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

A degradation rate, porous scaffold technology, used in medical science, prosthesis, surgery, etc., can solve problems such as difficulty in regulating cross-linking degree, cytotoxicity, and inability to meet the requirements of various tissues and organs with different regeneration rates. To achieve the effect of simple and easy preparation process, low cost, good biocompatibility and degradation performance

Inactive Publication Date: 2003-11-05

ZHEJIANG UNIV

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AI-Extracted 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 t...

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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 withwide 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 inbiomedicine, tissue engineering and other fields.

Application Domain

SurgeryProsthesis

Technology Topic

ChemistryTissue engineering +10

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Examples

Experimental program(6)

Example Embodiment

[0018] Example 1

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

[0020] Place the type I collagen scaffold (2.5mg/piece) derived from beef tendon in a 24-well culture plate, and add 1ml of 2.5μM glycine (Gly), glutamic acid (Glu) or lysine (Lys) respectively. 2-N-morpholine ethane sulfonic acid (MES) solution (50 mM, pH 5.5), soak for 1 hour; at the same time, add three-distilled water as a control. Then add 1ml of MES solution containing 40mM 1-ethyl-3-(dimethylaminopropyl)-carbodiimide (EDAC) and 40mM N-hydroxysuccinimide (NHS) to make the final The concentrations of EDAC and NHS are 20mM and 10mM, respectively. Cross-link at room temperature for 24 hours, after cross-linking, rinse with three-distilled water 6 times for 10 minutes each time. After freeze-drying, the cross-linked collagen-based porous scaffold is obtained.

[0021] Take a collagen-based porous scaffold without cross-linking, no amino acid cross-linking, and different amino acid cross-linking, add 3ml of 0.1mg/ml type I collagenase solution (PBS, pH7.4), and digest in 37℃ constant temperature water tank. 12h. 1ml of the supernatant was removed by aspiration, placed in a polymerization tube, 2ml of 6M hydrochloric acid was added, and the tube was sealed and hydrolyzed at 120°C for 12 hours. The degradation degree of the collagen porous scaffold was characterized by measuring the content of hydroxyproline in the hydrolysate. The degradation degree of porous scaffolds without cross-linking, without adding amino acid cross-linking, and adding different amino acid cross-linking can be seen figure 1.

Example Embodiment

[0022] Example 2

[0023] The effect of the concentration of glutamic acid on the degradation rate of the collagen-based porous scaffold.

[0024] Place the type I collagen scaffold (2.5mg/piece) 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 1 ml of MES solution containing 40 mM EDAC and 40 mM NHS was added to make the final EDAC and NHS concentrations respectively 20 mM and 10 mM. Cross-link at room temperature for 24 hours, after cross-linking, rinse with three-distilled water 6 times for 10 minutes each time. After freeze-drying, the cross-linked collagen-based porous scaffold is obtained.

[0025] Take the porous scaffolds cross-linked with different concentrations of glutamic acid, add 3 ml of 0.5 mg/ml type I collagenase solution (PBS, pH 7.4), and digest in a constant temperature water tank at 37° C. for 12 hours. 1ml of the supernatant was removed by aspiration, placed in a polymerization tube, 2ml of 6M hydrochloric acid was added, and the tube was sealed and hydrolyzed at 120°C for 12 hours. The degradation degree of the collagen porous scaffold was characterized by measuring the content of hydroxyproline in the hydrolysate. The effect of the concentration of glutamic acid on the degradation of collagen-based porous scaffolds figure 2.

Example Embodiment

[0026] Example 3

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

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

[0029] Take the porous scaffold cross-linked with different concentrations of lysine, add 3ml of 0.5mg/ml type I collagenase solution (PBS, pH7.4), and digest it in a constant temperature water tank at 37°C for 12h. 1ml of the supernatant was removed by aspiration, placed in a polymerization tube, 2ml of 6M hydrochloric acid was added, and the tube was sealed and hydrolyzed at 120°C for 12 hours. The degradation degree of the collagen porous scaffold was characterized by measuring the content of hydroxyproline in the hydrolysate. The effect of the concentration of lysine on the degradation of collagen-based porous scaffolds is seen image 3.

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Classification and recommendation of technical efficacy words

No side effects
Good biocompatibility and degradability

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

AI-Extracted Technical Summary

Problems solved by technology

Abstract

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Examples

Example Embodiment

Example Embodiment

Example Embodiment

PUM

Description & Claims & Application Information

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