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Method for fixing heparin multilayer film on the surface of titanium dioxide

A surface-fixed, multi-layer film technology, used in packaging food, packaging, medical containers, etc., can solve the problems of difficult to achieve anticoagulant effect, unstable silane coupling layer, poor binding force, etc., and achieve multi-reactive active sites. Spot, not easy to fall off, easy to dissociate effect

Inactive Publication Date: 2012-08-22
CO WITH LTD LIABILITY OF MEDICAL SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The prior art adopts a simple physical coating method to adsorb a layer of heparin on the surface of titanium-based metals. The binding force between the two is poor, and because heparin is water-soluble, it will be lost quickly in the blood flow environment, and it is difficult to achieve the expected resistance. coagulation effect
The chemical method of silane coupling on TiO 2 Heparin immobilized on the surface is also difficult to be applied clinically due to the instability of the silane coupling layer under physiological conditions

Method used

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  • Method for fixing heparin multilayer film on the surface of titanium dioxide
  • Method for fixing heparin multilayer film on the surface of titanium dioxide
  • Method for fixing heparin multilayer film on the surface of titanium dioxide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] A TiO 2 The method for fixing the heparin multilayer film on the surface, the steps are:

[0031] A. Preparation of biotinylated heparin

[0032] Add heparin, NHS (N-hydroxysuccinimide) and EDC (1-(3-dimethylaminopropyl) base)-3-ethylcarbodiimide), so that the ratio of the amount of carboxyl groups in heparin, NHS and EDC is 1:0.6:1, and then the ratio of the amount of carboxyl groups in hexamethylenediamine to heparin The ratio is 1.2:1, add hexamethylenediamine, and dialyze to remove small molecules after reaction to obtain heparin-hexamethylenediamine solution;

[0033] In addition, biotin with the same amount of substance as hexamethylenediamine was added to the MES solution with a concentration of 50mM, and then NHS and EDC were added to it in turn, so that the ratio of the three substances of biotin, NHS and EDC was 1: 0.6:1, the biotin activation solution is obtained;

[0034] Add biotin activation solution to heparin-hexamethylenediamine solution, dialyze to...

Embodiment 2

[0048] A TiO 2 The method for fixing the heparin multilayer film on the surface, the steps are:

[0049] A. Preparation of biotinylated heparin This step is exactly the same as step A in Example 1.

[0050] B. Self-assembly of organic phosphonic acid monolayers into clean anatase TiO 2 The material was soaked in the isooctane solution of monododecyl phosphate at a temperature of 120 °C and a concentration of 1 mmol / L for 1 hour, and the TiO 2 Monolayer self-assembly on the surface, taking out the TiO 2 Wash afterwards.

[0051] C. Immobilize biotin in a dark room, and the monolayer self-assembled TiO obtained in step B 2 , put the concentration of 0.3mg / ml azide biotin (photobiotin) in ethanol / water solution with a volume ratio of 3:0.5, after the surface is completely wetted, take it out, dry it in the shade, irradiate with ultraviolet light for 10min, and the radiation energy is 8mW / cm 2 , to immobilize biotin on TiO 2 surface.

[0052] D. Assemble the biotin-fixed ...

Embodiment 3

[0060] A TiO 2 The method for fixing the heparin multilayer film on the surface, the steps are:

[0061] A. Preparation of biotinylated heparin This step is exactly the same as step A in Example 1.

[0062] B. Self-assembly of organic phosphonic acid monolayers. Clean anatase TiO mixed with rutile 2 The material was immersed in the isooctane solution of monododecyl phosphate at a temperature of 50 °C and a concentration of 50 mmol / L for 18 hours, and the TiO 2 Monolayer self-assembly on the surface, taking out the TiO 2 Wash afterwards.

[0063] C. Immobilize biotin in a dark room, and the monolayer self-assembled TiO obtained in step B 2 , put the concentration of 0.1mg / ml azide biotin (photobiotin) in ethanol / water solution with a volume ratio of 3:0.1, after the surface is completely wetted, take it out, dry it in the shade, irradiate with ultraviolet light for 1min, and the radiation energy is 8mW / cm 2 , to immobilize biotin on TiO 2 surface.

[0064] D. Assemble ...

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Abstract

The invention discloses a method for fixing a heparin multilayer film on the surface of TiO2. By the adoption of a self-assembly monomolecular layer method, an interface layer which forms chemical bonding respectively with the surface of TiO2 and biotin is prepared between the surface of TiO2 and the biotin so as to improve the stability of fixing the heparin multilayer film. Hexamethylendiamine is used as a coupling agent and reacts respectively with the carboxyl of heparin and the carboxyl of the biotin to form biotinylated heparin, therefore a recognition site of avidin is available. The TiO2 modified by the self-assembly monomolecular layer is coated with biotin azide and the biotin is fixedly fixed on the surface of the TiO2 by a photochemistry method. The recognition reaction of biotin-avidin is utilized for assembling and forming the heparin multilayer film. The method can be used for fixedly fixing the heparin multilayer on a TiO2 thin layer at the surface of titanium-base biometal materials and on the rear surface of a TiO2 film deposited on the surface of other materials, thereby ensuring that the surface thereof can have excellent anticoagulant function. Furthermore, the method has simple technology and easy realization.

Description

Technical field [0001] The invention relates to a method for biochemical modification of the surface of inorganic materials, in particular to a method for modifying the surface of TiO 2 Method for Surface Immobilization of Heparin Multilayer Films. Background technique [0002] Titanium-based metals have been widely used in biomaterials, and a layer of oxide layer TiO will naturally form on the surface of titanium-based metal biomaterials. 2 , the TiO 2 layer proved to be the main reason for the good biocompatibility of titanium-based metal biomaterials, but the TiO on the surface of titanium-based metal biomaterials 2 The anticoagulant properties of thin layers are still insufficient. [0003] Heparin is an anticoagulant drug commonly used clinically. It achieves anticoagulant effect by terminating certain steps in blood coagulation. Immobilizing heparin on the surface of titanium-based metal materials can improve the anticoagulant performance of the materials. The pri...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L33/10
Inventor 黄楠翁亚军王进杨苹冷永祥陈俊英孙鸿万国江赵安莎游天雪吴熹
Owner CO WITH LTD LIABILITY OF MEDICAL SCI
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