A super-hydrophilic coating with anticoagulant and anti-inflammatory functions and its preparation method and application

Abstract

The present invention discloses a super-hydrophilic coating with anti-coagulation and anti-inflammatory functions, a preparation method and an application thereof, and belongs to the technical field of biomedical materials. The present invention first prepares a polyphenol solution, and then constructs a hydrophilic coating on the surface of a substrate; then, the polyphenol solution is mixed with a magnesium-containing solution, and a hydrophilic amino-containing macromolecule is added, and finally, the resulting product is coated on the surface of the hydrophilic substrate to obtain a super-hydrophilic coating with anti-coagulation and anti-inflammatory functions. The super-hydrophilic coating prepared by the present invention has anti-coagulation and anti-inflammatory properties, and also has a function of promoting endothelialization, which can significantly reduce the risk of thrombosis and the degree of inflammatory response after device implantation. The super-hydrophilic coating of the present invention can be applied to the surface modification of cardiovascular materials / devices such as vascular stents, heart occluders, artificial heart valves and artificial blood vessels.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedical materials, and in particular to a super-hydrophilic coating with anti-coagulation and anti-inflammatory functions, a preparation method and an application thereof. Background Art

[0002] Cardiovascular implantable devices, such as vascular stents, heart occluders, artificial heart valves and artificial blood vessels, are prone to poor blood compatibility and strong immune rejection reactions when implanted into the human body due to the lack of long-term effective anticoagulant and inflammation regulating surfaces, which can lead to a variety of clinical complications and failure of implantable devices. Surface modification of implantable devices or coating with effective anticoagulant and anti-inflammatory coatings is an important way to solve this problem. However, the weak adhesion between the coating and the surface of the implantable device and the poor stability of the coating make it difficult to function for a long time in the in vivo environment, which is the main problem limiting the application of the coating. Summary of the invention

[0003] In order to solve the above-mentioned deficiencies in the prior art, the purpose of the present invention is to provide a super hydrophilic coating with anticoagulant and anti-inflammatory functions and a preparation method and application thereof, so as to solve the problems of poor blood compatibility and strong immune rejection reaction of the existing coating.

[0004] The technical solution of the present invention to solve the above technical problems is as follows: a method for preparing a super-hydrophilic coating with anticoagulant and anti-inflammatory functions is provided, comprising the following steps:

[0005] (1) dissolving polyphenol compounds in a Tris-HCl solution to prepare a polyphenol solution;

[0006] (2) Cleaning and drying the surface of the substrate material, and then constructing a hydrophilic coating on the surface of the substrate material to obtain a hydrophilic substrate;

[0007] (3) The polyphenol compound solution of step (1) is mixed with a magnesium-containing solution, and then a hydrophilic amino-containing macromolecule is added, and then the resulting product is coated on the surface of the hydrophilic substrate obtained in step (2).

[0008] Furthermore, the polyphenol compound in step (1) is at least one of tannic acid, salvianolic acid B, gallic acid, epigallocatechin, epigallocatechin gallate, epicatechin gallate, catechol, pyrogallol, epicatechin, anthocyanin, flavonoids, ellagic acid and proanthocyanidin A2.

[0009] Furthermore, the concentration of the polyphenol solution in step (1) is 1-4 mg / mL.

[0010] Furthermore, in step (2), the substrate material is a metal-based biomaterial, a polymer-based biomaterial or a decellularized biological tissue.

[0011] Furthermore, in step (2), the water contact angle of the surface of the hydrophilic substrate is less than 60°.

[0012] Furthermore, in step (3), the hydrophilic amino-containing macromolecule is polyethyleneimine or polyallylamine, and the concentration of the hydrophilic amino-containing macromolecule is 0.1-1 mg / mL.

[0013] Furthermore, the magnesium-containing solution in step (3) is at least one of a magnesium sulfate solution, a magnesium chloride solution and a magnesium nitride solution.

[0014] Furthermore, the coating method in step (2) is drip coating, spray coating or dipping.

[0015] The invention provides a super-hydrophilic coating with anti-coagulation and anti-inflammatory functions, which is prepared by the above-mentioned preparation method.

[0016] The present invention also provides an application of a super-hydrophilic coating with anticoagulation and anti-inflammatory functions in the preparation of cardiovascular materials / devices.

[0017] Furthermore, the blood contact material / device is a vascular stent, a heart occluder, an artificial heart valve, an artificial blood vessel, a cardiopulmonary circulation circuit or an intravenous catheter for dialysis.

[0018] The present invention has the following beneficial effects:

[0019] (1) The present invention utilizes metal-phenol coordination to prepare a super-hydrophilic coating with a phenolic magnesium framework. The preparation method of the present invention can form a magnesium ion covering layer on the surface of a hydrophilic substrate material, which can effectively prevent calcium ions from occupying the surface of the substrate material, thereby inhibiting two coagulation pathways, including the intrinsic coagulation pathway and the extrinsic coagulation pathway. At the same time, the phenolic magnesium framework coating can cover the surface of the substrate material with magnesium ions for a long time by slowly releasing magnesium ions. Magnesium ions can promote the transformation of pro-inflammatory macrophages (M1) to anti-inflammatory macrophages (M2), greatly reducing the inflammatory risk of biomedical implantable materials.

[0020] (2) The preparation method of the super-hydrophilic coating of the present invention is simple and the conditions are mild. When polyphenols and magnesium ions form a complex in the solution, they can be evenly distributed on the surface of the substrate material through the ion effect in the solution to form a stable array structure, so that the coating has excellent stability and anti-fouling adhesion effect, and can be applied to the surface treatment of various biomedical materials.

[0021] (3) When preparing the super-hydrophilic coating, the present invention adds positively charged hydrophilic amino macromolecules, which can enhance the cross-linking effect between the coating components and improve the toughness of the coating.

[0022] (4) The super-hydrophilic coating with "dual-resistance" effect of the present invention can promote the endothelialization process on the surface of the substrate through polyphenols. At the same time, the magnesium ions on the surface of the substrate material can inhibit the growth of smooth muscle by regulating calcium ion channels, thereby achieving the function of rapid endothelialization and inhibition of smooth muscle growth of biomedical implantable interventional devices to prevent restenosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 The figure is a schematic diagram of the preparation process of the super-hydrophilic coating with anticoagulant and anti-inflammatory functions of the present invention;

[0024] Figure 2 The super-hydrophilic coating and the bare polylactic acid material prepared in Example 1 are scanning electron microscope images; wherein, Figure A is the bare polylactic acid material, and Figure B is the super-hydrophilic coating prepared in Example 1;

[0025] Figure 3 The water contact angle test diagrams of the super-hydrophilic coating prepared in Example 1 and the surface of the bare polylactic acid material; wherein, Figure A is the bare polylactic acid material, and Figure B is the super-hydrophilic coating prepared in Example 1;

[0026] Figure 4 The scanning electron microscope images of platelet adhesion of the super-hydrophilic coating and the bare polylactic acid material prepared in Example 1; wherein, Figure A is the bare polylactic acid material, and Figure B is the super-hydrophilic coating prepared in Example 1;

[0027] Figure 5 The diagram shows the percentage of pro-inflammatory macrophages M1 and anti-inflammatory macrophages M2 in the super-hydrophilic coating and naked polylactic acid material prepared in Example 1; wherein, Figure A shows the naked polylactic acid material, and Figure B shows the super-hydrophilic coating prepared in Example 1. DETAILED DESCRIPTION

[0028] The following examples are only used to explain the present invention and are not intended to limit the scope of the present invention. If no specific conditions are specified in the examples, the conditions are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0029] The preparation process of the super-hydrophilic coating with anticoagulant and anti-inflammatory functions in Example 1-4 is as follows Figure 1 As shown, the specific process is as follows:

[0030] Example 1

[0031] A method for preparing a super-hydrophilic coating with anticoagulant and anti-inflammatory functions comprises the following steps:

[0032] (1) dissolving tannic acid in a Tris-HCl solution (50 mM, pH = 8.5) to prepare a tannic acid solution; wherein the concentration of the tannic acid solution is 2 mg / mL;

[0033] (2) Place the cleaned polylactic acid material in a Tris-HCl buffer solution (50 mM, pH = 8.5) containing 2 mg / mL dopamine hydrochloride until the solution covers the surface of the substrate, react at 30 ° C for 2 h, and wash with deionized water three times;

[0034] (3) Mix equal volumes of the tannic acid solution obtained in step (1) and a Tris-HCl solution (50 MM, pH = 8.5) containing 1 mg / mL magnesium chloride. Take 10 mL of the mixed solution, then add 100 µL of a 5 mg / mL polyethyleneimine solution. After thorough mixing, drop the mixture onto the surface of the polylactic acid material obtained in step (2) at a drop volume of 0.6 mL / cm 2 , react at room temperature for 15 minutes, repeat the dropwise addition 15 times, and then wash three times with deionized water to obtain.

[0035] Depend on Figure 2 It can be seen that the surface of the bare polylactic acid material is smooth, and the surface of the super hydrophilic coating prepared in this embodiment has complex nanoparticles formed by tannic acid and magnesium ions. The water contact angles of the bare polylactic acid material and the super hydrophilic coating prepared in this embodiment are shown in Figure 2. Figure 3 As shown by Figure 3 It can be seen that the super-hydrophilic coating prepared in this embodiment forms a super-hydrophilic interface.

[0036] Example 2

[0037] A method for preparing a super-hydrophilic coating with anticoagulant and anti-inflammatory functions comprises the following steps:

[0038] (1) dissolving tannic acid in a Tris-HCl solution (50 mM, pH = 8.5) to prepare a tannic acid solution; wherein the concentration of the tannic acid solution is 2 mg / mL;

[0039] (2) Place the cleaned polylactic acid material in a Tris-HCl buffer solution (50 mM, pH = 8.5) containing 2 mg / mL dopamine hydrochloride until the solution covers the surface of the substrate, react at 30 ° C for 2 h, and wash with deionized water three times;

[0040] (3) Mix the tannic acid solution obtained in step (1) with an equal volume of a Tris-HCl solution (50 mM, pH = 8.5) containing 1 mg / mL magnesium chloride, take 10 mL of the mixed solution, then add 100 µL of a 5 mg / mL polyethyleneimine solution, mix thoroughly, and then drop it onto the surface of the polylactic acid material obtained in step (2), with a drop volume of 0.6 mL / cm 2 , react at room temperature for 15 minutes, repeat the dropwise addition 15 times, and then wash three times with deionized water to obtain.

[0041] Example 3

[0042] A method for preparing a super-hydrophilic coating with anticoagulant and anti-inflammatory functions comprises the following steps:

[0043] (1) dissolving epigallocatechin gallate in a Tris-HCl solution (50 mM, pH = 8.5) to prepare an epigallocatechin gallate solution; wherein the concentration of the epigallocatechin gallate solution is 2 mg / mL;

[0044] (2) Place the cleaned titanium foil material in a Tris-HCl buffer solution (50 mM, pH = 8.5) containing 2 mg / mL dopamine hydrochloride until the solution covers the surface of the substrate, react at 30 ° C for 2 h, and wash with deionized water three times;

[0045] (3) Mix the epigallocatechin gallate solution obtained in step (1) and Tris-HCl solution (50 mM, pH = 8.5) containing 1 mg / mL magnesium sulfate in equal volumes, take 10 mL of the mixed solution, then add 100 µL of 5 mg / mL polyallylamine solution, mix thoroughly and then drop it onto the surface of the titanium foil obtained in step (2), with a drop volume of 0.6 mL / cm 2 , react at room temperature for 15 minutes, repeat the dropwise addition 15 times, and then wash three times with deionized water to obtain.

[0046] Example 4

[0047] A method for preparing a super-hydrophilic coating with anticoagulant and anti-inflammatory functions comprises the following steps:

[0048] (1) dissolving epigallocatechin gallate in a Tris-HCl solution (50 mM, pH = 8.5) to prepare an epigallocatechin gallate solution; wherein the concentration of the epigallocatechin gallate solution is 2 mg / mL;

[0049] (2) Place the cleaned titanium foil material in a Tris-HCl buffer solution (50 mM, pH = 8.5) containing 2 mg / mL dopamine hydrochloride until the solution covers the surface of the substrate, react at 30 ° C for 2 h, and wash with deionized water three times;

[0050] (3) The epigallocatechin gallate solution obtained in step (1) was mixed with an equal volume of a Tris-HCl solution (50 mM, pH = 8.5) containing 1 mg / mL magnesium sulfate, 10 mL of the mixed solution was taken, and then 100 µL of a 5 mg / mL polyallylamine solution was added. After thorough mixing, the mixture was injected into a gas-tight syringe, and the nozzle was adjusted to a distance of 5 cm from the surface of the titanium foil material. The solution was sprayed on the surface of the titanium foil material obtained in step (2) at an ultrasonic frequency of 120 kHz, an ultrasonic power of 1.5 W, and a gas pressure of 1 psi. The propulsion rate of the solution was 0.04 mL / min, and the amount of solution sprayed each time was controlled to be 0.6 mL / cm 2 Repeat the spraying 15 times, and then wash with deionized water three times to obtain.

[0051] Test Example 1 Platelet Adhesion Test

[0052] The super-hydrophilic coating and naked polylactic acid material prepared in Example 1 were mixed with platelet-rich plasma, respectively, and incubated at 37° C. for 2 h. The adhesion of platelets on the super-hydrophilic coating and naked polylactic acid material prepared in Example 1 was observed by scanning electron microscopy. Figure 4 .Depend on Figure 4 It can be seen that compared with the naked polylactic acid material, the platelets adhered to the surface of the superhydrophilic coating prepared in Example 1 are significantly reduced, so the superhydrophilic coating prepared in the present invention has an anti-coagulation function.

[0053] Test Example 2 Anti-inflammatory Test

[0054] The super-hydrophilic coating prepared in Example 1 and the naked polylactic acid material were co-cultured with macrophages for 3 days, and the proportions of pro-inflammatory macrophages M1 and anti-inflammatory macrophages M2 on the surfaces of the super-hydrophilic coating prepared in Example 1 and the naked polylactic acid material were observed. Figure 5 .

[0055] Depend on Figure 5 It can be seen that compared with the naked polylactic acid material, the proportion of anti-inflammatory macrophages on the surface of the superhydrophilic coating prepared in Example 1 is higher, so the superhydrophilic coating prepared in the present invention has anti-inflammatory function.

[0056] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing a super-hydrophilic coating having anticoagulant and anti-inflammatory functions, characterized in that: The following steps are involved: (1) dissolving polyphenol compounds in a Tris-HCl solution to prepare a polyphenol solution; (2) cleaning and drying the surface of the substrate material, and then placing it in a Tris-HCl buffer solution containing 2 mg / mL dopamine hydrochloride, and reacting it at 30° C. for 2 hours to obtain a hydrophilic substrate; wherein the water contact angle of the surface of the hydrophilic substrate is less than 60°; (3) mixing the polyphenol compound solution of step (1) with the magnesium-containing solution, then adding a hydrophilic amino-containing macromolecule, reacting at room temperature for 15 minutes, and then coating the resulting product on the surface of the hydrophilic substrate obtained in step (2); Wherein, the hydrophilic amino-containing macromolecule is polyethyleneimine or polyallylamine; and the concentration of the hydrophilic amino-containing macromolecule is 0.1-1 mg / mL.

2. The method for preparing a super-hydrophilic coating having anticoagulant and anti-inflammatory functions according to claim 1, characterized in that: The polyphenol compound in step (1) is at least one of tannic acid, salvianolic acid B, gallic acid, epigallocatechin, epigallocatechin gallate, epicatechin gallate, catechol, pyrogallol, epicatechin, anthocyanin, flavonoids, ellagic acid and proanthocyanidin A2.

3. The method for preparing a super-hydrophilic coating having anticoagulant and anti-inflammatory functions according to claim 1, characterized in that: The concentration of the polyphenol solution in step (1) is 1-4 mg / mL.

4. The method for preparing a super-hydrophilic coating having anticoagulant and anti-inflammatory functions according to claim 1, characterized in that: The substrate material in step (2) is a metal-based biomaterial, a polymer-based biomaterial or a decellularized biological tissue.

5. The method for preparing a super-hydrophilic coating having anticoagulant and anti-inflammatory functions according to claim 1, characterized in that: The magnesium-containing solution in step (3) is at least one of a magnesium sulfate solution, a magnesium chloride solution and a magnesium nitride solution.

6. The method for preparing a super-hydrophilic coating having anticoagulant and anti-inflammatory functions according to claim 1, characterized in that: The coating method in step (3) is drip coating, spray coating or dipping.

7. A super-hydrophilic coating with anticoagulant and anti-inflammatory functions, characterized in that: The method is prepared by the preparation method according to any one of claims 1 to 6.

8. Use of the super-hydrophilic coating with anticoagulant and anti-inflammatory functions as claimed in claim 7 in the preparation of cardiovascular materials / devices.

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