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Method for crosslinking collagen through oxidized oligosaccharide

A technology for oxidizing oligosaccharides and collagen, applied in the field of biomedical materials and leather industry, can solve the problems of large molecular weight of oxidized polysaccharides, low cross-linking efficiency, large reaction steric hindrance, etc., and achieve good biocompatibility and high cross-linking efficiency. , the effect of small steric hindrance

Active Publication Date: 2016-01-20
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, oxidized polysaccharides have large molecular weight, large reaction steric hindrance, poor water solubility (such as oxidized chitosan, oxidized cellulose), poor permeability, and are difficult to penetrate materials with dense surfaces, which are prone to surface crosslinking and low crosslinking efficiency.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] (1) Weigh 5 g (dry weight) of acellular porcine dermal matrix into a conical flask, and add 80 ml of sodium carbonate-sodium bicarbonate buffer solution with pH 9.4;

[0033] (2) Weigh 1 g of oxidized seaweed oligosaccharide with an oxidation degree of 45%, add it to the above-mentioned Erlenmeyer flask, shake well, and close the mouth of the bottle;

[0034] (3) Place the above-mentioned Erlenmeyer flask in a water bath with a temperature-controllable cradle, adjust the reaction temperature to 37°C, and adjust the rotation speed to 150r / min;

[0035] (4) React for 24 hours, take out the Erlenmeyer flask, discard the reaction solution, add 50ml of 0.1mol / L ammonium chloride solution and shake for 0.5h, then discard the solution, add 100ml of water for injection to wash for 0.5 hours, discard and wash solution, repeated washing three times;

[0036] (5) Take out the cross-linked acellular porcine dermal matrix, and freeze-dry it in a freeze dryer.

Embodiment 2

[0038] (1) Weigh 3 grams of collagen film;

[0039] (2) Weigh 0.5 g of oxidized isomaltooligosaccharide with a degree of oxidation of 100%, place it in a beaker filled with 50 ml of phosphate buffer solution at pH 7.4, and shake it to completely dissolve it;

[0040] (3) Put the collagen film into the above beaker and close the mouth of the beaker;

[0041] (3) Put a magnetic stir bar into the above beaker, place the beaker on the magnetic stirrer, adjust the reaction temperature to 25°C, and the stirring speed to 1000r / min;

[0042] (4) After 40 hours, discard the reaction solution, add 50ml of distilled water to wash for 0.5 hours, and repeat the washing three times;

[0043] (5) Take out the cross-linked collagen membrane and air-dry it on an ultra-clean bench.

Embodiment 3

[0045] (1) Weigh 100 grams of collagen sponge, put it in a 30-liter reaction kettle, add 19 liters of 0.5mol / L acetic acid solution, and stir thoroughly at 4°C until the collagen sponge is completely dissolved;

[0046] (2) Weigh 10 grams of oxidized chitosan oligosaccharide with an oxidation degree of 10%, and dissolve it in 1 liter of 0.5mol / L acetic acid solution at a temperature of 4°C at 4°C;

[0047] (3) Slowly add the oxidized chitosan oligosaccharide solution into the collagen solution, and keep stirring at 4°C to carry out the cross-linking reaction;

[0048] (4) After reacting for 16 hours, use sodium hydroxide solution to adjust the pH value to be weakly alkaline (pH value is about 7.50); ​​slowly add solid ammonium sulfate to make the concentration about 1.5mol / L, stir continuously until it is completely dissolved, and then let stand Overnight (10-12h); centrifuge the next day (8000r / min, 15min), discard the supernatant, add 2 liters of 0.5mol / L acetic acid solutio...

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PUM

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Abstract

The invention relates to a method for crosslinking collagen through an oxidized oligosaccharide. The method comprises the steps that the oxidized oligosaccharide with a certain quantity of aldehyde groups serves as a crosslinking agent, a crosslinking reaction occurs between the oxidized oligosaccharide and collagen molecules or an aggregate of the collagen molecules, then cleaning or dialyzing treatment is performed, and finally freeze drying is performed with a freeze drying machine to obtain crosslinked collagen molecules containing the oligosaccharide structure or an aggregate of the crosslinked collagen molecules. According to the crosslinked collagen obtained through the method, not only are the original performances of collagen kept, but also the oligosaccharide structure with the activity is introduced, the formed crosslinked structure can resist a certain degradation function of biological enzymes, all the basic performances are excellent, the biocompatibility is good, and the certain functionality is simultaneously achieved. The method can be applied to biological material and leather industries.

Description

technical field [0001] The invention relates to a method for oxidizing oligosaccharides to cross-link collagen, which can be applied to biomedical materials and leather industries. Background technique [0002] Collagen is the main component of the extracellular matrix. It is the most abundant and widely distributed protein in animals. It is rich in animal skin, bones, tendons, visceral interstitial cells, ligaments, blood vessels, and sclera. Collagen has many advantages such as low immunogenicity, low irritation, low cytotoxicity, good biocompatibility, good performance of promoting cell growth and biodegradability, and is widely used in the field of biomedicine (but the health Hua, Wang Kunyu, Zeng Rui, et al. The medical application and development prospect of collagen [J]. Biomedical Engineering and Clinic, 2004, 8 (1): 45-52.). Collagen medical biomaterials for different purposes have different requirements on physical and mechanical properties, degradation rate, etc....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J3/24C08L89/00C08L5/00C08K5/1545
Inventor 但年华陈一宁但卫华
Owner SICHUAN UNIV
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