the body. The unique triple-helix structure of collagen endows it with good biocompatibility, biodegradability, and non-toxicity, which makes it show great potential in medicine and biological tissue engineering. [0003] However, the collagen sponges on the market are expensive, have poor flexibility, and are cross-linked with potentially toxic cross-linking agents, which will seriously affect the biocompatibility of the sponge. The general collagen sponge process is complicated, and the process mainly includes raw mat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/78C07K1/30A61L15/32
Inventor 彭新生
Owner 浙江崇山生物制品有限公司
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