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Method of regulating degradation rate of porous collagen-based cradle with amino acid

A technology of degradation rate and porous scaffolds, applied in medical science, surgery, prosthesis, etc., can solve the problems of difficult regulation of cross-linking degree, cytotoxicity, and inability to meet the requirements of various tissues and organs with different regeneration rates, etc. Achieve the effects of simple preparation process, low cost, good biocompatibility and degradation performance

Inactive Publication Date: 2005-08-17
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of the traditional pure chemical cross-linking method is that it is usually difficult to control the degree of cross-linking and cannot meet the requirements of various tissues and organs with different regeneration rates; when glutaraldehyde is used as a cross-linking agent, the residual glutaraldehyde Can cause certain side effects, such as cytotoxicity

Method used

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  • Method of regulating degradation rate of porous collagen-based cradle with amino acid
  • Method of regulating degradation rate of porous collagen-based cradle with amino acid
  • Method of regulating degradation rate of porous collagen-based cradle with amino acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Effects of different kinds of amino acids on the degree of degradation of collagen-based porous scaffolds.

[0020] Type I collagen scaffolds (2.5 mg / block) derived from bovine tendon were placed in 24-well culture plates, and 1 ml of glycine (Gly), glutamic acid (Glu) or lysine (Lys) containing 2.5 μM 2-N-Morpholine ethanesulfonic acid (MES) solution (50 mM, pH 5.5), soaked for 1 hour; at the same time, three distilled water was added for soaking as a control. Then add 1ml of 1-ethyl-3-(dimethylaminopropyl)-carbodiimide (EDAC) containing 40mM, MES solution of 40mM N-hydroxysuccinimide (NHS) respectively, so that the final The concentrations of EDAC and NHS were 20 mM and 10 mM, respectively. Cross-link at room temperature for 24 hours, rinse with triple distilled water for 6 times, 10 minutes each time after cross-linking. After freeze-drying, the cross-linked collagen-based porous scaffold is obtained.

[0021] Add 3ml of 0.1mg / ml type I collagenase solution (PBS, ...

Embodiment 2

[0023] Effect of glutamic acid concentration on the degradation rate of collagen-based porous scaffolds.

[0024] Place the type I collagen scaffolds (2.5mg / block) derived from bovine tendon in a 24-well culture plate, add 1ml of 1-10μM glutamic acid MES solution (50mM, pH 5.5), soak for 1 hour; then Add 1 ml of MES solution containing 40 mM EDAC and 40 mM NHS respectively, so that the final concentrations of EDAC and NHS are 20 mM and 10 mM, respectively. Cross-link at room temperature for 24 hours, rinse with triple distilled water for 6 times, 10 minutes each time after cross-linking. After freeze-drying, the cross-linked collagen-based porous scaffold is obtained.

[0025] Take the cross-linked porous scaffolds added with different concentrations of glutamic acid, add 3ml of 0.5mg / ml type I collagenase solution (PBS, pH7.4), and digest in a constant temperature water bath at 37°C for 12h. Aspirate 1ml of the digested supernatant, put it in a polymerization tube, add 2ml ...

Embodiment 3

[0027] The effect of lysine concentration on the degradation rate of collagen-based porous scaffolds.

[0028] Place the type I collagen scaffolds (2.5mg / piece) derived from bovine tendon in a 24-well culture plate, add 1ml of 0.67-100μM lysine MES solution (50mM, pH 5.5), soak for 1 hour; then Add 1 ml of MES solution containing 40 mM EDAC and 40 mM NHS respectively, so that the final concentrations of EDAC and NHS are 20 mM and 10 mM, respectively. Cross-link at room temperature for 24 hours, rinse with triple distilled water for 6 times, 10 minutes each time after cross-linking. After freeze-drying, a cross-linked collagen-based porous scaffold is obtained.

[0029] Take the cross-linked porous scaffolds added with different concentrations of lysine, add 3ml of 0.5mg / ml type I collagenase solution (PBS, pH7.4), and digest in a constant temperature water tank at 37°C for 12h. Aspirate 1ml of the digested supernatant, put it in a polymerization tube, add 2ml of 6M hydrochlo...

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Abstract

Amino acid with high biocompatibility is used as cross-linking and bridging agent, amino acid in 0.1-100 microns is made to compound with porous collagen-based cradle, and the cradle is further cross-linking treated with carbonated diimine compound, so as to prepare porous collagen-based cradle with controllable degradation rate and good biocompatibility. The present invention uses material with wide source, low cost and no toxic side effect; and the preparation process is simple, feasible, mild in condition and good in repeatability. The prepared porous collagen-based cradle can meet the requirement for different tissue and organ to regenerate and may be used widely in regeneration and reconstruction of skin, cartilage, bone, blood vessel, nerve, tendon, heart valve and other organs in biomedicine, tissue engineering and other fields.

Description

technical field [0001] The invention relates to a method for using amino acid as a cross-linking bridging agent to regulate the degradation rate of a collagen-based porous scaffold under the action of the cross-linking agent. Background technique [0002] Tissue engineering involves many disciplines and fields such as medicine, chemistry, biology, and materials science. In the study of tissue engineering, the selection of scaffold materials and the construction of scaffolds are the key links. Collagen is the main component of connective tissue in mammals, constituting about 30% of the protein in the human body and 72% of the dry weight in the skin. There are 19 types of collagen, the most common being types I, II, and III. Among them, type I is the most abundant and has excellent performance, and is widely used in biomaterials. Although collagen materials have unparalleled biocompatibility, the mechanical strength of scaffolds constructed of pure c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/00A61L27/24A61L27/50A61L31/00
Inventor 高长有马列毛峥伟沈家骢
Owner ZHEJIANG UNIV
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