阅读说明：本技术 一种长效超疏水抗凝生物瓣膜及其制备方法 (Long-acting super-hydrophobic anticoagulation biological valve and preparation method thereof ) 是由 杨立 王云兵 罗日方 于 2020-12-22 设计创作，主要内容包括：本发明公开了一种长效超疏水抗凝生物瓣膜及其制备方法。该方法包括以下步骤：(1)采用戊二醛处理生物瓣膜材料48～96h；(2)在生物瓣膜材料表面嵌入由多酚化合物和金属离子聚合而成,且含有双键基团的纳米颗粒；(3)在纳米颗粒表面接枝疏水性物质形成疏水涂层即可。本发明根据多酚温和的自聚合反应条件和金属离子、双键与巯基反应机理启发,在不影响瓣膜本体性能的同时,通过简单、稳定的操作工艺即在生物瓣膜表面制备具有长效的高水接触角和低滚动角的超疏水涂层,通过阻抗血浆蛋白的吸附实现长效抗凝血需求。(The invention discloses a long-acting super-hydrophobic anticoagulation biological valve and a preparation method thereof. The method comprises the following steps: (1) treating the biological valve material with glutaraldehyde for 48-96 hours; (2) embedding nanoparticles which are formed by polymerization of polyphenol compounds and metal ions and contain double-bond groups on the surface of a biological valve material; (3) and grafting a hydrophobic substance on the surface of the nano-particles to form a hydrophobic coating. According to the invention, inspired by the mild self-polymerization reaction condition of polyphenol and the reaction mechanism of metal ions, double bonds and sulfydryl, the long-acting super-hydrophobic coating with high water contact angle and low rolling angle is prepared on the surface of the biological valve by a simple and stable operation process without influencing the performance of the valve body, and the long-acting anticoagulation requirement is realized by resisting the adsorption of plasma protein.)

1. A preparation method of a long-acting super-hydrophobic anticoagulation biological valve is characterized by comprising the following steps:

(1) treating a biological valve material by using glutaraldehyde;

(2) embedding nanoparticles which are formed by polymerization of polyphenol compounds and metal ions and contain double-bond groups on the surface of a biological valve material;

(3) and grafting a hydrophobic substance on the surface of the nano-particles to form a hydrophobic coating.

2. The method for preparing a long-acting superhydrophobic anticoagulant biological valve according to claim 1, wherein the specific process of the step (2) is as follows:

and (2) placing the biological valve material treated in the step (1) into acid liquor containing polyphenol compounds and metal ions, adding an oxidant with the concentration of 20 mu M-1 mM, and reacting at 10-40 ℃ for 20-120 min.

3. The preparation method of the long-acting super-hydrophobic anticoagulant biological valve according to claim 1 or 2, wherein the molar mass ratio of the polyphenol compound to the metal ions is 1: 0.01-1.

4. The method of claim 3, wherein the polyphenol compound is at least one of tannic acid, gallic acid, salvianolic acid B, epigallocatechin gallate, epicatechin, epigallocatechin, catechol, pyrogallol, and flavones.

5. The method of claim 3, wherein the metal ions are at least one of copper ions, silver ions and iron ions.

6. The method of claim 2, wherein the acid solution is an acetate buffer, a 2- (N-morpholine) ethanesulfonic acid buffer, a glycine-hydrochloric acid buffer, a phthalic acid-hydrochloric acid buffer, a potassium hydrogen phthalate-sodium hydroxide buffer, a disodium hydrogen phosphate-citric acid buffer, or a citric acid-sodium hydroxide-hydrochloric acid buffer.

7. The method for preparing a long-acting superhydrophobic anticoagulant biological valve according to claim 2, wherein the oxidant is a water-soluble oxidant, specifically at least one of hydrogen peroxide, ammonium persulfate, copper chloride, ferric trichloride, concentrated nitric acid, sodium periodate, potassium permanganate, and potassium dichromate.

8. The method for preparing a long-acting superhydrophobic anticoagulant biological valve according to claim 1, wherein the specific process of the step (3) is as follows:

and (3) placing the product obtained in the step (2) into a hydrophobic substance with the concentration of 50 mu M-1 mM for soaking for 10-30 min, and forming a hydrophobic coating on the surface of the nano-particles by a click chemistry method.

9. The method for preparing a long-acting superhydrophobic anticoagulation biological valve according to claim 8, wherein the hydrophobic substance is perfluoropentadecane, perfluorododecanoic acid, 1H,2H, 2H-perfluorododecanethiol, perfluorobutylethylene, octadecanoyl chloride or stearic acid.

10. The method for preparing a long-acting superhydrophobic anticoagulant biological valve according to claim 1, wherein the biological valve material is an aortic valve, a pulmonary valve, a venous valve, a mitral valve, or a tricuspid valve.

Technical Field

The invention belongs to the technical field of valve material preparation, and particularly relates to a long-acting super-hydrophobic anticoagulation biological valve and a preparation method thereof.

Background

With the aging stage of the population structure of China, patients with severe valvular stenosis, regurgitation and the like are increased obviously earlier, the overall incidence rate of people over 75 years old is high, once the patients have symptoms and are not treated, the death rate is close to 50% within 2 to 3 years. Minimally invasive interventional valvuloplasty is an important treatment measure accepted by more and more patients due to small operation trauma, low risk and good effect, and a plurality of corresponding problems are met along with the deep development of the minimally invasive interventional technology. Wherein implantation of the biological valve in interventional therapy can lead to thrombosis, and thrombus on the valve surface falls off to increase the risk of postoperative stroke; and thrombosis on the surface of the valve can seriously affect the durability of the valve. Therefore, the biological valve material has excellent long-acting anticoagulation effect and is crucial to the durability of the valve and the life safety of patients.

The super-hydrophobic surface of the bionic lotus leaf structure is widely researched due to the high water contact angle (150 degrees) and the low rolling angle, and is mainly applied to the fields of self-cleaning, moisture prevention, water-oil separation, corrosion resistance, pollution resistance and the like. Once the surface of a common biological material is contacted with blood or body fluid, a large amount of protein or cells can be adhered to the surface of the material, and the performance of the material is seriously influenced; whereas both superhydrophilic and superhydrophobic surfaces have good anti-fouling (impedance to non-specific adhesion of proteins, cells and microorganisms) effects. Studies have reported that superhydrophobic surfaces have lower platelet adhesion capacity than superhydrophilic surfaces, indicating that superhydrophobic surfaces have superior anti-thrombotic properties. The existing super-hydrophobic surface is mainly prepared on the surfaces of metal, polymer and inorganic materials, and the research on preparing the super-hydrophobic coating on the surface of the biological valve material is less, so that the preparation of the super-hydrophobic coating with high water contact angle and low adhesion force on the surface of the biological valve material is important for resisting the thrombosis after the valve is implanted into a body.

Disclosure of Invention

Aiming at the defects in the prior art, the invention provides a long-acting super-hydrophobic anticoagulation biological valve and a preparation method thereof, which can prepare a layer of super-hydrophobic coating with high water contact angle and low rolling angle on the surface of a valve material under the condition of not changing the mechanical property of a valve body, thereby improving the antithrombotic property of the biological valve after being implanted into a body.

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 a long-acting super-hydrophobic anticoagulation biological valve comprises the following steps:

(1) treating the biological valve material with glutaraldehyde for 48-96 hours;

(2) embedding nanoparticles which are formed by polymerization of polyphenol compounds and metal ions and contain double-bond groups on the surface of a biological valve material;

(3) and grafting a hydrophobic substance on the surface of the nano-particles to form a hydrophobic coating.

Further, the specific process of the step (2) is as follows:

and (2) placing the biological valve material treated in the step (1) into acid liquor containing polyphenol compounds and metal ions, adding an oxidant with the concentration of 20 mu M-1 mM, and reacting at 10-40 ℃ for 20-120 min.

Furthermore, the molar mass ratio of the polyphenol compound to the metal ions is 1: 0.01-1.

Further, the polyphenol compound is at least one of tannic acid, gallic acid, salvianolic acid B, epigallocatechin gallate, epicatechin, epigallocatechin, catechol, pyrogallol and flavone.

Further, the metal ions are at least one of copper ions, silver ions, and iron ions.

Further, the pH value of the acid liquor is 4-6.

Further, the acid solution is acetic acid-acetate buffer solution, 2- (N-morpholine) ethanesulfonic acid buffer solution, glycine-hydrochloric acid buffer solution, phthalic acid-hydrochloric acid buffer solution, potassium hydrogen phthalate-sodium hydroxide buffer solution, disodium hydrogen phosphate-citric acid buffer solution or citric acid-sodium hydroxide-hydrochloric acid buffer solution.

Further, the oxidant is a water-soluble oxidant, specifically at least one of hydrogen peroxide, ammonium persulfate, cupric chloride, ferric trichloride, concentrated nitric acid, sodium periodate, potassium permanganate and potassium dichromate.

Further, the specific process of the step (3) is as follows:

and (3) placing the product obtained in the step (2) into a hydrophobic substance with the concentration of 50 mu M-1 mM for soaking for 10-30 min, and forming a hydrophobic coating on the surface of the nano-particles by a click chemistry method.

Further, the hydrophobic substance is perfluoropentadecane, perfluorododecanoic acid, 1H,2H, 2H-perfluorododecanethiol, perfluorobutylethylene, octadecanoyl chloride or stearic acid.

Further, the biological valve material is an aortic valve, a pulmonary valve, a venous valve, a mitral valve, or a tricuspid valve.

Further, the source of the biological valve material is porcine pericardium, bovine pericardium and the like.

The invention has the beneficial effects that:

1. according to the invention, the polyphenol compound can be introduced by utilizing the residual amino and aldehyde groups on the surface of the valve material and through the reactivity of polyphenol related functional groups, in addition, the nanoparticle product obtained by oxidizing the polyphenol compound and metal in the presence of a strong oxidant is rich in double-bond groups, and the biological valve material with long-acting super-hydrophobic anticoagulation performance can be prepared by subsequently introducing hydrophobic fluoride with functional group modification through click chemistry. The preparation method has the advantages of simple and controllable preparation process, mild conditions and stable super-hydrophobic property of the coating, and can effectively resist platelet adhesion and thrombosis on the surface of the valve.

2. According to the invention, inspired by the mild self-polymerization reaction condition of polyphenol and the reaction mechanism of metal ions, double bonds and sulfydryl, the long-acting super-hydrophobic coating with high water contact angle and low rolling angle is prepared on the surface of the biological valve by a simple and stable operation process without influencing the performance of the valve body, and the long-acting anticoagulation requirement is realized by resisting the adsorption of plasma protein.

Drawings

FIG. 1 is an electron microscope image of the surface of a super-hydrophobic fluoride coating of a biological valve prepared by an example;

FIG. 2 is a contact angle result for the coating of FIG. 1;

FIG. 3 is the in vitro platelet adhesion results of a conventional glutaraldehyde valve;

fig. 4 shows the in vitro platelet adhesion results of the superhydrophobic modified valve material prepared in example 2 of the invention.

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.

Example 1

A preparation method of a long-acting super-hydrophobic anticoagulation biological valve comprises the following steps:

(1) pretreating the glutaraldehyde-crosslinked porcine pericardium valve material;

(2) immersing the porcine heart valve-wrapping material treated in the step (1) in a solution containing tannic acid and Cu in a molar mass ratio of 1:0.1²⁺Adding acetic acid-sodium acetate as buffer solution with pH of 4 into ionic buffer solution, adding sodium periodate oxidant with concentration of 20 μ M, and reacting at 25 deg.C for 40min to make tannin undergo oxidative polymerization reaction and simultaneously react with Cu²⁺Ion chelation to generate Cu-containing ions on the surface of the biological valve material²⁺The polyphenol nanoparticles of (a);

(3) the surface contains Cu²⁺The polyphenol nanoparticle porcine pericardium valve material is placed in 1H,1H,2H, 2H-perfluorododecanethiol with the molar concentration of 0.8mM to be soaked for 20min, and then a layer of uniform fluoride coating is generated on the surface of the valve material by a chemical grafting method, so that the biological valve material with the surface provided with the super-hydrophobic coating is prepared.

Example 2

A preparation method of a long-acting super-hydrophobic anticoagulation biological valve comprises the following steps:

(1) pretreating the glutaraldehyde-crosslinked porcine pericardium valve material;

(2) will be treated by the step (1)Immersing the porcine pericardium material in a solution containing epigallocatechin gallate and Ag at a molar mass ratio of 1:0.05⁺Adding glycine-hydrochloric acid buffer solution with pH of 4 into ionic buffer solution, adding 80 μ M concentrated nitric acid oxidant, reacting at 25 deg.C for 60min to allow epigallocatechin gallate to undergo oxidative polymerization reaction and simultaneously react with Ag⁺Ion chelation to generate Ag on the surface of the biological valve material⁺The polyphenol nanoparticles of (a);

(3) the surface contains Ag⁺The polyphenol nanoparticle porcine pericardium valve material is soaked in perfluorododecanoic acid with the molar concentration of 0.5mM for 25min, and then a layer of uniform and compact fluoride coating is generated on the surface of the valve material by a chemical grafting method, so that the biological valve material with the surface having the super-hydrophobic coating is prepared.

Example 3

A preparation method of a long-acting super-hydrophobic anticoagulation biological valve comprises the following steps:

(1) pretreating the glutaraldehyde-crosslinked bovine pericardial valve material;

(2) immersing the porcine pericardium material treated in the step (1) in a solution containing epicatechin and Ag in a molar mass ratio of 1:0.2⁺Adding disodium hydrogen phosphate-citric acid buffer solution with pH of 4 into ionic buffer solution, adding 0.05mM potassium permanganate oxidant into the solution, and reacting at 20 deg.C for 40min to allow epicatechin to undergo oxidative polymerization and Ag⁺Ion chelation to generate Ag on the surface of the biological valve material⁺The polyphenol nanoparticles of (a);

(3) the surface contains Ag⁺The polyphenol nano-particle bovine pericardial valve material is soaked in octadecanoyl chloride with the molar concentration of 0.2mM for 10min, and then a layer of uniform and compact hydrophobic coating is generated on the surface of the valve material by a chemical grafting method, so that the biological valve material with the super-hydrophobic coating on the surface is prepared.

Example 4

A preparation method of a long-acting super-hydrophobic anticoagulation biological valve comprises the following steps:

(1) pretreating the glutaraldehyde-crosslinked bovine pericardial valve material;

(2) immersing the porcine pericardium material treated in the step (1) in a solution containing pyrogallol and Fe with the molar mass ratio of 1:0.05³⁺Adding citric acid-sodium hydroxide-hydrochloric acid buffer solution into the ionic mixed solution, wherein the pH value of the buffer solution is 4, adding ammonium persulfate oxidant with the concentration of 0.01mM into the solution, and reacting at 20 ℃ for 60min to enable pyrogallol to perform oxidative polymerization reaction and simultaneously react with Fe³⁺Ion chelation to generate Fe-containing ions on the surface of the biological valve material³⁺The polyphenol nanoparticles of (a);

(3) the surface contains Fe³⁺The polyphenol nano-particle bovine pericardial valve material is soaked in stearic acid with the molar concentration of 0.05mM for 30min, and then a layer of uniform and compact hydrophobic coating is generated on the surface of the valve material by a chemical grafting method, so that the biological valve material with the surface having the super-hydrophobic coating is prepared.

Examples of the experiments

1. An electron microscope image of the hydrophobic coating of the biological valve material prepared in example 2 is shown in fig. 1, and a contact angle detection result is shown in fig. 2.

As shown in fig. 1 and fig. 2, the hydrophobic coating prepared by the present application is uniformly distributed on the surface of the valve material, and has long-acting high water contact angle and low rolling angle.

2. In-vitro platelet adhesion experiments are carried out by taking glutaraldehyde valves as a control group and the biological valve material prepared in example 2 as an experimental group, and the anticoagulation performance of the experiments is respectively detected, and the results are shown in fig. 3 and 4.

As shown in fig. 3 and 4, the biological valve material prepared by the method has an excellent anticoagulation effect compared with a glutaraldehyde valve.