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Extraction method of bovine tendon type I collagen

An extraction method, the technology of bovine Achilles tendon, applied in chemical instruments and methods, animal/human protein, connective tissue peptide, etc., can solve the problems of encapsulation reaction, high price, and dependence on imports, and achieve good biocompatibility and equipment The effect of low requirements and simple preparation method

Active Publication Date: 2017-10-03
QIANFOSHAN HOSPITAL OF SHANDONG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The currently extracted materials may cause rejection, and due to the reaction of local tissues, it may cause stimulation to the brain tissue, forming scars or wrapping reactions, meningitis symptoms or bleeding and other serious complications. Most of the current repair materials rely on imports. expensive

Method used

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  • Extraction method of bovine tendon type I collagen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1.1 Wash fresh beef Achilles tendon, remove fat, aponeurosis and muscle, cut into pieces (the size of the piece can be 0.5cm×0.5cm), add 10% sodium bicarbonate aqueous solution and soak for 16h (sodium bicarbonate The aqueous solution is not enough to cover the beef), take it out, wash it with deionized water, and dry it to obtain pure beef Achilles tendon;

[0026] 1.2 Add pure bovine Achilles tendon to a myristic acid solution containing 5% triethanolamine and soak for 2-3 hours, take it out, wash it with deionized water, and dry it;

[0027] The content of triethanolamine contained in each 1L of myristic acid solution is 0.15g;

[0028] 1.3 Add the above-mentioned dried beef Achilles tendon into liquid nitrogen, grind it into powder after it stops boiling, and obtain beef Achilles tendon powder;

[0029] 1.4 Take 20g of beef Achilles tendon powder, add 1000mL acetic acid-sodium acetate buffer solution, add pepsin accounting for 0.6% of the weight of the beef Achille...

Embodiment 2

[0033] 2.1 Wash fresh beef Achilles tendon, remove fat, aponeurosis and muscle, cut into pieces (the size of the piece can be 0.5cm×0.5cm), add 10% sodium bicarbonate aqueous solution and soak for 16h (sodium bicarbonate The aqueous solution is not enough to cover the beef), take it out, wash it with deionized water, and dry it to obtain pure beef Achilles tendon;

[0034] 2.2 Add the pure bovine Achilles tendon to a 6.5% myristic acid solution containing triethanolamine and soak for 2-3 hours, take it out, wash it with deionized water, and dry it in the air;

[0035] The content of triethanolamine contained in each 1L of myristic acid solution is 0.12g;

[0036] 2.3 Add the above-mentioned dried beef Achilles tendon into liquid nitrogen, grind it into powder after it stops boiling, and obtain beef Achilles tendon powder;

[0037] 2.4 Take 20g of beef Achilles tendon powder, add 1000mL acetic acid-sodium acetate buffer solution, add pepsin accounting for 0.8% of the weight of...

Embodiment 3

[0041] 3.1 Wash the fresh beef Achilles tendon, remove fat, aponeurosis and muscle, cut into pieces (the size of the piece can be 0.5cm×0.5cm), add 10% sodium bicarbonate aqueous solution and soak for 16h (sodium bicarbonate The aqueous solution is not enough to cover the beef), take it out, wash it with deionized water, and dry it to obtain pure beef Achilles tendon;

[0042] 3.2 Add the pure beef Achilles tendon to the myristic acid solution containing triethanolamine with a mass fraction of 8% and soak for 2-3 hours, take it out, wash it with deionized water, and dry it;

[0043] The content of triethanolamine contained in each 1L of myristic acid solution is 0.1g;

[0044] 3.3 Add the above-mentioned dried beef Achilles tendon into liquid nitrogen, grind it into powder after it stops boiling, and obtain beef Achilles tendon powder;

[0045] 3.4 Take 20g of beef Achilles tendon powder, add 1000mL acetic acid-sodium acetate buffer solution, add pepsin accounting for 0.7% of...

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Abstract

The invention belongs to the technical field of biomedical materials, and particularly relates to an extraction method of bovine tendon type I collagen. The extraction method comprises the following steps: washing fresh bovine tendons, removing fat, aponeuroses and muscles, cutting the bovine tendons into tendon blocks, soaking the tendon blocks with a sodium bicarbonate aqueous solution, cleaning, and airing to obtain pure bovine tendons; adding the pure bovine tendons in a triethanolamine-containing myristic acid solution for soaking, and cleaning and airing the soaked pure bovine tendons; adding the aired pure bovine tendons in liquid nitrogen, and grinding into powder to obtain bovine tendon powder; taking the bovine tendon powder, adding the bovine tendon powder in an acetic acid-sodium acetate buffer solution, adding pepsin and squalene, uniformly stirring, carrying out enzymatic hydrolysis, then adding glycol dimercaptoacetate, and carrying out oscillating reaction to obtain enzymolysis liquid; and carrying out salting-out and dialysis on the enzymolysis liquid to obtain the bovine tendon type I collagen. The preparation method provided by the invention is simple and the equipment requirement is low; and the obtained product is high-purity type I collagen with a triple helix structure, cannot cause mechanical injuries on nervous tissues, and is good in biocompatibility.

Description

technical field [0001] The invention belongs to the technical field of biomedical materials, and in particular relates to a method for extracting bovine Achilles tendon type I collagen. Background technique [0002] Collagen is the most abundant protein in vertebrates and one of the four major components of the extracellular matrix. It contains one or several regions of a triple helix structure composed of α chains. This unique triple helix structure and the covalent cross-linking between molecules endow collagen fibers with extremely high tensile strength and thermal stability. Structural basis for interaction with cells. [0003] In the existing extraction methods, the purity of the extracted collagen is low, and how to extract high-purity type I collagen with a triple helix structure has become a technical problem to be solved urgently. When the dura mater / dural mater is traumatized or invaded by tumors, it may cause different degrees of damage and defect of the meninge...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06
CPCC07K14/78C12P21/06
Inventor 司志华胡科
Owner QIANFOSHAN HOSPITAL OF SHANDONG
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