Method for preparing bioartificial heart valve material by cross-linking quercetin

A heart valve, quercetin technology, applied in the field of biomedical engineering, can solve problems such as fragility, brittleness, and weakening of the mechanical strength of biological valves

Inactive Publication Date: 2012-02-15
SHANGHAI INST OF CERAMIC CHEM & TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, glutaraldehyde itself has cytotoxicity, inhibits the completion of endothelialization of the valve host, affects the growth of normal human tissue, and results in a short working life of the valve; at the same time, the cytotoxicity and excessive cross-linking of glutaraldehyde itself also promotes valve calcification, Weaken the mechanical strength of the biological valve, making it brittle and fragile, greatly affecting the working period of the biological valve in the body [Schmidt CE and Jennie M. Baier. Acellular vascular tissues: natural biomaterials for tissue repair and tissue engineering. Biomaterials, 2000, 21 (22), 2215-223.]; In addition, although glutaraldehyde can kill bacteria, it cannot completely rule out the immune rejection of the recipient to the heterogeneous biological valve

Method used

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  • Method for preparing bioartificial heart valve material by cross-linking quercetin
  • Method for preparing bioartificial heart valve material by cross-linking quercetin
  • Method for preparing bioartificial heart valve material by cross-linking quercetin

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Effect test

Embodiment 1

[0056] Use the balanced salt buffer D-Hanks solution at 1-6°C as the storage and transportation solution, take the fresh porcine aortic heart valve from the slaughterhouse, cut the valve leaflets at 4°C within 4 hours, and use D-Hanks Fluid clean. Afterwards, the decellularized porcine heart aortic valve was prepared according to the aforementioned method, and washed cleanly to obtain the following: figure 1 Structure and shape of decellularized valve materials. Then add 2ml of quercetin to each valve to soak in ethanol-containing D-Hanks special solution, shake and cross-link at 120rpm at 37°C for 48 hours. Add 2ml of D-Hanks solution to each valve to soak the valve, shake and wash at 120rpm at 37°C for 48 hours, and repeat the washing 3 times. The obtained biological valve material was evaluated for the above performance. figure 2 Middle 2 is a decellularized valve cross-linked with 6.25 mg / ml glutaraldehyde, which is as loose as the non-cross-linked decellularized valve...

Embodiment 2

[0058] Prepare the decellularized porcine heart aortic valve according to the method described in the summary of the invention and clean it, add 2.5mg / ml quercetin-containing D-Hanks special solution with ethanol at a ratio of 2ml per valve. Example 1 Method Cross-linking, the resulting cross-linked biological valve material can also maintain the original shape of the valve, soft and moist without shrinking. image 3 The maximum tensile strength of the decellularized valve cross-linked by 2.5 mg / ml quercetin was 13.9 MPa. Figure 5 It was shown that only 28.5% of the decellularized valves cross-linked with 2.5 mg / ml quercetin were degraded after 24 hours of enzymatic hydrolysis. It shows that this concentration of quercetin also has a cross-linking effect on the decellularized valve, and has good mechanical properties and stability against enzymolysis.

Embodiment 3

[0060] Get fresh porcine aortic heart valve from slaughterhouse by the method of embodiment 1 and clean up. After washing with D-Hanks solution three times, valve cells were removed by shaking with decellularization solution at 60 rpm at 4°C for 96 hours. Use D-Hanks solution to shake the decellularized heart valve material at 21°C (room temperature) at 120rpm to wash valve cell residues, debris, free protein, nucleic acid and other macromolecules for 96 hours. Add 0.1mg / ml, 0.2mg / ml, 0.5mg / ml, 2mg / ml and 5mg / ml quercetin in ethanol-containing D-Hanks special solution at a rate of 20ml per valve, at 20°C, at 240rpm shaking speed, cross-linked for 96 hours. Add 20ml of D-Hanks solution to each valve to soak the valve, and wash at 120rpm for 10 minutes at room temperature. The bioprosthetic valve material of the present invention that makes like this is done aforementioned performance evaluation, and its maximum tensile strength is respectively 10.4MPa, 12.5MPa, 14.4MPa, 14.9M...

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Abstract

The invention discloses a cross-linking preparation method of an artificial biological heart valve material, which comprises a cross-linking agent, a cross-linking step and a preservation treatment after cross-linking. It is characterized in that after the collected porcine heart valve is processed and cleaned in time, the cells are removed, and after being fully cleaned, it is cross-linked with quercetin to improve its mechanical strength, further eliminate its antigenicity, and improve its stability. The pig decellularized heart valve material adopts the artificial biological heart valve prepared by the preparation method, which can make the valve non-toxic, high in tensile strength, resistant to enzymatic degradation, and release very little protein from the cross-linking agent and the valve itself. These cross-linking properties of the porcine decellularized heart valve material make it beneficial to the engraftment, proliferation, coverage and long-term anti-calcification of the human body's own valve interstitial cells and vascular endothelial cells after implantation.

Description

technical field [0001] The invention relates to a crosslinking preparation method of an artificial biological heart valve material, which belongs to the field of biomedical engineering. Background technique [0002] Worldwide, heart valve disease is associated with high morbidity and mortality. In the United States, 60,000 artificial heart valve replacement operations are performed every year, and about 20,000 people die directly from the disease [Julie R, BorisAN, Vacanti JP. Tissue engineering: a 21st century solution to surgical reconstruction. Ann thorac surg, 2001, 72 : 577-591.]. Clinically, artificial heart valves are mainly treated with mechanical valves and heterogeneous biological valves. Human allograft biological valves stored at low temperature are also used clinically, but they are rarely used due to insufficient sources. The coagulation effect of heterogeneous biological valves is weak, and patients do not need to take anticoagulant drugs after surgery. Co...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/38A61F2/24
Inventor 常江翟万银
Owner SHANGHAI INST OF CERAMIC CHEM & TECH CHINESE ACAD OF SCI
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