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Endothelialization biological heart valve and preparation method thereof

A heart valve and endothelialization technology, applied in the field of biomedical materials, can solve the problems of limited VEGF content and stability, and achieve the effects of improving endothelialization, prolonging service life, and reducing calcification

Active Publication Date: 2020-05-15
JILIN VENUS HAOYUE MEDICAL LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the content and stability of VEGF based on electrostatic loading is limited

Method used

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  • Endothelialization biological heart valve and preparation method thereof
  • Endothelialization biological heart valve and preparation method thereof
  • Endothelialization biological heart valve and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0022] A kind of endothelial biomaterial, it is made through the following steps:

[0023] (1) Obtain Human VEGFA transcript variant 4 ORF mammalian expression plasmamid, C-GFPSpark tag plasmid DNA and amplification: resuspend the plasmid lyophilized powder containing endothelial cell growth factor (VEGF), and thaw competent cells on ice; Add 2 μL of plasmid to 100 UL of melted competent cells, and then treat the plasmid-competent cell complex in ice bath for 30 min, heat shock at 42°C for 45 s, and ice bath for 2 min, and then add the complex to liquid culture without antibiotics Put it in the shaker at 37°C, and incubate at 200rmp for 60min; then take it out, centrifuge at 5000rmp for 5min, discard the supernatant, add 60μL of liquid medium containing antibiotics, resuspend and drop on the plate containing antibiotics overnight to cultivate. In a 20mL test tube, inoculate Escherichia coli carrying the desired plasmid into 5mL culture medium, and culture with shaking at 37°C...

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Abstract

The invention discloses an endothelialization biological material and a preparation method thereof. The preparation method comprises the following steps: amplifying a plasmid containing VEGF; extracting DNA of the VEGF plasmid; transfecting endothelial cells with the extracted plasmid DNA; mixing the transfected cells with a gel matrix, and coating the biological material with the mixture; and adding an endothelial cell suspension, culturing the material at the temperature of 37 DEG C for 20-26 h, and fixing the material with paraformaldehyde to obtain an endothelialization biological heart valve. According to the method, the heart valve material is prepared from hydrogel coating the VEGF plasmid DNA, so endothelialization of the biological material is improved, calcification is reduced, and the service life of the heart valve is prolonged.

Description

technical field [0001] The invention belongs to the technical field of biomedical materials, and in particular relates to an endothelialized biological material and a preparation method thereof. Background technique [0002] Bioprostheses currently used clinically are usually prepared from pig or bovine pericardium cross-linked with glutaraldehyde. However, glutaraldehyde-crosslinked biological valves still have the problem of calcification. One of the important reasons for the calcification of biological valves is the failure of endothelialization. On the body's normally functioning heart valves, endothelial cells prevent blood clotting and reduce calcification. Because glutaraldehyde has great cytotoxicity, glutaraldehyde-crosslinked biological valves are not conducive to the growth and proliferation of endothelial cells, resulting in their inability to endothelialize. [0003] Small interfering RNAs are used as sequence-specific gene silencing agents that primarily tar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/52A61L27/38A61L27/36C12N5/10
CPCA61L27/3687A61L27/3691A61L27/3808A61L27/52A61L2430/20C07K14/475C12N5/069C12N2510/00
Inventor 雷洋王云兵邓璐王刚
Owner JILIN VENUS HAOYUE MEDICAL LTD
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