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Construction method of artificial blood vessel

A construction method and artificial blood vessel technology are applied in the field of construction of polyester artificial blood vessels, which can solve the problems such as the inability to use small-diameter artificial blood vessels for preparation, and achieve the effects of protecting blood cells, having a single molecular weight and preventing thrombus blockage.

Active Publication Date: 2016-02-17
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Aiming at some problems in the clinical application of polyester artificial blood vessels and the fundamental problem that it cannot be applied to the preparation of small-diameter artificial blood vessels, the present invention aims to provide a polyester surface-introduced with hydrophilic and negatively charged polypeptides and cell adhesion-promoting polypeptides. A method for constructing artificial blood vessels. The constructed artificial blood vessels are hemocompatible materials with excellent surface hydrophilicity, negative charge and endothelialization potential, which are beneficial to tissue healing and anticoagulation

Method used

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Experimental program
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Effect test

Embodiment 1

[0022] 1. Transfect Escherichia coli (BL21) cells with the prokaryotic system expression vectors carrying the first functional polypeptide gene and the second functional polypeptide gene respectively, coat them on Luria-Bertani solid medium containing ampicillin, and place them upside down at 37°C cultured in a biochemical incubator for 14-16 hours; pick a single colony and put it into 4 mL of Luria-Bertani liquid medium containing fresh ampicillin, and insert it into an air shaker with a shaking speed of 200r / min at 37°C for 8-10 hours; The cultured bacterial liquid was amplified and cultured in 1L of fresh Luria-Bertani liquid medium containing ampicillin at a ratio of 1:100; when the bacterial liquid density reached OD 600 When = 0.3-1.8, 0-1.2 mM isopropyl-β-D-thiogalactoside was added to induce culture for 0-8 hours, and the bacterial cells were collected by centrifugation at 4°C.

[0023] 2. Resuspend and mix the collected bacterial cells with the binding buffer of gluta...

Embodiment 2

[0031] 1. Transfect Escherichia coli (BL21) cells with the prokaryotic system expression vectors carrying the first functional polypeptide gene and the second functional polypeptide gene respectively, coat them on Luria-Bertani solid medium containing ampicillin, and place them upside down at 37°C Incubate in a biochemical incubator for 14-16 hours; pick a single colony and put it into 4 mL of Luria-Bertani liquid medium containing fresh ampicillin, and insert it into an air shaker with a shaking speed of 200r / min at 37°C for 8-10 hours; The cultured bacterial liquid was amplified and cultured in 1L of fresh Luria-Bertani liquid medium containing ampicillin at a ratio of 1:100; when the bacterial liquid density reached OD 600 When = 0.3-1.8, 0-1.2 mM isopropyl-β-D-thiogalactoside was added to induce culture for 0-8 hours, and the bacterial cells were collected by centrifugation at 4°C.

[0032] 2. Resuspend and mix the collected bacterial cells with the binding buffer of gluta...

Embodiment 3

[0040] 1. Transfect Escherichia coli (BL21) cells with the prokaryotic system expression vectors carrying the first functional polypeptide gene and the second functional polypeptide gene respectively, coat them on Luria-Bertani solid medium containing ampicillin, and place them upside down at 37°C Incubate in a biochemical incubator for 14-16 hours; pick a single colony and put it into 4 mL of Luria-Bertani liquid medium containing fresh ampicillin, and insert it into an air shaker with a shaking speed of 200r / min at 37°C for 8-10 hours; The cultured bacterial liquid was amplified and cultured in 1L of fresh Luria-Bertani liquid medium containing ampicillin at a ratio of 1:100; when the bacterial liquid density reached OD 600 = 0.3-1.8 0-1.2 mM isopropyl-β-D-thiogalactoside was added to induce culture for 0-8 hours, and the bacterial cells were collected by centrifugation at 4°C.

[0041] 2. Resuspend and mix the collected bacterial cells with the binding buffer of glutathione...

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Abstract

The invention discloses a construction method of an artificial blood vessel, and the method particularly includes the step that both the interior and the exterior of a polyester knitting tube are evenly coated with regenerated silk fibroin introducing two types of functional peptides. According to one of the functional peptides, the chain side base contains a great number of hydrophilic carboxy groups, and the acidic amino acid and molecules of the peptide are of electronegativity; the other peptide contains eight RGD and promotes cell adhesion. The two types of peptides come from expressions of living bodies, the sequences are analogues from natural protein, no toxicity or irritation exists, the molecular weight is singular, the peptides and the silk fibroin can be arranged on a polyester pipe in a covalent binding and coated mode, the hydrophilic performance and electronegativity are provided for the artificial blood vessel continuously and stably, endothelialization is promoted, harm to cells is lowered, and thrombus blockage caused by proteinosis and blood cell aggregation is prevented. The constructed artificial blood vessel has good biocompatibility and has an electronegative membrane layer similar to a natural blood vessel and a microenvironment for promoting adhesion growth of endothelial cells, and accordingly it is beneficial to protect blood cells and prevent thrombus from forming.

Description

technical field [0001] The invention relates to the field of polyester artificial blood vessels used in the replacement of vascular lesions, in particular to a method for constructing polyester artificial blood vessels with hydrophilic and negatively charged polypeptides and cell adhesion-promoting polypeptides introduced into the surface. Background technique [0002] There are millions of deaths due to cardiovascular and cerebrovascular diseases in my country every year, and the proportion is increasing year by year by about 30%. Therefore, vascular transplantation has become a hot spot of concern. Artificial blood vessels are currently the most scarce medical devices in clinical practice. Among them, the transplantation of small-caliber artificial blood vessels is still a clinical blank. Even for medium and large-caliber artificial blood vessels, there are very few products in my country. The annual use ratio of domestic products is very small, only 20 %about. It is conse...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/34A61L27/50
CPCA61L27/34A61L27/507A61L2420/06
Inventor 王建南刘云飞郝云霞裔洪根
Owner SUZHOU UNIV
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