CN108785748A - A kind of Multifunctional heart blood vessel coating material and preparation method thereof with super hydrophilicity - Google Patents

Abstract

The invention discloses a multifunctional cardiovascular coating material with superhydrophilic performance and a preparation method thereof. The method comprises the following steps: (1) pretreating the base material; (2) placing the pretreated base material in a buffer system, then adding polyphenol compounds and dopamine, and reacting at 4-50° C. for 2 hours; Then add sodium periodate and react at 4-50 DEG C; (3) ultrasonically clean the product obtained in step (2) with deionized water for 3-5 times, dry with nitrogen gas, and become a multi-functional cardiovascular coating material. The multifunctional cardiovascular coating prepared by the present invention has good stability, excellent hydrophilicity, good anticoagulation, anti-inflammation, anti-oxidation, ability to avoid endothelial cell apoptosis and inhibit smooth muscle cell proliferation , can be used in the preparation of blood-contact materials such as vascular stents, heart valves, artificial blood vessels, and blood-contact catheters in the field of medical materials.

Description

A multifunctional cardiovascular coating material with superhydrophilic properties and its preparation method

technical field

The invention belongs to the technical field of medical materials, and in particular relates to a multifunctional cardiovascular coating material with superhydrophilic performance and a preparation method thereof. Preparation of contact materials.

Background technique

Having excellent blood compatibility and cell compatibility is one of the conditions that biological materials must meet, especially for cardiovascular materials that are in direct contact with blood (such as: vascular stents, heart valves, artificial blood vessels, pacemakers, artificial Blood vessels, blood contact catheters, etc.), excellent blood compatibility is the primary consideration. After the material comes into contact with blood, the adsorption and denaturation of plasma proteins (such as fibrinogen, etc.) occurs in a very short period of time, followed by the adhesion, activation and aggregation of platelets, the formation of fibrin network, and finally the formation of thrombus by recruiting red blood cells Aggregates. It will eventually lead to the failure of device implantation or use. Therefore, preventing the adsorption and denaturation of fibrinogen on the surface of materials, thereby inhibiting the adsorption, aggregation and activation of platelets on the surface of biomaterials, is one of the methods to improve the hemocompatibility of biomaterials.

At the same time, the implantation of the material will cause an acute inflammatory reaction, and the non-specific adsorption of proteins on the surface of the implant in the human body will make the implant recognized by the macrophages in the immune system, and the macrophages will pass through the body if the phagocytosis fails. Self-fusion to form giant cells and secrete cytokines (peak within 24 hours), prompting fibroblasts to deposit on the surface of the implant and eventually form a dense fibrous capsule and cover the implant, thereby serving the implant material bring adverse effects. The implant material has good blood compatibility, and can effectively inhibit the activation and aggregation of inflammatory cells, which can effectively prevent the implantation failure caused by the infection of the blood contact device; the device can promote the growth of endothelial cells and inhibit the smooth muscle cells Proliferation and migration performance can further improve the blood compatibility of long-term blood contact materials and effectively avoid device implantation failure caused by tissue proliferation. Therefore, it is very important for cardiovascular implant materials to have better anticoagulation, anti-inflammation, promotion of endothelial cells, and inhibition of smooth muscle cell growth.

For biomedical materials, the affinity/hydrophobicity of the material surface is the primary factor affecting protein adsorption. At the same time, since proteins are polyelectrolytes with amphoteric charges, if the surface of the material also has amphiphilic ionic structures or hydrophilic groups, the interaction between the material and the protein can also be weakened by enriching the hydration layer or steric repulsion, inhibiting the Adsorption of non-specific proteins.

Moreover, among the effects of hydrophilic/hydrophobic properties on the surface of materials that are most widely studied at present on protein adsorption and cell compatibility, it is generally agreed that hydrophilic materials have better biocompatibility than hydrophobic materials. Because the hydrophobic surface protein adheres more firmly, it is not easy to fall off, and it is easy to cause the conformational change of the protein; while the adhesion strength of the hydrophilic surface to the protein is low, which is conducive to the adjustment and maintenance of the original free conformation of the protein.

The traditional method of preparing a hydrophilic surface is generally to graft or coat a hydrophilic functional group (such as a carboxylic acid group, a sulfonic acid group, an amine group, a quaternary ammonium group) or a hydrophilic substance on the material. The surface, whether it is covalently immobilized or physically blended to coat the surface of the material, must depend on the size, conformation, activity of the molecule, and the stability and persistence of its hydrophilic effect on the surface of the material. Once biomolecules are inactivated or lost in large quantities during service, they cannot effectively perform their biological functions. Therefore, it is particularly important to generate a large number of stable hydrophilic groups directly on the coating surface without being affected by the grafting process.

Contents of the invention

Aiming at the above-mentioned deficiencies in the prior art, the present invention provides a multi-functional cardiovascular coating material with superhydrophilic properties and a preparation method thereof, which can prepare a coating material with anticoagulation, anti-inflammation, anti-oxidation, anti-endothelial A multifunctional cardiovascular coating that inhibits apoptosis and proliferation of smooth muscle cells.

In order to achieve the above object, the technical solution adopted by the present invention to solve the technical problems is:

A preparation method of a multifunctional cardiovascular coating material with superhydrophilic properties, comprising the following steps:

(1) Carry out pretreatment to substrate material;

(2) Place the pretreated base material in a buffer system with a pH value of 3 to 6, then add polyphenol compounds with a final concentration of 0.01 mg/mL to 10 mg/mL and a final concentration of 0.01 mg/mL to 10mg/mL dopamine was reacted at 4-50°C for 2h; then sodium periodate with a final concentration of 0.01mg/mL-10mg/mL was added, and reacted at 4-50°C for 0.1-48h;

(3) Ultrasonic cleaning of the product obtained in step (2) with deionized water for 3 to 5 times, each time for 2 to 5 minutes, and drying with nitrogen gas to obtain a multifunctional cardiovascular coating material with superhydrophilic properties.

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

Among them, the metal materials are stainless steel, cobalt-based alloys, titanium and its alloys, magnesium and its alloys, or pure iron;

Ceramic materials are medical inorganic materials and thin films-TiO ₂ , Low Temperature Isotropic Pyrolytic Carbon (LTIC), glassy carbon, diamond, diamond-like carbon or aluminum oxide (Al ₂ O ₃ );

Polymer materials are polyethylene, polyvinyl chloride, polytetrafluoroethylene, polypropylene, polymethyl methacrylate, medical polysulfone, medical polysiloxane, polyethersulfone membrane, polylactic acid, polyglycolic acid, glycolic acid Ester-lactide copolymer PLGA, polyurethane, medical collagen or natural rubber;

The composite biomaterial 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 step (2) is 2 mg/mL.

Further, the polyphenolic compound is tannic acid, gallic acid, salvianolic acid B, epigallocatechin gallate, epicatechin gallate, epicatechin, epigallocatechin, phthalate Phenols, pyrogallol, flavonoids, anthocyanins, ellagic acid or proanthocyanidins A2.

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

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

Further, 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, o-phthalic acid Potassium hydrogen diformate-sodium hydroxide buffer, disodium hydrogen phosphate-citric acid buffer, citric acid-sodium hydroxide-hydrochloric acid buffer or citric acid-sodium citrate buffer.

The multifunctional cardiovascular coating material with superhydrophilic properties prepared by the above method.

The beneficial effects of the present invention are:

1. Polyphenolic compounds have been widely studied in the field of adhesion chemistry because of their ortho-phenolic hydroxyl functional groups and the interaction with substrate materials. In addition, polyphenols can be chemically cross-linked with compounds containing amino or mercapto groups, and can be deposited on the surface of various metals, inorganic materials and polymers to form polymeric films containing ortho-phenolic groups. At the same time, polyphenol compounds have good antioxidant capacity, inhibit the growth of bacteria, inhibit the activation and aggregation of macrophages, strengthen the blood vessel wall, promote the growth of endothelial cells, and inhibit the proliferation and migration of smooth muscle cells. Therefore, in recent years, polyphenolic compounds have also received extensive research and attention in the field of surface modification of biomedical materials.

2. Polyphenolic compounds, dopamine and sodium periodate interact in multiple modes such as covalent, hydrogen bond, hydrophobic force and supramolecular force, and physically/chemically deposit with the surface of the material to form a polyphenol coating and, during the reaction of the polyphenol compound with the oxidizing agent, it is attacked by the electrons of the oxidizing agent, so that the o-diquinone structure is converted into a carboxyl structure, so that the coating has a large amount of carboxyl groups to form a hydrophilic surface, and a variety of Hydrophilic functional groups (such as -COOH, -NH ₂ , phenol-OH, etc.), so that the coating has good hydrophilicity, thanks to the excellent hydrophilicity, the surface of the coating has good anti-fouling properties It can effectively resist the adhesion and denaturation of non-specific proteins (such as fibrinogen, albumin, etc.), and further prevent the adhesion and activation of platelets.

3. In addition, the unreacted polyphenols in the coating can exert its good biological functions, including antioxidant, anti-inflammatory, antibacterial, friendly to endothelial cells and inhibiting smooth muscle cell proliferation, etc. In addition, a large number of hydrophilic groups can be used as a platform for subsequent biomolecular graft modification, which can effectively improve the biocompatibility of the material surface. At present, there is no report on the use of polyphenols and oxidants to prepare coatings on the surface of biomaterials that are anti-inflammatory, anti-coagulant, avoid inner cell apoptosis, inhibit smooth muscle cell proliferation, and are rich in active functional groups.

4. The molecular structure of polyphenolic compounds contains a large number of ortho phenolic hydroxyl groups. In the coating, these ortho phenolic hydroxyl groups can form a stable chelation with metals, and the polyphenol crosslinked coating can form a relatively Stable combination, the cross-linked polymeric coating can also form a relatively stable combination with the substrate material through intermolecular hydrophobic interactions and hydrogen bonds, such a coating has excellent binding force with various biological material substrates, especially suitable for Preparation of blood-contact materials such as vascular stents, heart valves, artificial blood vessels, and blood-contact catheters.

5. The coating required by the present invention is usually lower than 200nm, and the resulting coating is uniform. The raw material input required for preparing the coating is small, and the amount of raw material input is easy to control. The coating can be modified on the surface of various materials without affecting properties of the bulk material. Compared with the traditional anti-coagulation coating technology, the operation is simple, the cost is low, and the universality is wide.

Description of drawings

Fig. 1 is the scanning electron microscope (SEM) figure of platelet adhesion; Wherein, A is the scanning electron microscope (SEM) figure of traditional polyphenol deposition film platelet adhesion; Scanning electron microscope (SEM) picture;

Fig. 2 is the laser confocal (CLSM) figure of smooth muscle cell adhesion; Wherein, A is the laser confocal (CLSM) figure of smooth muscle cell adhesion of traditional polyphenol deposition film, B is the multifunctional cardiovascular coating obtained in embodiment 3 Confocal laser (CLSM) images of smooth muscle cell adhesion.

Detailed ways

The specific embodiments of the present invention are described below so that those skilled in the art can understand the present invention, but it should be clear that the present invention is not limited to the scope of the specific embodiments. For those of ordinary skill in the art, as long as various changes Within the spirit and scope of the present invention defined and determined by the appended claims, these changes are obvious, and all inventions and creations using the concept of the present invention are included in the protection list.

Example 1

A preparation method of a multifunctional cardiovascular coating material with superhydrophilic properties, comprising the following steps:

(1) Polish the medical stainless steel that needs to be modified, and then clean and dry;

(2) Place the treated stainless steel in an acetic acid-acetate buffer solution with a pH value of 4, then add gallic acid with a final concentration of 0.01 mg/mL and gallic acid with a final concentration of 0.01 mg/mL in the buffer system successively. Dopamine was reacted at 40°C for 2h; then sodium periodate solution with a final concentration of 0.01mg/ml was added, and reacted at 40°C for 30h;

(3) Use deionized water to ultrasonically clean the product obtained in step (2) for 3 times, each time for 5 minutes, and then dry it under nitrogen to prepare a super-hydrophilic, anti-coagulant, anti-inflammatory and anti-inflammatory product. Oxidation, avoidance of endothelial cell apoptosis, inhibition of smooth muscle cell proliferation in a multifunctional cardiovascular coating.

Example 2

A preparation method of a multifunctional cardiovascular coating material with superhydrophilic properties, comprising the following steps:

(1) Polish the nickel-titanium alloy that needs to be modified, and then clean and dry;

(2) Place the treated stainless steel in a phthalic acid-hydrochloric acid buffer solution with a pH value of 3.5, and then add epigallocatechin gallate (EGCG) with a final concentration of 1 mg/mL in the buffer system, Dopamine with a final concentration of 2mg/mL was reacted at 30°C for 2h; then sodium periodate solution with a final concentration of 2mg/mL was added and reacted at 30°C for 10h;

(3) Wash the product obtained in step (2) with deionized water for 3 times, each time for 5 minutes, and then dry it under nitrogen to prepare a super-hydrophilic, anti-coagulant, anti-inflammatory, and anti-oxidative product , Avoid endothelial cell apoptosis, inhibit smooth muscle cell proliferation multifunctional cardiovascular coating.

Example 3

A preparation method of a multifunctional cardiovascular coating material with superhydrophilic properties, comprising the following steps:

(1) cleaning and drying the polytetrafluoroethylene that needs to be modified;

(2) Place the treated stainless steel in 2-(N-morpholine) ethanesulfonic acid buffer solution with a pH value of 6, and then add epicatechin gallic acid with a final concentration of 2 mg/mL in the buffer system Ester (ECG) and dopamine with a final concentration of 4mg/mL were reacted at 15°C for 2h; then sodium periodate solution with a final concentration of 4mg/mL was added and reacted at 15°C for 24h;

(3) Wash the product obtained in step (2) with deionized water for 3 times, each time for 5 minutes, and then dry it under nitrogen to prepare a super-hydrophilic, anti-coagulant, anti-inflammatory, and anti-oxidative product , Avoid endothelial cell apoptosis, inhibit smooth muscle cell proliferation multifunctional cardiovascular coating.

Example 4

A preparation method of a multifunctional cardiovascular coating material with superhydrophilic properties, comprising the following steps:

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

(2) Place the treated stainless steel in a citric acid-sodium citrate buffer solution with a pH value of 6, then add gallic acid with a final concentration of 2 mg/mL and dopamine with a final concentration of 5 mg/mL in the buffer system React at 20°C for 2h; then add sodium periodate solution with a final concentration of 2mg/mL, and react at 20°C for 18h;

(3) Wash the product obtained in step (2) with deionized water for 3 times, each time for 5 minutes, and then dry it under nitrogen to prepare a super-hydrophilic, anti-coagulant, anti-inflammatory, and anti-oxidative product , Avoid endothelial cell apoptosis, inhibit smooth muscle cell proliferation multifunctional cardiovascular coating.

Example 5

A preparation method of a multifunctional cardiovascular coating material with superhydrophilic properties, comprising the following steps:

(1) Polish the isotropic pyrolytic carbon LTIC material that needs to be modified, then clean and dry;

(2) Place the treated stainless steel in a potassium hydrogen phthalate-sodium hydroxide buffer solution with a pH value of 4.5, and then sequentially add epicatechin (EC) with a final concentration of 5 mg/mL in the buffer system. ) compound and dopamine with a final concentration of 7mg/mL were reacted at 25°C for 2h; then sodium periodate solution with a final concentration of 7mg/ml was added and reacted at 25°C for 6h;

(3) Wash the product obtained in step (2) with deionized water for 3 times, each time for 5 minutes, and then dry it under nitrogen to prepare a super-hydrophilic, anti-coagulant, anti-inflammatory, and anti-oxidative product , Avoid endothelial cell apoptosis, inhibit smooth muscle cell proliferation multifunctional cardiovascular coating.

Example 6

A preparation method of a multifunctional cardiovascular coating material with superhydrophilic properties, comprising the following steps:

(1) Clean and dry the medical collagen that needs to be modified;

(2) Place the treated stainless steel in a glycine-hydrochloric acid buffer solution with a pH value of 4, then add epigallocatechin EGC with a final concentration of 0.1 mg/mL in the buffer system, and a final concentration of 0.5 mg/mL. mL of dopamine was reacted at 30°C for 2h; then sodium periodate solution with a final concentration of 0.5mg/mL was added and reacted at 30°C for 12h;

(3) Wash the product obtained in step (2) with deionized water for 3 times, each time for 5 minutes, and then dry it under nitrogen to prepare a super-hydrophilic, anti-coagulant, anti-inflammatory, and anti-oxidative product , Avoid endothelial cell apoptosis, inhibit smooth muscle cell proliferation multifunctional cardiovascular coating.

Example 7

A preparation method of a multifunctional cardiovascular coating material with superhydrophilic properties, comprising the following steps:

(1) cleaning and drying the natural rubber that needs to be modified;

(2) Place the treated stainless steel in a disodium hydrogen phosphate-citric acid buffer solution with a pH value of 5.5, then add successively pyrogallol with a final concentration of 7 mg/mL in the buffer system, and a final concentration of 10 mg/mL. mL of dopamine was reacted at 40°C for 2h; then sodium periodate solution with a final concentration of 10mg/mL was added and reacted at 40°C for 1h;

(3) Wash the product obtained in step (2) with deionized water for 3 times, each time for 5 minutes, and then dry it under nitrogen to prepare a super-hydrophilic, anti-coagulant, anti-inflammatory, and anti-oxidative product , Avoid endothelial cell apoptosis, inhibit smooth muscle cell proliferation multifunctional cardiovascular coating.

Example 8

A preparation method of a multifunctional cardiovascular coating material with superhydrophilic properties, comprising the following steps:

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

(2) Place the treated stainless steel in the acetic acid-acetate buffer solution with a pH value of 6, then add gallic acid with a final concentration of 3 mg/mL and dopamine with a final concentration of 1 mg/mL in the buffer system successively. React at 45°C for 2h; then add sodium periodate solution with a final concentration of 1mg/mL, and react at 45°C for 4h;

(3) Wash the product obtained in step (2) with deionized water for 3 times, each time for 5 minutes, and then dry it under nitrogen to prepare a super-hydrophilic, anti-coagulant, anti-inflammatory, and anti-oxidative product , Avoid endothelial cell apoptosis, inhibit smooth muscle cell proliferation multifunctional cardiovascular coating.

detection

The adhesion of platelets and smooth muscle cells of the traditional polyphenol coating and the multifunctional cardiovascular coating prepared in Example 3 of the present invention were tested respectively, and the results are shown in Fig. 1 and Fig. 2 respectively.

As shown in Figure 1, wherein, the scanning electron microscope (SEM) figure of A traditional polyphenol deposition film platelet adhesion, B is the scanning electron microscope (SEM) figure of embodiment 3 gained multifunctional cardiovascular coating platelet adhesion; From detection As can be seen from the result figure, compared with the traditional polyphenol coating, the platelet adhesion quantity, aggregation and activation degree on the surface of the multifunctional cardiovascular coating prepared in Example 3 of the present invention are significantly inhibited, indicating that the present invention prepares The multifunctional cardiovascular coating has excellent anti-platelet adhesion properties.

As shown in Figure 2, wherein, A is the laser confocal (CLSM) figure of traditional polyphenol deposition film smooth muscle cell adhesion, and B is the laser confocal (CLSM) of embodiment 3 multifunctional cardiovascular coating smooth muscle cell adhesion ) figure; as can be seen from the detection result figure, compared with the traditional polyphenol coating, the adhesion quantity and growth form of the smooth muscle cells on the surface of the multifunctional cardiovascular coating prepared in Example 3 of the present invention are significantly inhibited, It shows that the multifunctional cardiovascular coating prepared by the present invention has excellent performance of inhibiting the proliferation of smooth muscle cells.

Claims (8)

1. a preparation method of a multifunctional cardiovascular coating material with superhydrophilic properties, is characterized in that, comprises the following steps: (1) Carry out pretreatment to substrate material; (2) Place the pretreated base material in a buffer system with a pH value of 3 to 6, then add polyphenol compounds with a final concentration of 0.01 mg/mL to 10 mg/mL and a final concentration of 0.01 mg/mL to 10mg/mL dopamine was reacted at 4-50°C for 2h; then sodium periodate with a final concentration of 0.01mg/mL-10mg/mL was added, and reacted at 4-50°C for 0.1-48h; (3) Ultrasonic cleaning of the product obtained in step (2) with deionized water for 3 to 5 times, each time for 2 to 5 minutes, and drying with nitrogen gas. It is a multifunctional cardiovascular coating material. 2. the preparation method of the multifunctional cardiovascular coating material with superhydrophilic property according to claim 1, is characterized in that, base material described in step (1) is metal material, ceramic material, polymer material or composite biomaterials. 3. the preparation method of the multifunctional cardiovascular coating material with superhydrophilic property according to claim 1, is characterized in that, the final concentration of polyphenol compound described in step (2) is 2mg/mL. 4. the preparation method of the multifunctional cardiovascular coating material with superhydrophilic property according to claim 1 or 3, is characterized in that, described polyphenolic compound is tannic acid, gallic acid, salvianolic acid B, Epigallocatechin gallate, epicatechin gallate, epicatechin, epigallocatechin, catechol, pyrogallol, flavonoids, anthocyanins, ellagic acid, or proto Anthocyanin A2. 5. the preparation method of the multifunctional cardiovascular coating material with superhydrophilic property according to claim 1, is characterized in that, the final concentration of dopamine described in step (2) is 4mg/mL. 6. the preparation method of the multifunctional cardiovascular coating material with superhydrophilic property according to claim 1, is characterized in that, the final concentration of sodium periodate described in step (2) is 4mg/ml. 7. the preparation method of the multifunctional cardiovascular coating material with superhydrophilic property according to claim 1, is characterized in that, buffer system described in step (2) is acetic acid-acetate damping fluid, 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 superhydrophilic properties prepared by the method according to any one of claims 1 to 7.