CN108785748B

CN108785748B - Multifunctional cardiovascular coating material with super-hydrophilic performance and preparation method thereof

Info

Publication number: CN108785748B
Application number: CN201810731279.9A
Authority: CN
Inventors: 王云兵; 李林华; 罗日方; 杨立; 庄伟华
Original assignee: Sichuan University
Current assignee: Sichuan University
Priority date: 2018-07-05
Filing date: 2018-07-05
Publication date: 2020-03-10
Anticipated expiration: 2038-07-05
Also published as: CN108785748A

Abstract

The invention discloses a multifunctional cardiovascular coating material with super-hydrophilic property and a preparation method thereof. The method comprises the following steps: (1) pretreating a substrate material; (2) placing the pretreated substrate material in a buffer system, adding a polyphenol compound and dopamine, and reacting for 2 hours at 4-50 ℃; adding sodium periodate, and reacting at 4-50 ℃; (3) and (3) ultrasonically cleaning the product obtained in the step (2) for 3-5 times by using deionized water, and drying by using nitrogen to obtain the multifunctional cardiovascular coating material. The multifunctional cardiovascular coating prepared by the invention has good stability, excellent hydrophilicity, good anticoagulation, anti-inflammation and anti-oxidation, capability of avoiding endothelial cell apoptosis and inhibiting smooth muscle cell proliferation, and can be used for preparing blood contact materials such as vascular stents, heart valves, artificial blood vessels, blood contact catheters and the like in the field of medical materials.

Description

Multifunctional cardiovascular coating material with super-hydrophilic performance and preparation method thereof

Technical Field

The invention belongs to the technical field of medical materials, and particularly relates to a multifunctional cardiovascular coating material with super-hydrophilic property and a preparation method thereof.

Background

Having excellent blood compatibility and cell compatibility is one of the conditions that biological materials must satisfy, and particularly for cardiovascular materials that are in direct contact with blood (e.g., vascular stents, heart valves, vascular prostheses, pacemakers, vascular prostheses, blood contact catheters, etc.), excellent blood compatibility is a primary consideration. After the material is contacted with blood, the adsorption and denaturation of plasma protein (such as fibrinogen and the like) occur in a very short time, and then the adhesion, activation and aggregation of platelets are accompanied, the formation of fibrin network is carried out, and finally red blood cells are netted to form thrombus aggregates. Finally, the implantation or the use of the device fails, so that the adsorption and the denaturation of the fibrinogen on the surface of the material are resisted, and the adsorption, the aggregation and the activation of the platelets on the surface of the biological material are inhibited, and the method for improving the blood compatibility of the biological material is one of the methods.

Meanwhile, after the material is implanted, an acute inflammatory reaction can be caused, the implant is identified by macrophages in an immune system through nonspecific adsorption of protein on the surface of the implant in a human body, and the macrophages form giant cells through self-fusion under the condition of not phagocytizing and secrete cytokines (reach a peak value within 24 hours), so that fibroblasts are promoted to be deposited on the surface of the implant to finally form a compact fibrous capsule and cover the implant, and the service of the implant material is adversely affected. The implant material has good blood compatibility, can effectively inhibit the activation and aggregation of inflammatory cells, and can effectively prevent the implant failure caused by the infection of blood contacting with instruments; the instrument can promote the growth of endothelial cells and inhibit the proliferation and migration of smooth muscle cells, further improve the blood compatibility of long-term blood contact materials and effectively avoid the implantation failure of the instrument caused by tissue hyperplasia. Therefore, the material has good anticoagulation, anti-inflammation, endothelial cell promotion and smooth muscle cell inhibition and is important for cardiovascular implantation materials.

For biomedical materials, the hydrophilicity/hydrophobicity of the material surface is the primary factor affecting protein adsorption. Meanwhile, because the protein is polyelectrolyte with amphoteric charges, if the surface of the material is also provided with an amphiphilic ion structure or a hydrophilic group, the interaction between the material and the protein can be weakened through enrichment of a hydration layer or spatial repulsion, and the adsorption of nonspecific protein is inhibited.

Moreover, in the current study on the influence of the hydrophilicity/hydrophobicity of the surface of the most widely used material on protein adsorption and cell compatibility, it is considered that the hydrophilic material has better biocompatibility than the hydrophobic material. Because the protein on the hydrophobic surface is firmly adhered, the protein is not easy to fall off and the conformational change of the protein is easy to cause; the hydrophilic surface has low adhesion strength to the protein, and is beneficial to the adjustment and maintenance of the original free conformation of the protein.

In the conventional method for preparing a hydrophilic surface, a hydrophilic functional group (such as a carboxylic acid group, a sulfonic acid group, an amine group, a quaternary amine group) or a substance with hydrophilicity is generally grafted or coated on the surface of a material, and whether the hydrophilic functional group is coated on the surface of the material by covalent fixation or physical blending, the hydrophilic functional group must depend on the size, conformation and activity of molecules and the stability and durability of the hydrophilic function on the surface of the material. Once the biomolecules are inactivated or largely lost in the service process, the biological functions of the biomolecules cannot be effectively exerted. Therefore, it is important to generate a large amount of stable hydrophilic groups directly on the surface of the coating layer and not be affected by the grafting process.

Disclosure of Invention

Aiming at the defects in the prior art, the invention provides a multifunctional cardiovascular coating material with super-hydrophilic property and a preparation method thereof, and the multifunctional cardiovascular coating material with anticoagulation, anti-inflammation, antioxidation, endothelial cell apoptosis prevention and smooth muscle cell proliferation inhibition effects can be prepared.

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 multifunctional cardiovascular coating material with super-hydrophilic performance comprises the following steps:

(1) pretreating a substrate material;

(2) placing the pretreated substrate material in a buffer system with the pH value of 3-6, and then adding a polyphenol compound with the final concentration of 0.01-10 mg/mL and dopamine with the final concentration of 0.01-10 mg/mL to react for 2 hours at the temperature of 4-50 ℃; adding sodium periodate with the final concentration of 0.01mg/mL-10mg/mL, and reacting at the temperature of 4-50 ℃ for 0.1-48 h;

(3) and (3) ultrasonically cleaning the product obtained in the step (2) by using deionized water for 3-5 times, and drying by using nitrogen for 2-5 min each time to obtain the multifunctional cardiovascular coating material with super-hydrophilic performance.

Further, the substrate material in the step (1) is a metal material, a ceramic material, a polymer material or a composite biological material.

Wherein the metal material is stainless steel, cobalt-based alloy, titanium and alloy thereof, magnesium and alloy thereof or pure iron;

the ceramic material is medical inorganic material and film-TiO₂Low temperature Pyrolytic Carbon (LTIC), glassy Carbon, diamond-like Carbon, or alumina (Al)₂O₃)；

The high polymer material is polyethylene, polyvinyl chloride, polytetrafluoroethylene, polypropylene, polymethyl methacrylate, medical polysulfone, medical polysiloxane, polyether sulfone membrane, polylactic acid, polyglycolic acid, glycolide-lactide copolymer PLGA, polyurethane, medical collagen or natural rubber;

the composite biological material is stainless steel fiber, titanium fiber reinforced bioglass, polylactic acid, glycolic acid fiber or hydroxyapatite reinforced polylactic acid.

Further, the final concentration of the polyphenol compound in the step (2) was 2 mg/mL.

Further, the polyphenol compound is tannic acid, gallic acid, salvianolic acid B, epigallocatechin gallate, epicatechin, epigallocatechin, catechol, pyrogallol, flavonoid, anthocyanin, ellagic acid or proanthocyanidin A2.

Further, the final concentration of dopamine in step (2) was 4 mg/mL.

Further, in the step (2), the final concentration of sodium periodate was 4 mg/ml.

Further, in the step (2), the buffer system 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, citric acid-sodium hydroxide-hydrochloric acid buffer solution or citric acid-sodium citrate buffer solution.

The multifunctional cardiovascular coating material with super-hydrophilic performance prepared by the method.

The invention has the beneficial effects that:

1. polyphenol compounds have been extensively studied in the field of adhesion chemistry because of their ability to interact with substrate materials due to their ortho-phenolic hydroxyl functionality. In addition, polyphenols can be chemically cross-linked with compounds containing amino or thiol groups and can be deposited on the surface of various metals, inorganic materials and polymers to form polymeric films containing ortho-phenolic groups. Meanwhile, the polyphenol compound has good antioxidant capacity, the performance of inhibiting the growth of bacteria, the performance of inhibiting the activation and aggregation of macrophages, the performance of strengthening blood vessel walls, the performance of promoting the growth of endothelial cells and the performance of inhibiting the proliferation and migration of smooth muscle cells. Therefore, in recent years, polyphenol compounds have been widely studied and paid attention to in the field of surface modification of biomedical materials.

2. The polyphenol compound, the dopamine and the sodium periodate interact with the surface of the material in various modes such as covalent bond, hydrogen bond, hydrophobic acting force, supermolecule acting force and the like to generate physical/chemical deposition to form a polyphenol coating; in addition, in the reaction process of the polyphenol compound and the oxidant, the polyphenol compound is attacked by electrons of the oxidant, so that an o-diquinone structure is converted into a carboxyl structure, a large amount of carboxyl is generated on the coating, a hydrophilic surface is formed, and various hydrophilic functional groups (such as-COOH and-NH) are generated on the surface of the coating₂phenol-OH, etc.), thereby enabling the coating to have good hydrophilicity, benefiting from the good hydrophilicity, enabling the surface of the coating to have good stain resistance, and effectively resisting non-specific proteins (such as: fibrinogen, albumin, etc.) and further inhibits platelet adhesion and activation.

3. In addition, the unreacted polyphenols in the coating can exert their good biological functions, including antioxidant, anti-inflammatory, antibacterial, endothelial cell friendly and smooth muscle cell proliferation inhibition, etc. In addition, a large number of hydrophilic groups can be used as a platform for subsequent biomolecule grafting modification, so that the biocompatibility of the surface of the material can be effectively improved. At present, no report that the coating which has the functions of resisting inflammation and blood coagulation, avoiding apoptosis of inner cells, inhibiting proliferation of smooth muscle cells and is rich in active functional groups is prepared on the surface of a biological material by utilizing the action of polyphenol and an oxidizing agent is reported.

4. The molecular structure of the polyphenol compound contains a large amount of ortho-phenolic hydroxyl groups, the ortho-phenolic hydroxyl groups can form stable chelation with metal in a coating, the polyphenol cross-linked coating can form stable combination on the surface of the metal, the cross-linked polymeric coating can also form stable combination with a substrate material through intermolecular hydrophobic interaction and hydrogen bond interaction, and the coating has excellent binding force with various biomaterial substrates, and is particularly suitable for preparing blood contact materials such as vascular stents, heart valves, artificial blood vessels, blood contact catheters and the like.

5. The coating required to be prepared by the invention is usually less than 200nm, the obtained coating is uniform, the raw material investment required for preparing the coating is small, the raw material investment amount is easy to regulate and control, and the coating can be modified on the surfaces of various materials without influencing the performance of the body material. Compared with the traditional anticoagulation coating technology, the method has the advantages of simple operation, lower cost and wide universality.

Drawings

FIG. 1 is a Scanning Electron Microscope (SEM) image of platelet adhesion; wherein A is a Scanning Electron Microscope (SEM) image of the platelet adhesion of the traditional polyphenol deposited film; b is a Scanning Electron Microscope (SEM) image of the platelet adhesion of the multifunctional cardiovascular coating obtained in example 3;

FIG. 2 is a diagram of laser Confocal (CLSM) of smooth muscle cell adhesion; wherein, A is the laser Confocal (CLSM) picture of the adhesion of the smooth muscle cells of the traditional polyphenol deposited thin film, and B is the laser Confocal (CLSM) picture of the adhesion of the smooth muscle cells of the multifunctional cardiovascular coating obtained in the embodiment 3.

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

(1) polishing the medical stainless steel to be modified and modified, and then cleaning and drying;

(2) placing the treated stainless steel in an acetic acid-acetate buffer solution with the pH value of 4, and then sequentially adding gallic acid with the final concentration of 0.01mg/mL and dopamine with the final concentration of 0.01mg/mL in a buffer system to react for 2h at 40 ℃; then adding sodium periodate solution with final concentration of 0.01mg/ml, and reacting for 30h at 40 ℃;

(3) and (3) ultrasonically cleaning the product obtained in the step (2) by deionized water for 5min each time, and drying under the condition of nitrogen to prepare the multifunctional cardiovascular coating which has super-hydrophilic performance, anticoagulation, anti-inflammation and antioxidation, avoids endothelial cell apoptosis and inhibits smooth muscle cell proliferation.

Example 2

(1) polishing the nickel-titanium alloy to be modified and modified, and then cleaning and drying;

(2) placing the treated stainless steel in a phthalic acid-hydrochloric acid buffer solution with the pH value of 3.5, then adding epigallocatechin gallate (EGCG) with the final concentration of 1mg/mL and dopamine with the final concentration of 2mg/mL in a buffer system, and reacting for 2h at 30 ℃; then adding sodium periodate solution with the final concentration of 2mg/mL, and reacting for 10h at 30 ℃;

(3) and (3) washing the product obtained in the step (2) with deionized water for 5min each time, and drying under the condition of nitrogen to obtain the multifunctional cardiovascular coating which has super-hydrophilic performance, anticoagulation, anti-inflammation and anti-oxidation, can avoid endothelial cell apoptosis and can inhibit smooth muscle cell proliferation.

Example 3

(1) cleaning and drying the polytetrafluoroethylene needing modification;

(2) placing the treated stainless steel in a 2- (N-morpholine) ethanesulfonic acid buffer solution with the pH value of 6, and then sequentially adding epicatechin gallate (ECG) with the final concentration of 2mg/mL and dopamine with the final concentration of 4mg/mL in a buffer system to react for 2h at 15 ℃; then adding sodium periodate solution with the final concentration of 4mg/mL, and reacting for 24h at 15 ℃;

Example 4

(1) cleaning and drying the polyurethane to be modified and modified;

(2) placing the treated stainless steel in a citric acid-sodium citrate buffer solution with the pH value of 6, and then sequentially adding gallic acid with the final concentration of 2mg/mL and dopamine with the final concentration of 5mg/mL in a buffer system to react for 2h at the temperature of 20 ℃; then adding sodium periodate solution with the final concentration of 2mg/mL, and reacting for 18h at 20 ℃;

Example 5

(1) polishing the isotropic pyrolytic carbon LTIC material to be modified and modified, and then cleaning and drying;

(2) placing the treated stainless steel in a potassium hydrogen phthalate-sodium hydroxide buffer solution with the pH value of 4.5, and then sequentially adding a compound with Epicatechin (EC) with the final concentration of 5mg/mL and dopamine with the final concentration of 7mg/mL in a buffer system to react for 2h at 25 ℃; then adding sodium periodate solution with final concentration of 7mg/ml, and reacting for 6h at 25 ℃;

Example 6

(1) cleaning and drying the medical collagen needing to be modified and modified;

(2) placing the treated stainless steel in a glycine-hydrochloric acid buffer solution with the pH value of 4, and then sequentially adding epigallocatechin EGC with the final concentration of 0.1mg/mL and dopamine with the final concentration of 0.5mg/mL in a buffer system to react for 2 hours at 30 ℃; then adding sodium periodate solution with final concentration of 0.5mg/mL, and reacting for 12h at 30 ℃;

Example 7

(1) cleaning and drying the natural rubber needing modification;

(2) placing the treated stainless steel in a disodium hydrogen phosphate-citric acid buffer solution with the pH value of 5.5, and then sequentially adding pyrogallol with the final concentration of 7mg/mL and dopamine with the final concentration of 10mg/mL in a buffer system to react for 2h at 40 ℃; then adding sodium periodate solution with the final concentration of 10mg/mL, and reacting for 1h at 40 ℃;

Example 8

(1) cleaning and drying the hydroxyapatite reinforced polylactic acid to be modified and modified;

(2) placing the treated stainless steel in an acetic acid-acetate buffer solution with the pH value of 6, and then sequentially adding gallic acid with the final concentration of 3mg/mL and dopamine with the final concentration of 1mg/mL in a buffer system to react for 2h at 45 ℃; then adding sodium periodate solution with the final concentration of 1mg/mL, and reacting for 4h at 45 ℃;

Detection of

The adhesion of platelets and smooth muscle cells of the conventional polyphenol coating and the multifunctional cardiovascular coating prepared in example 3 of the present invention were examined, respectively, and the results are shown in fig. 1 and fig. 2, respectively.

As shown in fig. 1, wherein a is a Scanning Electron Microscope (SEM) image of platelet adhesion of a traditional polyphenol deposited film, and B is a Scanning Electron Microscope (SEM) image of platelet adhesion of the multifunctional cardiovascular coating obtained in example 3; as can be seen from the detection result graph, compared with the conventional polyphenol coating, the platelet adhesion quantity, aggregation and activation degree of the surface of the multifunctional cardiovascular coating prepared in the embodiment 3 of the present invention are significantly inhibited, which indicates that the multifunctional cardiovascular coating prepared in the present invention has excellent anti-platelet adhesion performance.

As shown in fig. 2, wherein a is the laser Confocal (CLSM) map of the adhesion of smooth muscle cells of the conventional polyphenol deposited thin film, and B is the laser Confocal (CLSM) map of the adhesion of smooth muscle cells of the multifunctional cardiovascular coating obtained in example 3; as can be seen from the detection result graph, compared with the traditional polyphenol coating, the adhesion number and the growth form of the smooth muscle cells on the surface of the multifunctional cardiovascular coating prepared in the embodiment 3 of the invention are significantly inhibited, which indicates that the multifunctional cardiovascular coating prepared in the invention has excellent performance of inhibiting smooth muscle cell proliferation.

Claims

1. A preparation method of a multifunctional cardiovascular coating material with super-hydrophilic performance is characterized by comprising the following steps:

(1) pretreating a substrate material;

(3) and (3) ultrasonically cleaning the product obtained in the step (2) by using deionized water for 3-5 times, wherein each time lasts for 2-5 min, and drying by using nitrogen to obtain the multifunctional cardiovascular coating material.

2. The method for preparing multifunctional cardiovascular coating material with super hydrophilic property as claimed in claim 1, wherein the base material in step (1) is metal material, ceramic material, polymer material or composite biomaterial.

3. The method for preparing multifunctional cardiovascular coating material with super hydrophilic property as claimed in claim 1, wherein the final concentration of polyphenol compound in step (2) is 2 mg/mL.

4. The method for preparing multifunctional cardiovascular coating material with super hydrophilic property according to claim 1 or 3, wherein the polyphenol compound is tannic acid, gallic acid, salvianolic acid B, epigallocatechin gallate, epicatechin, epigallocatechin, catechol, pyrogallol, flavonoid, anthocyanin, ellagic acid or proanthocyanidin A2.

5. The method for preparing multifunctional cardiovascular coating material with super hydrophilic property according to claim 1, wherein the final concentration of dopamine in step (2) is 4 mg/mL.

6. The method for preparing multifunctional cardiovascular coating material with super hydrophilic property as claimed in claim 1, wherein the final concentration of sodium periodate in step (2) is 4 mg/ml.

7. The method for preparing a multifunctional cardiovascular coating material with super-hydrophilic property according to claim 1, wherein the buffer system in step (2) is acetic acid-acetate buffer, 2- (N-morpholine) ethanesulfonic acid buffer, glycine-hydrochloric acid buffer, phthalic acid-hydrochloric acid buffer, potassium hydrogen phthalate-sodium hydroxide buffer, disodium hydrogen phosphate-citric acid buffer, citric acid-sodium hydroxide-hydrochloric acid buffer or citric acid-sodium citrate buffer.

8. The multifunctional cardiovascular coating material with super-hydrophilic property prepared by the method of any one of claims 1 to 7.