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Straight-chain heparin modified biotype artificial blood vessel

A bio-based, artificial blood vessel technology, applied in the types of packaging items, anti-coagulation treatment, special packaging items, etc., can solve the problem that the regeneration and remodeling process is difficult to occur, and the surface properties cannot meet the needs of biomaterial compatibility and tissue regeneration. , Heparin activity is difficult to guarantee, etc.

Active Publication Date: 2013-05-08
PEKING UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, despite the natural structure and composition of acellular matrices, their surface properties may still not meet the needs of biomaterial compatibility and tissue regeneration.
The small-diameter blood vessels based on acellular matrix (that is, biological blood vessels) currently have the following problems: thrombus generally forms within half a year; endothelialization of the inner surface of blood vessels and regeneration and remodeling of implantable materials by vascular smooth muscle cells are difficult to occur
[0013] 2. The inner surface of blood vessels can be modified by using the functional groups of natural materials, but the current method generally needs to directly react with functional groups such as heparin carboxyl groups and material surface amino groups. This reaction method is difficult to have a clear heparin modification site, and the activity of heparin is difficult. ensure

Method used

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  • Straight-chain heparin modified biotype artificial blood vessel
  • Straight-chain heparin modified biotype artificial blood vessel
  • Straight-chain heparin modified biotype artificial blood vessel

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1, Preparation of terminal cystamined heparin and terminal thiolated heparin

[0058] 1. Preparation of 3-5K and 5-10K heparin by enzymatic degradation

[0059] Heparin extracted from porcine small intestinal mucosa is degraded by type I heparanase for a certain period of time, and then ultrafiltered and dialyzed with an ultrafiltration membrane with strong molecular weight selectivity to obtain:

[0060] (1) Prepare 0.1M Tris-hydrochloric acid buffer solution, and adjust the pH between 7.00±0.02.

[0061] (2) Weigh 806.9 mg of heparin powder (white), and sterilize it by ultraviolet light for 30 minutes in a biological safety cabinet. This was added to 16.0 ml of Tris buffer filtered through a 220 micron pore filter. (take a sample and measure the pH value of the system with a pH meter, it is 7.08).

[0062] (3) Filter the bacteria with a filter membrane again. Add 100 μl of 100 units (sigma unit, about 1 / 600th international unit) heparinase-Tris buffer solu...

Embodiment 2

[0103] Example 2: Heparin compounded on the acellular matrix artificial blood vessel

[0104] After reducing the end of cystamined heparin, thiolated heparin is obtained, and then quantitatively detects the amount of amino groups on the inner surface of blood vessels, and then activates the inner surface of blood vessels through a linker with a thiol-amino bidirectional functional group, and then covalently immobilizes the heparin with a direct chemical bond. Chain-shaped heparin on the inner surface of the vascular material (see Figure 4 ).

[0105] 1. Detection of amino groups on blood vessels

[0106] Reaction principle: Molecules containing primary amine or hydrazide groups can react with

[0107] 2,4,6-trinitrobenzenesulfonate (TNBS) reacts to generate colored derivatives; the amino group content in the complex is linearly related to the absorption value of the orange-yellow product at 335nm. Therefore, the amino group content can be determined by measuring the absorb...

Embodiment 3

[0150] Example 3. Characterization of Anticoagulation Properties of Composite Artificial Blood Vessels

[0151] A few milliliters of blood is collected from the vein of healthy New Zealand rabbits, and 10% of the total volume of sodium citrate is added, which can have an anticoagulant effect in a short time. The collected fresh rabbit whole blood was centrifuged at 1500rpm in a common centrifuge for 15min, then the upper and middle liquids were drawn into another centrifuge tube, centrifuged again at 3000rpm for 10min, and after centrifugation, 75% of the supernatant was drawn and preserved, which was platelet-poor plasma (PPP ), and the remaining fluid is platelet-rich plasma (PRP). Among them, PPP is used for the detection of plasma recalcification time, prothrombin time, and activated partial thromboplastin time, and PRP is used for platelet adhesion test.

[0152] 1. Platelet adhesion assay (PAA)

[0153] 1) Each blood vessel sample (1cm 2 , refers to the double-sided s...

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Abstract

The invention discloses straight-chain terminal cystamine heparin and a biotype artificial blood vessel modified by the same. The structural formula of the terminal cystamine heparin provided by the invention is shown as a formula I. A preparation method of the straight-chain heparin modified biotype artificial blood vessel comprises the following steps of: cutting the disulfide bond in the terminal cystamine heparin by a reducing agent to obtain a sulfydryl locus capable of reacting with a blood vessel stent material; activating the inner surface of the blood vessel by a coupling agent with a bidirectional functional group; and fixing the heparin in a straight-chain shape on the inner surface of the blood vessel material through a chemical bond in a covalent manner so as to obtain a compound type artificial blood vessel with an anticoagulation effect. Compared with the existing method for modifying material surface through heparin carboxyl, the terminal locus reaction maintains the natural characters of heparin molecules to the greatest extent. When the straight-chain heparin is modified on the inner surface of the blood vessel, the resistance of the inner wall of the blood vessel against the platelet adhesion is improved, the anticoagulation performance of the blood vessel is obviously enhanced, and thus the biotype artificial blood vessel is superior to the blood vessel before modification.

Description

technical field [0001] The invention relates to a biological artificial blood vessel modified by linear heparin. Background technique [0002] Artificial blood vessels are mainly used for replacement repair of human tissue blood vessels. At present, large-caliber artificial blood vessels based on artificial materials (such as polyester and polytetrafluoroethylene) have been used clinically, while small-caliber artificial blood vessels (diameter less than 6mm) have not been clinically produced. The main reasons are the slow blood flow and low blood pressure of small-caliber vessels, which are prone to acute thrombosis, anastomotic intimal hyperplasia, aneurysm, infection and atherosclerosis. In current clinical treatment, the substitutes for small-caliber vascular grafts are mostly autologous blood vessels (such as vascular tissue from the great saphenous vein). However, many patients may not be able to provide transplantable blood vessels due to other vascular diseases or l...

Claims

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Application Information

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IPC IPC(8): C08B37/10A61L33/10
Inventor 罗莹刘晓鹏张伟
Owner PEKING UNIV
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