阅读说明：本技术 一种具有抗凝血和抗钙化功能的生物瓣膜及其制备方法 (Biological valve with anticoagulation and anti-calcification functions and preparation method thereof ) 是由 王云兵 李高参 杨立 于 2020-05-28 设计创作，主要内容包括：本发明公开了一种具有抗凝血和抗钙化功能的生物瓣膜及其制备方法。该方法的过程为：采用戊二醛对生物瓣膜进行交联处理,并在交联后采用亲水分子或疏水分子进行改性处理；或采用亲水分子或疏水分子对生物瓣膜进行改性处理,然后再进行戊二醛交联处理；或采用亲水分子或疏水分子对生物瓣膜进行至少一次改性处理,然后进行戊二醛交联处理,最后再用亲水分子或疏水分子对生物瓣膜进行至少一次改性处理。本发明制备得到的生物瓣膜不仅具有良好的机械性能,同时还具有优异的抗凝血和抗钙化效果。(The invention discloses a biological valve with anticoagulation and anti-calcification functions and a preparation method thereof. The method comprises the following steps: performing cross-linking treatment on the biological valve by using glutaraldehyde, and performing modification treatment by using hydrophilic molecules or hydrophobic molecules after cross-linking; or hydrophilic molecules or hydrophobic molecules are adopted to carry out modification treatment on the biological valve, and then glutaraldehyde crosslinking treatment is carried out; or hydrophilic molecules or hydrophobic molecules are adopted to carry out at least one modification treatment on the biological valve, then glutaraldehyde crosslinking treatment is carried out, and finally hydrophilic molecules or hydrophobic molecules are adopted to carry out at least one modification treatment on the biological valve. The biological valve prepared by the invention not only has good mechanical property, but also has excellent anticoagulation and anti-calcification effects.)

1. A preparation method of biological valve with anticoagulation and anti-calcification functions is characterized in that glutaraldehyde is adopted to carry out cross-linking treatment on the biological valve, and hydrophilic molecules or hydrophobic molecules are adopted to carry out modification treatment after cross-linking;

or hydrophilic molecules or hydrophobic molecules are adopted to carry out modification treatment on the biological valve, and then glutaraldehyde crosslinking treatment is carried out;

or hydrophilic molecules or hydrophobic molecules are adopted to carry out at least one modification treatment on the biological valve, then glutaraldehyde crosslinking treatment is carried out, and finally hydrophilic molecules or hydrophobic molecules are adopted to carry out at least one modification treatment on the biological valve.

2. The method for preparing a biological valve with anticoagulant and anticalcification functions as claimed in claim 1, wherein the specific process of the modification treatment is as follows:

and (3) placing the biological valve in a solution obtained by mixing hydrophilic molecules or hydrophobic molecules with a condensing agent, and reacting for 1-7 days at 4-37 ℃.

3. The method for preparing the biological valve with the anticoagulation and anti-calcification functions as claimed in claim 2, wherein the mass concentration of hydrophilic molecules in the mixed solution is 0.5-30%; the mass concentration of the hydrophobic molecules is 0.5-30%; the mass concentration of the condensing agent is 0.1-20%.

4. The method for preparing a biological valve with anticoagulant and anticalcification functions as claimed in any one of claims 1 to 3, wherein the hydrophilic molecule is at least one of taurines, glucosamine, aminoglycans, polylysine-polymethacryloxyethyl phosphocholine, polylysine-polymethacrylates, glycoproteins, short-chain polypeptides and phospholipids.

5. The method for preparing a biological valve with anticoagulant and anticalcification functions as in any one of claims 1 to 3, wherein the hydrophobic functional molecule comprises at least one of perfluorooctanoic acid-2-aminoethyl ester, (2-aminoethyl) -perfluorooctanoic amide, (2-aminoethyl) -heptadecafluoroundecanamide, perfluorododecylamine, perfluorooctylamine, perfluorohexylamine, perfluorobutylamine, and chitosan.

6. The method for preparing a biological valve with anticoagulation and anti-calcification functions as claimed in claim 1, wherein when glutaraldehyde is adopted for crosslinking treatment, the volume concentration of glutaraldehyde is 0.1-10%, and the pH value is 7-8.

7. The method for preparing a biological valve with anticoagulation and anti-calcification functions as claimed in claim 6, wherein the cross-linking reaction temperature is 4-37 ℃ and the reaction time is 1-7 days.

8. The method for preparing a biological valve with anticoagulant and anticalcification functions as claimed in claim 1, wherein before the glutaraldehyde crosslinking or modifying treatment, a cleaning pretreatment is further performed on the biological valve; the cleaning pretreatment process comprises the following steps: and (3) placing the biological valve in a rotating speed oscillation condition of 3-5 ℃ and 80-120 rpm, and cleaning for 1.5-3 h by using deionized water.

9. The method for preparing a biological valve with anticoagulant and anticalcification functions as claimed in claim 1, wherein the biological valve is a pulmonary valve, aortic valve, venous valve, mitral valve or tricuspid valve replacement and repair material.

10. The biological valve with the functions of anticoagulation and calcification prepared by the method of any one of claims 1-9.

Technical Field

The invention belongs to the technical field of preparation of medical materials and medical instruments, and particularly relates to a biological valve with anticoagulation and calcification-resisting functions and a preparation method thereof.

Background

Pulmonary valve diseases are mainly caused by pulmonary artery reflux and pulmonary artery stenosis, and the traditional surgical thoracotomy treatment is easy to cause serious pulmonary artery reflux, so that the right ventricular cavity is enlarged and the heart rate is abnormal. Further treatment requires another chest-opening operation, but the second (or more) operations have high operation difficulty, high risk and high death rate. In recent years, transcatheter interventional heart valve replacement techniques have brought about patient patency with the development of minimally invasive interventional techniques. The intervention heart valve replacement operation does not need to open the chest, the operation risk is low, the recovery time is short, and old patients and patients who are not suitable for opening the chest provide new treatment schemes.

Compared with the aortic valve, the pulmonary valve has the characteristics of slow pulmonary blood flow speed and generally low age in the range of applied patients, so that higher requirements are provided for the antithrombotic and anticalcification and durability performances of the interventional pulmonary valve. The problem of blood coagulation and calcification of the traditional glutaraldehyde cross-linked biological valve is a very important reason for the clinical failure of the valve. Therefore, the anticoagulation and anti-calcification treatment on the existing biological valve material can effectively prolong the service life of the interventional pulmonary valve, and has important significance for clinical use of the interventional pulmonary valve and improvement of the life quality of patients.

At present, a common hydrophilic and hydrophobic modification treatment method for glutaraldehyde valves is to modify hydrophilic or hydrophobic compounds on the surface of a valve material by using an electrostatic adsorption assembly mode. Because the hydrophilic and hydrophobic substances are modified on the surface of the valve material through weak binding force such as physical adsorption, the retention time of the hydrophilic and hydrophobic substances on the surface of the material is short, the performance is not stable, and the long-term anticoagulation and calcification resistance of the valve can not be realized.

Disclosure of Invention

Aiming at the defects in the prior art, the invention provides a biological valve with anticoagulation and anti-calcification functions and a preparation method thereof, aiming at solving the problems of coagulation and calcification of the existing interventional pulmonary valve, improving the anticoagulation and anti-calcification effects of a valve material and prolonging the service life of the valve while ensuring the mechanical performance of the biological valve material.

In order to achieve the purpose, the technical scheme adopted by the invention for solving the technical problems is as follows:

a preparation method of biological valve with anticoagulation and anti-calcification functions comprises the steps of carrying out cross-linking treatment on the biological valve by using glutaraldehyde, and carrying out modification treatment by using hydrophilic molecules or hydrophobic molecules after cross-linking;

or hydrophilic molecules or hydrophobic molecules are adopted to carry out modification treatment on the biological valve, and then glutaraldehyde crosslinking treatment is carried out;

or hydrophilic molecules or hydrophobic molecules are adopted to carry out at least one modification treatment on the biological valve, then glutaraldehyde crosslinking treatment is carried out, and finally hydrophilic molecules or hydrophobic molecules are adopted to carry out at least one modification treatment on the biological valve.

Further, the specific process of the modification treatment is as follows:

and (3) placing the biological valve in a solution obtained by mixing hydrophilic molecules or hydrophobic molecules with a condensing agent, and reacting for 1-7 days at 4-37 ℃.

Further, when modification treatment is required to be performed before and after glutaraldehyde crosslinking, the processes of the before and after modification treatment are independent of each other.

Further, the biological valve is placed in a mixed solution of the hydrophobic molecules and the condensing agent, soaked for 5 days at room temperature, then placed in a glutaraldehyde solution, soaked for 7 days at 37 ℃, and finally placed in a mixed solution of the hydrophobic molecules and the condensing agent, and soaked for 5 days at room temperature.

Further, the mass concentration of hydrophilic molecules in the mixed solution is 0.5-30%; the mass concentration of the condensing agent is 0.1-20%.

Further, the mass concentration of the hydrophilic molecules in the mixed solution is 0.5%, 1%, 8%, 15%, 20%, 30%, etc.; the mass concentration of the hydrophobic molecules is 0.5%, 1%, 8%, 15%, 20%, 30% and the like; the mass concentration of the condensing agent is 0.1%, 1%, 5%, 10%, 15%, etc.

Further, the hydrophilic molecule is at least one of taurate, glucosamine, aminoglycan, aminoglycerol, polylysine-polymethacryloxyethylphosphatidine, polylysine-polymethacrylate, glycoprotein, short chain polypeptide, and phospholipid.

Further, the hydrophobic functional molecule comprises at least one of perfluorooctanoic acid-2-aminoethyl ester, (2-aminoethyl) -perfluorooctanoic acid amide, (2-aminoethyl) -heptadecafluoroundecanamide, perfluorododecylamine, perfluorooctylamine, perfluorohexylamine, perfluorobutylamine, and chitosan.

Further, when glutaraldehyde is adopted for cross-linking treatment, the volume concentration of the glutaraldehyde is 0.1-10%, and the pH value is 7-8.

Furthermore, the crosslinking reaction temperature is 4-37 ℃, and the reaction time is 1-7 days.

Further, the volume concentration of glutaraldehyde is 0.1%, 1%, 5%, 10%, etc.; the pH value is 7, 7.4, 8 and the like.

Further, before the glutaraldehyde crosslinking or modification treatment, the biological valve is cleaned and pretreated; the cleaning process may include: fresh porcine or bovine pericardial tissue was collected and stored in a low temperature humidified state at 4 ℃. The pericardial tissue was washed with deionized (distilled) water using gentle friction and fluid pressure under shaking conditions to remove adherent non-pericardial and non-collagenous tissue. The washing described in the present invention achieves effective decellularization of pericardial tissue by osmotic shock, preferably washing continues until there is no visible adherent non-pericardial or non-collagenous tissue. In specific implementation, preferably, the step of cleaning the animal pericardium includes: cleaning fresh animal pericardium with deionized water for 1.5-3 h at 3-5 ℃ and at the rotating speed oscillation condition of 80-120 RPM. For example, the clean fresh animal (pig or cattle) pericardium can be subjected to a cross-linking treatment or a hydrophilic treatment after being washed with deionized water at 4 ℃ for 2 hours under a rotating speed oscillation condition of 100 RPM.

Further, the biological valve is a pulmonary valve, aortic valve, venous valve, mitral valve, or tricuspid valve replacement and repair material.

The biological valve with anticoagulation and anti-calcification functions prepared by the method.

The invention has the beneficial effects that:

1. the biological valve is placed in a mixed solution of hydrophilic molecules or hydrophobic molecules and a condensing agent, so that the hydrophilic molecules or hydrophobic molecules are fixed on the biological valve in a chemical grafting manner, and the biological valve has a good anticoagulation function. Meanwhile, the modification treatment can also seal valve calcification sites such as carboxyl, aldehyde group and the like of the glutaraldehyde valve, and simultaneously improve the long-acting anticoagulation and calcification resistance of the glutaraldehyde valve, thereby further improving the durability of biological valves, particularly pulmonary valve, and meeting the clinical requirement of the pulmonary valve.

2. According to the method, the biological valve is placed in a mixed solution of hydrophilic molecules or hydrophobic molecules and a condensing agent, then grafting reaction is carried out for 1-7 days at 4-37 ℃, hydrophilic molecules or hydrophobic molecules can be grafted on the biological valve as much as possible, and then stable crosslinking of most collagen tissues on the biological valve can be realized through crosslinking treatment of glutaraldehyde, so that the structural stability of the whole pericardial tissue is improved, and the immunogenicity is reduced or even eliminated.

3. The biological valve prepared by the method has good mechanical property, shows excellent anticoagulation and anti-calcification effects in tests such as coagulation tests and animal experiments, has excellent mechanical property and biocompatibility, can effectively reduce the coagulation and calcification of the biological valve, and prolongs the service life of the valve.

Drawings

FIG. 1 shows the results of mechanical testing of a biological valve prepared in example 1 of the present application;

FIG. 2 shows the results of a platelet adhesion test between a glutaraldehyde valve (left) and a biological valve prepared in example 1 of the present application (right);

fig. 3 shows the results of quantitative in vivo calcification analysis of glutaraldehyde valves (left) and biological valves prepared in example 1 of the present application (right).

Detailed Description

The following description of the embodiments of the present invention is provided to facilitate the understanding of the present invention by those skilled in the art, but it should be understood that the present invention is not limited to the scope of the embodiments, and it will be apparent to those skilled in the art that various changes may be made without departing from the spirit and scope of the invention as defined and defined in the appended claims, and all matters produced by the invention using the inventive concept are protected.