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Construction method for skin tissue engineering rack containing epidermal growth factor

A technology of epidermal growth factor and skin tissue engineering, applied in medical science, dressings, prostheses, etc., can solve the problems of poor elasticity of collagen sponge membrane, reduce collagen coagulation ability, poor stability, etc., and achieve good flexibility and enzyme resistance , the effect of inducing autologous cell growth and inducing differentiation and growth

Inactive Publication Date: 2004-07-14
CHINA LEATHER & FOOTWEAR IND RES INST +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The collagen sponge membrane formed by freeze-drying the collagen solution has poor elasticity, is easily broken, and has poor stability, so it needs to be cross-linked by adding a cross-linking agent or undergoing special treatment.
[0006] Among the existing technologies that use collagen as raw material to make collagen sponge membranes as wound dressings, some use glutaraldehyde as a crosslinking agent to improve the strength, toughness and stability of collagen sponge membranes, but reports have confirmed that Dialdehyde has obvious cytotoxicity. When cells are cultured with a sponge membrane containing glutaraldehyde, there is little cell ingrowth, and as the collagen sponge membrane gradually degrades in the body, glutaraldehyde monomers will also be released into body fluids Among them, it is harmful to the human body, so glutaraldehyde is not an ideal cross-linking agent; some use chondroitin sulfate as a cross-linking agent to improve the strength of the collagen sponge membrane and the tolerance to enzymes, but chondroitin sulfate and collagen Molecules can only be bonded with weak bonds, which has the disadvantages of promoting scar formation and reducing collagen coagulation ability, and is also not an ideal cross-linking agent

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The fetal cowhide is depilated, fascia, fat and blood stains are removed according to the conventional method, washed, frozen, cut into thin slices with a thickness of 0.5mm, dehydrated with acetone three times, degreased with petroleum ether three times, and then vacuumed to remove petroleum ether to make dehydrated Degreased fetal cowhide is used as a raw material, 10 g of the raw material is taken, washed with double distilled water and put into a reactor, and an aqueous solution prepared from 0.1 g of gastric enzyme and 100 g of water is added. Carry out an enzyme treatment, at 20°C and pH=2, mechanically shake for 24 hours to degrade the covalent bonds between tissue fibers, centrifuge, discard the supernatant, take the precipitate, wash three times with double distilled water, add 500g of 0.5M acetic acid aqueous solution, at 0°C, mechanically vibrate for 32 hours, then add sodium chloride to make the concentration of sodium chloride in the solution reach 2M, after...

Embodiment 2

[0026]Carry out dehydration and degreasing of the bovine Achilles tendon according to the method of Example 1, vacuumize and remove petroleum ether, and make the dehydrated and degreasing bovine Achilles tendon as raw material, take 10 g of the raw material and wash it with double distilled water and add it to the reactor, add 0.3 g of pancreatic Perform an enzyme treatment with chymotrypsin and 100g of the prepared aqueous solution. Under the conditions of 30°C and pH=6, mechanically shake for 16 hours to degrade the covalent bonds between tissue fibers. After centrifugation, discard the supernatant, take the precipitate, and use double Wash with distilled water four times, add 1000g 0.5M acetic acid aqueous solution, at 10°C, mechanically vibrate for 28 hours, then add chlorine and sodium to make the concentration of sodium chloride in the solution reach 2M, after the precipitation is complete, centrifuge, discard the supernatant, Take the precipitate, put it into a dialysis ...

Embodiment 3

[0028] Use the bovine Achilles tendon dehydrated and degreased in Example 2 as raw material, get 10g, add in the reactor after washing with double distilled water, add the aqueous solution that is prepared by 0.5g pineapple enzyme and 100g water and carry out enzymatic treatment once, at 40 ℃, pH Under the condition of =8, mechanically vibrate for 8 hours to degrade the covalent bonds between tissue fibers, centrifuge, discard the supernatant, take the precipitate and wash it four times with double distilled water, add 1500g of 0.5M acetic acid aqueous solution, and heat at 20°C After mechanical vibration for 24 hours, add sodium chloride to make the concentration of sodium chloride in the solution reach 2M. After the precipitation is complete, centrifuge, discard the supernatant, take the precipitate and put it into a dialysis bag for dialysis five times, and collect the dialysis collagen Add the solution into the reactor, add 0.1g chymotrypsin and 100g water for secondary enz...

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Abstract

The construction process of skin tissue engineering rack containing epidermal growth factor with fetus calf or ox tendo achillis as basic material includes dewatering, defatting, primary enzyme treatment to reduce the covalent bond among tissue fibers, dialysis, secondary enzyme treatment to remove antigenic determinant, dissolving with acetic acid to obtain no-antigenicity collagen solution and collagen sponge film, soaking in propanetriol to adsorb, drying, mixing water solution of heparin and acetic acid, water solution of epidermal growth factor and propanetriol wetted collagen sponge film, and final freeze drying. The rack has no antigenicity, high strength, high flexibility, convenient operation implantation, no toxicity to wound, and capacity of inducing wound cell growth and promoting wound healing.

Description

technical field [0001] The invention belongs to the class of dressings for medical devices, specifically a method for constructing a skin tissue engineering scaffold by adopting a tissue engineering method and taking fetal cowhide and bovine Achilles tendon collagen as scaffolds. Background technique [0002] With the emergence and development of tissue engineering science, research on skin wound repair dressings has shifted from pure wound dressing research to active artificial skin research. Active artificial skin constructed by tissue engineering has become the best choice for skin wound repair. [0003] The research on skin tissue engineering scaffold is one of the focuses of tissue engineering research. It has a history of nearly 20 years. In-depth studies have shown that although polyesters have good biological properties, the degradation products will increase the acidity of the surrounding tissues, which is limited in use; natural biomaterials mainly composed of coll...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61F2/02A61F13/02A61L27/60
Inventor 杨颖颙于淑贤刘杰王德文
Owner CHINA LEATHER & FOOTWEAR IND RES INST
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