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Reagent combination and method for improving calcification resistance of bovine pericardial valve material

A bovine pericardium and valve technology, applied in the field of reagent combination to improve the anti-calcification performance of bovine pericardial valve materials, can solve problems such as insufficient glutaraldehyde cross-linking, and achieve the goal of improving anti-calcification performance, maintaining mechanical properties, and improving durability Effect

Active Publication Date: 2022-01-28
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the problem of insufficient glutaraldehyde cross-linking and only cross-linking of collagen, some solutions have been proposed at home and abroad, mainly by adding other cross-linking agents and glutaraldehyde to reduce calcification, such as joint Low-dose glutaraldehyde and other new cross-linking agents such as genipin, epoxy compounds, carbodiimides, etc., or adding new elastin and glycosaminoglycan cross-linking agents, the long-term effects of these methods are yet to be seen. Further observation and verification

Method used

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  • Reagent combination and method for improving calcification resistance of bovine pericardial valve material
  • Reagent combination and method for improving calcification resistance of bovine pericardial valve material
  • Reagent combination and method for improving calcification resistance of bovine pericardial valve material

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preparation example Construction

[0042] In the preparation method of the present invention, its key points are the combined use sequence of glutaraldehyde, neomycin, naringenin, magnesium chloride, at first remove cell component, remove most of antigen and phospholipid; Then apply neomycin (- NH 2 donor) to cross-link carboxyl groups (-COOH) to stabilize glycosaminoglycans; glutaraldehyde (-CHO) is then used to cross-link amino groups (-NH 2 ), ensure the mechanical properties, and block the antigen and immobilize neomycin; then use glutathione to block the residual aldehyde group (-CHO) to remove the residual toxicity of glutaraldehyde cross-linking; then add polyphenol cross-linking to stabilize elastin (Cross-linked -OH, -C=O, -NH 2 ), reduce immune and inflammatory responses; finally use metal ions to stabilize elastin and block the calcium ion binding sites of polyphenols, and at the same time change the surface charge properties of the material to competitively inhibit calcium ion deposition; through t...

Embodiment 1

[0051] Embodiment 1 Early stage treatment

[0052] Take out the pericardium from the cattle (about 400-500Kg in body weight) in the slaughterhouse, separate the fat and outer membrane within 30 minutes of warm ischemia time, and use sterile PBS solution containing antibiotics (penicillin and streptomycin, the concentration is 100U / ml) ) washed several times, immersed in 0.1% bromogeramine for 30 minutes, rinsed the inner and outer sides of the pericardium with sterile PBS containing antibiotics for 10 minutes×3 times, and stored at a low temperature of 4°C.

Embodiment 2

[0053] Example 2 Decellularization

[0054] Step 1: Treat 0.1% bromogeramine at 37°C for 30 minutes, stirring once every 10 minutes;

[0055] Step 2: Rinse with 0.25% TritonX-100 at 37°C and 120 rpm for 48 hours, changing the solution every 12 hours;

[0056] Step 3: 3u / ml DNase-I / 0.03mg / ml RNase-A, 37°C, 120rpm, shaking and drifting for 24h, changing the medium every 12h;

[0057] Step 4: Rinse with distilled water for 48 hours, and change the solution every 8 hours;

[0058] Step 5: Rinse with PBS for 24 hours, and store after changing the medium.

[0059] Through the above steps, the cellular components are removed, and most of the antigens and phospholipids are removed.

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Abstract

The invention relates to the field of biological materials, in particular to a reagent combination and a method for improving the calcification resistance of a bovine pericardial valve material. On the basis of the existing glutaraldehyde cross-linked decellularized bovine pericardium, decellularization and aldehyde group sealing are carried out, and meanwhile, elastin, a glycosaminoglycan cross-linking agent and a calcium ion inhibitor are supplemented for treatment. Compared with existing glutaraldehyde crosslinking, experiments prove that the calcification resistance can be improved on the basis of effectively maintaining the mechanical property, and the durability of the valve material is improved.

Description

technical field [0001] The invention relates to the field of biological materials, in particular to a reagent combination and a method for improving the anti-calcification performance of bovine pericardial valve materials. Background technique [0002] At present, biological valves prepared by cross-linking heterogeneous (pig or bovine) pericardium are the mainstream products in the market. The valve leaflets of the interventional biological valves produced by Edward Life Sciences of the United States, Medtronic of the United States, and Hangzhou Qiming Medical Devices Co., Ltd. are all made of pig or bovine pericardium raw materials and cross-linked with glutaraldehyde. However, the current lifespan of bioprosthetic valves cross-linked with heterogeneous pericardium glutaraldehyde is usually only about 10-15 years, which greatly limits the application range and effect of bioprosthetic valves. [0003] The failure of biological valves is caused by many factors, including ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N23/2251G01N23/2202G01N1/31G01N1/36
CPCG01N23/2251G01N23/2202G01N1/31G01N1/36G01N2001/366
Inventor 姜振林宋明哲吴忠仕齐晓科唐贞洁刘育宏
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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