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Method for preparing medical level type II collagen

A collagen and raw material technology, applied in the field of preparation of medical grade II collagen, can solve the problems of type II collagen precipitation, destruction of amino acid components, serious equipment corrosion, etc., and achieve no denatured collagen impurities, low antigenicity, and sterile finished products Effect

Inactive Publication Date: 2008-11-12
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the acid hydrolysis and alkali hydrolysis methods in the crude extraction method are rapid and thorough, they will destroy the amino acid components in the collagen, and the yield is low, the equipment is severely corroded, and secondary pollution occurs; the DEAE fiber column ion exchange chromatography method in the purification method consumes long time, low purification efficiency, poor repeatability, and precipitation of the purified type II collagen; although high-performance liquid ion-exchange chromatography and polyacrylamide gel electrophoresis guarantee the quality of the purified collagen, they can only be used for Micro-purification, not suitable for mass production

Method used

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  • Method for preparing medical level type II collagen
  • Method for preparing medical level type II collagen
  • Method for preparing medical level type II collagen

Examples

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Effect test

Embodiment 1

[0034] The preparation method of embodiment 1 medical grade type II collagen

[0035] 1. Pretreatment of articular cartilage: Add 2000ml of mixed solution of ethanol: chloroform=0.5 volume ratio to 200g of fresh articular cartilage to soak and degrease, replace the above solution every 5 hours, pour out the solution in 15 hours, and use a concentration of 99% Soak in 2000ml of ethanol aqueous solution, replace once every 15 minutes, pour off the 99% ethanol aqueous solution in 1 hour, wash once with 2000ml ethanol aqueous solution with a concentration of 50%, then soak with 2000ml NaCl aqueous solution with a concentration of 10%, replace once every 15 minutes , pour off the NaCl aqueous solution with a concentration of 10% in 1 hour, wash it twice with 2000ml ultrapure water, then soak the cartilage in 2000ml ultrapure water overnight, pour off the ultrapure water, put it into a beater to beat the articular cartilage Slurry;

[0036] 2. Extraction of proteoglycans with guani...

Embodiment 2

[0044] Example 2 Preparation method of medical grade type II collagen.

[0045] 1. Pre-treatment of articular cartilage: add ethanol: chloroform=1.5 volume ratio mixed solution 5000ml in fresh articular cartilage 300g, soak and degrease, change above-mentioned solution once every 15 hours, pour out solution in 45 hours, use concentration to be 99% Soak in 5000ml of ethanol aqueous solution, replace once every 30 minutes, pour off the 99% ethanol aqueous solution in 2 hours, wash once with 5000ml ethanol aqueous solution with a concentration of 80%, then soak with 5000ml NaCl aqueous solution with a concentration of 30%, replace once every 30 minutes , pour off the NaCl aqueous solution with a concentration of 30% in 2 hours, wash it twice with 5000ml of ultrapure water, then soak the cartilage in 5000ml of ultrapure water overnight, pour off the ultrapure water, and put it into a beater to beat the articular cartilage Slurry;

[0046] 2. Extraction of proteoglycan with guanid...

Embodiment 3

[0054] Example 3 Preparation method of medical grade type II collagen.

[0055] 1. Pre-treatment of articular cartilage: Add 3000ml of mixed solution of ethanol:chloroform=1 volume ratio to 250g of fresh articular cartilage to soak and degrease, replace the above solution every 10 hours, pour off the solution in 30 hours, and use a concentration of 99% Soak in 3000ml of ethanol aqueous solution, replace once every 20 minutes, pour off the 99% ethanol aqueous solution in 80 minutes, wash once with 3000ml ethanol aqueous solution with a concentration of 70%, then soak with 3000ml NaCl aqueous solution with a concentration of 20%, replace it for 20 minutes Once, pour out the NaCl aqueous solution with a concentration of 20% in 80 minutes, wash it twice with 3000ml of ultrapure water, then soak the cartilage in 3000ml of ultrapure water overnight, pour off the ultrapure water, and place the articular cartilage in a beater pulped;

[0056] 2. Extraction of proteoglycans with guani...

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Abstract

The invention discloses a method for making medical II collagen, which is characterized in: doing pre-treatment by using fresh articular cartilage as raw material; then using hydrochloric acid carbamidine to extract proteoglycan which comprises preparing a 4M hydrochloric acid carbamidine solution, and regulating the pH value to between 6.5 and 7.5 with alkali, then pouring the Hydrochloric acid carbamidine solution which is 8 to 15 times of the articular cartilage plasma into the cartilage plasma and blend, placing the solution on the shaking bed, shaking for one or two days, centrifuging and obtaining a solid matter, removing residue hydrochloric acid carbamidine solution by ultra-pure water washing for several times; pepsin digestion, which comprises that 2000 to 5000 portions of 0.5 to 0.6M acetic acid solution is added in the solid matter, then 0.1 to 0.3 portions of 2000 to 5000 mass percent solid matter which is pepsin is added in twice , for the first time half of the total amount is added and shaken in the shaking bed for 24 hours ,for the second time the other half is added and shaken for 24 hours, centrifuged, and the supernatant is removed, added in 0.01 to 0.02M aqueous solution of EDTA and 1 to 4 inactivate pepsin; salted out with NaCl; dialyzed with Na2 HPO4 solution, then medical-grade II Collagen is obtained.

Description

technical field [0001] The invention relates to a preparation method of medical grade type II collagen, which belongs to the field of preparation of biomedical materials. Background technique [0002] Type II collagen (Type II Collagen, C II) is the main organic component in cartilage matrix and vitreous fluid. It can strengthen the skin's water retention capacity, fill the gaps between collagen fibers in the skin, and maintain the morphology and tensile strength of articular cartilage. Its unique structure and chemical composition make it implantable in the body, drug controlled release carrier, tissue Engineering scaffolds, pathological research, pharmaceuticals, cosmetics and food are widely used. Recent studies have shown that type II collagen plays an important role in the pathological research of rheumatoid arthritis and bone tumors (Snowden N, Reynolds I, Morgan K, et al. T cell responses to human type II collagen in patients with rheumatoid Arthritis and health con...

Claims

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Application Information

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IPC IPC(8): A61L27/24A61L31/04A61K47/42A61K38/39A61K8/65A23L1/29A23L1/30A23L33/17
Inventor 蒋波张焰王文杰朱楚洪樊渝江
Owner SICHUAN UNIV
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