CN115607728B - Coating capable of releasing nitric oxide and resisting coagulation for long-term catalysis and preparation method thereof - Google Patents
Coating capable of releasing nitric oxide and resisting coagulation for long-term catalysis and preparation method thereof Download PDFInfo
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- CN115607728B CN115607728B CN202211266044.XA CN202211266044A CN115607728B CN 115607728 B CN115607728 B CN 115607728B CN 202211266044 A CN202211266044 A CN 202211266044A CN 115607728 B CN115607728 B CN 115607728B
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/02—Inorganic materials
- A61L27/04—Metals or alloys
- A61L27/06—Titanium or titanium alloys
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/28—Materials for coating prostheses
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/28—Materials for coating prostheses
- A61L27/34—Macromolecular materials
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/54—Biologically active materials, e.g. therapeutic substances
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- A61L33/00—Antithrombogenic treatment of surgical articles, e.g. sutures, catheters, prostheses, or of articles for the manipulation or conditioning of blood; Materials for such treatment
- A61L33/0005—Use of materials characterised by their function or physical properties
- A61L33/0011—Anticoagulant, e.g. heparin, platelet aggregation inhibitor, fibrinolytic agent, other than enzymes, attached to the substrate
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- A—HUMAN NECESSITIES
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L33/00—Antithrombogenic treatment of surgical articles, e.g. sutures, catheters, prostheses, or of articles for the manipulation or conditioning of blood; Materials for such treatment
- A61L33/0076—Chemical modification of the substrate
- A61L33/0088—Chemical modification of the substrate by grafting of a monomer onto the substrate
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L33/00—Antithrombogenic treatment of surgical articles, e.g. sutures, catheters, prostheses, or of articles for the manipulation or conditioning of blood; Materials for such treatment
- A61L33/06—Use of macromolecular materials
- A61L33/08—Polysaccharides
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- A—HUMAN NECESSITIES
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/216—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials with other specific functional groups, e.g. aldehydes, ketones, phenols, quaternary phosphonium groups
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- A—HUMAN NECESSITIES
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/23—Carbohydrates
- A61L2300/232—Monosaccharides, disaccharides, polysaccharides, lipopolysaccharides
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- A—HUMAN NECESSITIES
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/42—Anti-thrombotic agents, anticoagulants, anti-platelet agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2420/00—Materials or methods for coatings medical devices
- A61L2420/06—Coatings containing a mixture of two or more compounds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/40—Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
Abstract
The invention discloses a coating for releasing nitric oxide and resisting coagulation through long-term catalysis and a preparation method thereof, wherein the coating takes a metal-based biological material as a base layer and comprises anatase type TiO 2 The preparation method of the coating comprises the following steps of: taking a metal-based biological material as an anode, and adopting an electrochemical oxidation method to prepare regular TiO on the surface of the anode 2 And (3) drying and annealing the nanotube layer, performing hydrothermal treatment in a solution containing metal ions, and sequentially placing the nanotube layer in a mixed solution of a dopamine solution and lysine/heparin/selenocysteine for deposition reaction to obtain the coating. The coating prepared by the invention has excellent anticoagulation, biocompatibility and bioactivity, can realize long-term release of catalytic nitric oxide, promotes endothelialization of metal-based biological materials, ensures functional expression of normal endothelial cells, and has the advantages of simple, convenient and easy preparation method and good stability.
Description
Technical Field
The invention relates to a bio-based coating and a preparation method thereof, in particular to a coating capable of releasing nitric oxide and resisting coagulation for a long time in a catalytic manner and a preparation method thereof.
Background
Titanium alloy has excellent mechanical property, corrosion resistance and biocompatibility, so that the titanium alloy is applied to human tissue implantation substitute materials such as blood, orthopaedics, teeth and the like. Among blood contact materials, titanium and its alloys are commonly used for heart valves, vascular stents, blood circulation aids, and the like. However, the surfaces of titanium and titanium alloys are biologically inert, lack anticoagulant activity, and are incapable of inducing the formation of a natural anticoagulant-endothelial layer. When used as a long-term implantation material, a large amount of platelets and plasma proteins adhere to the surface of the implantation material, so that thrombus, vascular restenosis, inflammation and other reactions are caused, and the surface of the implantation material and human tissues have weak mutual binding capacity, so that implantation failure is caused. In addition, titanium alloys are required to be rapidly endothelialized as blood contact materials, and it is desirable that the surface of the titanium material promote functional expression of normal endothelial cells.
Healthy endothelial cells maintain vascular tone, inhibit platelet aggregation and activation, inhibit proliferation and migration of smooth muscle cells by synthesizing nitric oxide vasodilator. At present, nitric oxide signal molecules are introduced to the surface of biological materials, exogenous nitric oxide donor molecules can be fixed on the surface of the biological materials, and the signal molecules are slowly released after being implanted into the body. However, such a surface-mounted nitric oxide donor is very limited, and the purpose of releasing nitric oxide for a long period of time cannot be achieved. Human blood contains a large amount of endogenous nitric oxide carriers. The catalytic molecules for releasing nitric oxide molecules are fixed on the surface of the titanium material, so that the release of endogenous nitric oxide in human plasma can be catalyzed.
The invention patent with publication number CN102950102 discloses a preparation method of a titanium and titanium alloy surface multiple growth factor slow-release coating, which uses nano particles as a growth factor carrier, uses a charged polymer film as a medium, and prepares a multilayer coating containing nano particles of growth factors on the titanium surface. The nano particles contained in the coating are prepared by mixing polycation solution containing dopamine with a certain concentration and polyanion solution containing growth factors. The nano particles have good dispersibility, can protect the activity of growth factors, and are easy to fix on the surface of a material. By controlling the electrical properties of the nano-particles and the polymer film, the nano-particles carrying different factors are embedded between the polymer films. By adjusting the loading amount, loading sequence and coating thickness of different nano particles, the effective control of the release rate, release amount and release sequence of various growth factors can be realized. However, the preparation method of the patent is limited to bone tissue repair and bone material preparation, and the carried growth factors are also growth factors such as BMP2, TGF-beta, FGF, IGF and the like which can promote bones, and the preparation of cardiovascular materials is not mentioned.
The invention patent with publication number of CN108969805 discloses an anticoagulant hydrogel coating capable of catalyzing nitric oxide release and a preparation method thereof, and the patent comprises the following steps: (1) pre-treating a substrate material; (2) At room temperature, placing the pretreated substrate material in a mixed solution of poly-positive electrolyte, polyphenol and compound A in sequence, reacting for 1-20min in the poly-negative electrolyte, and then cleaning for 3-5 times; (3) And (3) at room temperature, taking the product obtained in the step (2) as a substrate, repeating the operation of the step (2) for 5-20 times, and drying by nitrogen to obtain the anticoagulation hydrogel coating capable of catalyzing nitric oxide release. The preparation of the patent adopts an electrostatic adsorption mode, and has a certain influence on the long-term stability of the coating. In addition, the electrostatically adsorbed catalyst is significantly limited in number.
Disclosure of Invention
The invention aims to: in order to solve the technical problems in the prior art, the invention aims to provide a coating which has excellent anticoagulation activity and biological activity and can realize long-term catalytic release of nitric oxide molecules, and also provides a preparation method of the coating.
The technical scheme is as follows: the long-term catalytic nitric oxide releasing and anticoagulation coating of the invention takes a metal-based biological material as a base layer and comprises anatase type TiO 2 Nanotube layer, metal ion layer, dopamine layer and lysine/heparin/selenocysteine combined layer, wherein the metal ion layer is carried on anatase type TiO 2 On the layer of the nano-tube,and then binds to the dopamine layer, which on the other side binds to the lysine/heparin/selenocysteine combination layer.
Further, the anatase type TiO 2 The inner diameter of the nano tube is 50-120nm, the tube length is 1-5 μm, and the metal ion is one or two of copper ion and zinc ion.
Further, the metal-based biomaterial is a titanium alloy, preferably one of a TA1 alloy, a TA2 alloy or a TC4 alloy.
The preparation method of the coating capable of releasing nitric oxide and resisting coagulation for a long time comprises the following steps:
(1) Taking a metal-based biological material as an anode, and adopting an electrochemical oxidation method to prepare regular TiO on the surface of the anode 2 The nanotube layer is dried and then annealed to obtain anatase type;
(2) Carrying out hydrothermal treatment on the material prepared in the step (1) in a solution containing metal ions;
(3) Placing the material prepared in the step (2) into a dopamine solution for deposition;
(4) Dissolving lysine in Tris buffer solution, adding heparin, fully stirring for reaction, then adding an activating agent for activation, and then adding selenocysteine for reaction to obtain a lysine/heparin/selenocysteine mixed solution;
(5) And (3) soaking the material prepared in the step (3) in the lysine/heparin/selenocysteine mixed solution prepared in the step (4), carrying out concussion reaction, washing with water, and drying to obtain the coating capable of catalyzing and releasing nitric oxide for a long time and resisting coagulation.
Further, in the step (1), the electrochemical oxidation conditions are as follows: in a constant voltage mode, the oxidation voltage is 30-100V, the oxidation time is 30-120min, the organic electrolyte is ethylene glycol+0.5wt% ammonium fluoride+2vol% deionized water, 0.5wt% HF or ethylene glycol+1.5wt% ammonium fluoride+5vol% deionized water, the annealing treatment temperature is 400-500 ℃, and the annealing treatment time is 1-5h.
Further, in the step (2), the concentration of the solution containing metal ions is 0.5-2.5mol/L, the temperature of the hydrothermal treatment is 150-200 ℃, and the time of the hydrothermal treatment is 1-5h.
Further, in the step (3), the pH of the dopamine solution is 8.0-8.5, and the soaking time is 10-24 hours.
Further, in the step (4), the mass ratio of lysine to heparin is 5:3-5:2, the concentration of lysine is 0.5-5mg/ml, preferably 2.5mg/ml, and the conditions for fully stirring and reacting the lysine and heparin are as follows: reacting for 1-5h in the environment of 2-8 ℃, wherein an activator is 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide and N-hydroxysuccinimide, the mol ratio of the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide to the N-hydroxysuccinimide is 3:1, the concentration of the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide is 1-3mg/ml, the preferential concentration is 2.5mg/ml, the activating time is 10-30min, the concentration of selenocysteine is 0.5-1.5mmol/L, and the shaking reaction time is 5-10h.
The principle of the invention: the invention prepares TiO by anodic oxidation annealing treatment 2 The inner diameter of the nanotube array is 50-120nm, and the length of the nanotube array is 1-5 mu m. Doping Cu in nano tube by hydrothermal method 2+ 、Zn 2+ One or more of the following. Studies have shown that both metal ions promote functional expression of normal endothelial cells, in which Cu 2+ Has obvious catalysis effect on the release of endogenous NO molecules in blood plasma. The nanotube array carrying the metal ions after being modified by the dopamine can play a role of an intermediate layer for fixing anticoagulant molecules and NO catalytic release molecules. Heparin has stronger anticoagulation, and is a commonly used medicine for anticoagulation and thrombosis inhibition in clinic. Selenocyamine is a small molecular compound containing selenium and plays an important role in catalyzing endogenous NO release. The characteristics of positive charge of lysine in solution and negative charge of heparin are utilized, the lysine and the heparin are assembled by static electricity, and then carboxyl of the lysine and amino of selenocysteine react with each other to generate amide so as to be combined covalently, so that the heparin and selenocysteine are assembled together; and then the surface of the titanium alloy is bonded through Schiff base reaction between a large amount of amino groups in lysine and selenocysteine and quinone groups of oxidized dopamine.
The beneficial effects are that: compared with the prior art, the invention has the following remarkable advantages:
(1) According to the invention, the regular nanotube array is prepared on the surface of the titanium alloy, so that the surface activity and the hydrophilicity of the titanium material are increased, and good sites are provided for subsequent ion and molecule loading; heparin molecules have good anticoagulation function, and carried metal ions and selenocysteine molecules have excellent capability of catalyzing endogenous NO donor release, so that the heparin molecule can realize various physiological functions as an implantation material and can be applied to blood contact materials;
(2) The invention can realize the simultaneous fixation of heparin and selenocysteine molecules on the surface of the titanium alloy through electrostatic self-assembly and covalent bonding;
(3) The preparation technology of the invention has mild required conditions, no special equipment, simple process and low cost.
Drawings
FIG. 1 is a drawing of a coated electron microscope prepared in example 2 of the present invention, wherein (a) and (c) are TiO anodized and annealed surfaces of a titanium alloy base layer 2 Nanotube layer, (b) and (d) are metal ion loaded active coatings with catalytic NO release;
fig. 2 is a water contact angle profile of the coating layer prepared in example 2 of the present invention, wherein (a) is a water contact angle formed by a titanium alloy base layer: 51.2 ° ± 0.07 °, (b) is a water contact angle with titanium alloy as a base layer, the coating being a metal ion loaded active coating having catalytic NO release: 13.2 ° ± 0.02 °;
FIG. 3 is a graph showing the number of platelet adhesion on the surface of the titanium alloy base layer and the coating layer prepared in example 2 of the present invention;
FIG. 4 shows VEGF expression after 1 day and 3 days of cell culture on the surface of the titanium alloy substrate and coating layer prepared in example 2 of the present invention;
FIG. 5 shows the NO release after 1 day and 3 days of cell culture on the surface of the titanium alloy substrate and coating layer prepared in example 2 of the present invention.
Detailed Description
The invention will be further described with reference to specific examples and figures.
Example 1: the long-term catalytic nitric oxide release and anticoagulation coatingA layer with TA1 alloy as base layer and anatase type TiO 2 A nanotube layer, a copper ion/zinc ion layer, a dopamine layer and a lysine/heparin/selenocysteine combination layer.
The preparation method of the coating comprises the following steps: TA1 alloy was prepared into 50 mm. Times.50 mm. Times.2.5 mm specimens, and the surfaces were polished by 320, 400, 600, 800, 1000# sandpaper, polished, HF+HNO 3 After electropolishing for 5s, washing with water and drying; oxidizing in ethylene glycol plus 0.5wt% ammonium fluoride plus 2vol.% deionized water electrolyte by adopting anodic oxidation, maintaining the voltage at 60V, oxidizing for 30min, respectively ultrasonically cleaning water, ethylene glycol and ethanol for 5min, and drying; subsequently, the sample is put into a reaction kettle, znCl is added 2 、CuCl 2 0.5M each, after reacting for 2 hours at 200 ℃, taking out and washing 3 times, and air-drying; a1 mg/ml solution of dopamine was prepared using Tris buffer and the pH of the solution was adjusted to 8.0. The prepared TiO loaded with metal ions 2 The nanotubes are placed in the dopamine solution to react for 12 hours at room temperature, and then washed and dried; adding 2.5 mg/mlL-lysine and 1.5mg/ml heparin sodium into Tris solution, stirring, placing in a 2-8 ℃ environment for 10 hours, taking out, adding 2.5mg/ml 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide and 12mg/ml N-hydroxysuccinimide, activating for 20 minutes, adding 1mM selenocyamine to prepare a mixed solution, placing the regular nanotube array carrying metal ions into the mixed solution, and fully reacting for 8 hours to obtain the nano-tube.
Example 2: the long-term nitric oxide releasing and anticoagulation coating of the invention takes TC4 alloy as a base layer and comprises anatase TiO 2 A nanotube layer, a copper ion layer, a dopamine layer and a lysine/heparin/selenocysteine combined layer.
The preparation method of the coating comprises the following steps: the TC4 alloy is prepared into a sample with the diameter of 10mm multiplied by 2.5mm, and the surface is polished by 320, 400, 600, 800 and 1000# sand paper and polished, and HF+HNO is added 3 After electropolishing for 3s, washing with water and drying; anodic oxidation is adopted, oxidation is carried out in 0.5wt% HF electrolyte, after the voltage is kept at 45V and oxidation is carried out for 1h, water, glycol and ethanol are respectively ultrasonically cleaned for 5min, and then dried; subsequently, the sample was placed in a reaction vessel, and 0.5M CuCl was added 2 At 2After reacting for 2 hours at 00 ℃, taking out, washing for 3 times, and air-drying; a1 mg/ml solution of dopamine was prepared using Tris buffer and the pH of the solution was adjusted to 8.5. Placing the prepared TiO2 nanotube carrying metal ions into the dopamine solution, reacting for 12 hours at room temperature, washing with water, and drying; 2.5 mg/mlL-lysine and 1.5mg/ml heparin sodium are added into Tris solution, stirred and placed in the environment of 2-8 ℃ for 10 hours, taken out, 2.5mg/ml 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide and 12mg/ml N-hydroxysuccinimide are added for 20 minutes, and 1mM selenocysteine is added for preparing a mixed solution. And then placing the regular nanotube array loaded with metal ions into the mixed solution, and fully reacting for 8 hours to obtain the nano-tube.
Example 3: the long-term catalytic nitric oxide releasing and anticoagulation coating of the invention takes TA2 medical alloy as a base layer and comprises anatase TiO 2 A nanotube layer, a copper ion/zinc ion layer, a dopamine layer and a lysine/heparin/selenocysteine combination layer.
The preparation method of the coating comprises the following steps: preparing TA2 medical titanium alloy into a sample with the thickness of 50mm multiplied by 2.5mm, polishing the surface by 320, 400, 600, 800 and 1000# abrasive paper, and polishing, and adding HF and HNO 3 After electropolishing for 5s, washing with water and drying; oxidizing in ethylene glycol+1.5wt% ammonium fluoride+5vol% deionized water electrolyte by adopting anodic oxidation, maintaining the voltage at 100V, oxidizing for 1h, respectively ultrasonically cleaning water, ethylene glycol and ethanol for 5min, and drying; subsequently, the sample is put into a reaction kettle, znCl is added 2 、CuCl 2 0.5M each, after 3 hours of reaction at 180 ℃, taking out and washing 3 times, and air-drying; a1 mg/ml solution of dopamine was prepared using Tris buffer and the pH of the solution was adjusted to 8.5. Placing the prepared TiO2 nanotube carrying metal ions into the dopamine solution, reacting for 12 hours at room temperature, washing with water, and drying; 2.5 mg/mlL-lysine and 1mg/ml heparin sodium are added into Tris solution, stirred and placed in an environment of 2-8 ℃ for 4 hours to be taken out, 2.5mg/ml 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide and 12mg/ml N-hydroxysuccinimide are added for 20 minutes to be activated, and 0.5mM selenocysteine is added to prepare a mixed solution. And then placing the regular nanotube array loaded with metal ions into the mixed solution, and fully reacting for 8 hours to obtain the nano-tube.
FIGS. 1 to 5 are data relating to the characteristics of the nanotube coating, platelet adhesion and endothelial cell function expression obtained in example 2. The nanotubes prepared using a voltage of 45V have a diameter of about 60-80nm and a tube length of about 3. Mu.m. After carrying metal ions, depositing dopamine and fixing lysine/heparin/selenocysteine combined layers, the diameter of the nanotube is obviously reduced. Because a large number of hydrophilic groups (such as-OH, -COOH and the like) are introduced on the surface of the titanium alloy, the water contact of the coating is obviously greatly reduced compared with the titanium alloy. The number of platelet adhesion on the coated surface is significantly reduced. After endothelial cells are inoculated on the surfaces of the titanium alloy and the coating and cultured for 1 and 3 days, the VEGF and NO concentration expressed by the cell function is obviously improved.
Claims (8)
1. A coating for long-term catalytic release of nitric oxide and anticoagulation, which uses metal-based biological material as base layer, is characterized in that the coating comprises anatase TiO 2 Nanotube layer, metal ion layer, dopamine layer and lysine/heparin/selenocysteine combined layer, wherein the metal ion layer is carried on anatase type TiO 2 The nanotube layer is then combined with the dopamine layer, and the other side of the dopamine layer is combined with the lysine/heparin/selenocysteine combined layer;
the preparation method of the coating for releasing nitric oxide and resisting coagulation by long-term catalysis comprises the following steps:
(1) Taking a metal-based biological material as an anode, and adopting an electrochemical oxidation method to prepare regular TiO on the surface of the anode 2 The nanotube layer is dried and then annealed to obtain anatase type;
(2) Carrying out hydrothermal treatment on the material prepared in the step (1) in a solution containing metal ions;
(3) Placing the material prepared in the step (2) into a dopamine solution for deposition reaction;
(4) Dissolving lysine in Tris buffer solution, adding heparin, fully stirring for reaction, then adding an activating agent for activation, and then adding selenocysteine for reaction to obtain a lysine/heparin/selenocysteine mixed solution;
(5) Placing the material prepared in the step (3) into the lysine/heparin/selenocysteine mixed solution prepared in the step (4), carrying out concussion reaction, washing with water, and drying to obtain a coating capable of catalyzing and releasing nitric oxide for a long time and resisting coagulation;
the metal ions are one or two of copper ions and zinc ions.
2. The coating of claim 1, wherein the anatase TiO 2 The inner diameter of the nano tube is 50-120nm, and the tube length is 1-5 μm.
3. The coating of claim 1, wherein in step (1), the electrochemical oxidation conditions are: the oxidation voltage is 30-100V, the oxidation time is 30-120min, the annealing treatment temperature is 400-500 ℃, and the annealing treatment time is 1-5h.
4. The coating of claim 1, wherein in step (2), the concentration of the metal ion-containing solution is 0.5-2.5mol/L, the temperature of the hydrothermal treatment is 150-200 ℃, and the time of the hydrothermal treatment is 1-5h.
5. The coating of claim 1, wherein in step (4), the mass ratio of lysine to heparin is 5:3-5:2, and the concentration of lysine is 0.5-5mg/ml.
6. The coating of claim 1, wherein in step (4), the reaction conditions of lysine with heparin are: and (3) in the environment of 2-8 ℃ for 1-5h.
7. The coating of claim 1, wherein in step (4), the activator is 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide and N-hydroxysuccinimide, and the activation time is 10-30min.
8. The coating of claim 1, wherein in step (4), the concentration of selenocysteine is 0.5-1.5mmol/L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211266044.XA CN115607728B (en) | 2022-10-17 | 2022-10-17 | Coating capable of releasing nitric oxide and resisting coagulation for long-term catalysis and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211266044.XA CN115607728B (en) | 2022-10-17 | 2022-10-17 | Coating capable of releasing nitric oxide and resisting coagulation for long-term catalysis and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115607728A CN115607728A (en) | 2023-01-17 |
| CN115607728B true CN115607728B (en) | 2023-07-21 |
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