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Method for constructing Sema 4D/Heparin microenvironment on surface of cardiovascular implanted material

An implant material, cardiovascular technology, applied in pharmaceutical formulations, coatings, pharmaceutical sciences, etc., can solve problems such as no discovery, and achieve the effect of delaying half-life

Active Publication Date: 2017-03-08
SOUTHWEST JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Heparin (Heparin) is often used as an anticoagulant clinically, mainly for the treatment of thrombosis, thrombophlebitis and thromboembolism; the structure of heparin contains a large number of sulfate groups, and it is the most negatively charged biological substance found so far. Molecule; Sema4D is an anchor protein on the surface of cell membranes, widely present on the surface of platelets, T cells, etc., and its extracellular protein hydrolyzate Sema 4D can promote endothelial cell migration and immune regulation; Sema 4D receptors include plexin B1 and CD72, The former mainly exists on the surface of endothelial cells, while the latter mainly exists in immune cells. The function of Sema 4D is mainly realized by acting on the corresponding receptors; the PI of Sema 4D is 8.5, and it presents a positive charge under neutral pH conditions. Combination of effects; studies have shown that the combination of heparin and various growth factors can protect growth factors, delay the half-life of growth factors, and promote the recognition and combination of growth factors and their receptors; so far, no combination of heparin and Sema 4D combines technologies for cardiovascular materials

Method used

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  • Method for constructing Sema 4D/Heparin microenvironment on surface of cardiovascular implanted material
  • Method for constructing Sema 4D/Heparin microenvironment on surface of cardiovascular implanted material
  • Method for constructing Sema 4D/Heparin microenvironment on surface of cardiovascular implanted material

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Experimental program
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Effect test

Embodiment 1

[0044] A method for constructing a Sema 4D / Heparin microenvironment on the surface of a cardiovascular implant material, comprising the following steps:

[0045] A. Alkali activation

[0046] After the pure titanium is polished and cleaned, it is activated with 1mol / L NaOH for 12 hours, after ultrasonic cleaning with single distilled water, it is immersed in double distilled water, and reacted at 80°C for 12 hours, after ultrasonic cleaning with single distilled water, it is dried for later use;

[0047] B. Surface amination

[0048] The alkali-activated sample in step A was immersed in 2.5mg / ml polylysine (PLL, MW 150-300KDa) at 4°C for 12h, washed with phosphate buffer and then used;

[0049] C, Preparation of Sema 4D / Heparin complex

[0050] Mix Sema 4D with a concentration of 50ng / ml and Heparin with a concentration of 5mg / ml in equal volumes, and react at 37°C for 1h;

[0051] D. Sema 4D / Heparin fixed

[0052] Soak the sample in step B in the complex prepared in step ...

Embodiment 2

[0054]A method for constructing a Sema 4D / Heparin microenvironment on the surface of a cardiovascular implant material, comprising the following steps:

[0055] A. Alkaline activation

[0056] After the pure titanium is polished and cleaned, it is activated with 4mol / L NaOH for 16 hours. After ultrasonic cleaning with single distilled water, it is immersed in double distilled water. It is reacted at 80°C for 12 hours. After ultrasonic cleaning with single distilled water, it is dried for later use;

[0057] B. Surface amination

[0058] The alkali-activated sample in step A was immersed in 2.5mg / ml polylysine (PLL, MW 150-300KDa) at 4°C for 12h, washed with phosphate buffer and then used;

[0059] C, Preparation of Sema 4D / Heparin complex

[0060] Mix Sema 4D with a concentration of 200ng / ml and Heparin with a concentration of 8mg / ml in equal volumes, and react at 37°C for 3h;

[0061] D. Sema 4D / Heparin fixed

[0062] Soak the sample in step B in the complex prepared in s...

Embodiment 3

[0064] A method for constructing a Sema 4D / Heparin microenvironment on the surface of a cardiovascular implant material, comprising the following steps:

[0065] A. Alkaline activation

[0066] After the pure titanium is polished and cleaned, it is activated with 2mol / L NaOH for 10 hours, after ultrasonic cleaning with single distilled water, it is immersed in double distilled water, reacted at 80°C for 12 hours, after ultrasonic cleaning with single distilled water, it is dried for later use;

[0067] B. Surface amination

[0068] The alkali-activated sample in step A was immersed in 2.5mg / ml polylysine (PLL, MW 150-300KDa) at 4°C for 12h, washed with phosphate buffer and then used;

[0069] C, Preparation of Sema 4D / Heparin complex

[0070] Mix Sema 4D with a concentration of 400ng / ml and Heparin with a concentration of 10mg / ml in equal volumes, and react at 37°C for 2h;

[0071] D. Sema 4D / Heparin fixed

[0072] Soak the sample in step B in the complex prepared in step ...

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Abstract

The invention discloses a method for constructing a Sema 4D / Heparin microenvironment on the surface of a cardiovascular implanted material. The method comprises the following steps: A, performing alkali activation on a material by adopting NaOH; B, performing surface amino functionalization on the sample prepared in the step A; C, preparing a Heparin / Sema 4D compound; and D, fixing the Heparin / Sema 4D compound on the surface of the sample prepared in the step B. According to the method, Sema 4D and heparin are combined and together fixed on a material surface, so that Sema 4D can be protected and is prevented from being degraded by protease, identification and combination of Sema 4D and corresponding receptors can be promoted, and functions of the Sema 4D can be achieved. The method can be used for well simulating a body blood injury repairing process, has the advantages of simple process, is easy to operate, does not need expensive complicated equipment, and is applicable to surfaces of various complicated cardiovascular implanted appliances and other devices with requirement for resisting coagulation / promoting endothelial cell migration.

Description

technical field [0001] The invention relates to an inorganic material surface modification technology, in particular to a method for constructing a Sema 4D / Heparin microenvironment on the surface of a cardiovascular implant material. Background technique [0002] Cardiovascular system disease is one of the important diseases that endanger human health. Artificial implantation of vascular stents or heart valves is an effective way to treat cardiovascular diseases; currently, more than 2 million coronary heart disease patients worldwide need percutaneous coronary angioplasty every year In 2006, more than 100,000 patients received vascular stent implantation in my country; valvular heart disease accounts for about 30% of heart disease patients in China, and more than 100,000 critically ill patients must undergo manual surgery every year. Heart valve replacement. [0003] Titanium and its alloys are widely used in the field of biomedicine due to their good biocompatibility; vascu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/50A61L27/34A61L31/14A61L31/10
CPCA61L27/34A61L27/507A61L31/10A61L31/14A61L2400/18A61L2420/02A61L2420/06C08L89/00C08L5/10
Inventor 陈俊英崔园园周丰曾峥谭建英魏来牟小辉黄楠
Owner SOUTHWEST JIAOTONG UNIV
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