CN114588314A

CN114588314A - Vascular stent and preparation method and application thereof

Info

Publication number: CN114588314A
Application number: CN202210337282.9A
Authority: CN
Inventors: 杨志禄; 王文轩; 牟小辉; 杜泽煜; 王颖; 张文泰
Original assignee: Dongguan Peoples Hospital
Current assignee: Dongguan Peoples Hospital
Priority date: 2022-03-31
Filing date: 2022-03-31
Publication date: 2022-06-07
Anticipated expiration: 2042-03-31
Also published as: CN114588314B

Abstract

The invention discloses a vascular stent and a preparation method and application thereof, and relates to the technical field of preparation of biomedical materials. The blood vessel support is formed by sequentially depositing nitric oxide on the surface of the blood vessel support to catalyze and release micro-nano particles and thrombin inhibitor, and performing surface modification on the blood vessel support. The nitrogen monoxide catalytic release micro-nano particles have stable chemical structure at room temperature and normal pressure, the nitrogen monoxide catalytically released by the nitrogen monoxide catalytic release micro-nano particles can inhibit the activation and adhesion of platelets, and meanwhile, active reaction functional groups on the surfaces of the nitrogen monoxide catalytic release micro-nano particles can be grafted or grafted, so that the fixation of the nitrogen monoxide catalytic release micro-nano particles on the surface of a stent and the effective load and controllable elution of a medicine on the surfaces of the micro-nano particles are realized, and the slow release effect is improved. In addition, the thrombin inhibitor is fixed in an adsorption mode, so that the elution time of the thrombin inhibitor can be prolonged, and the controllable elution of the thrombin inhibitor is realized.

Description

Intravascular stent and preparation method and application thereof

Technical Field

The invention relates to the technical field of biomedical material preparation, in particular to a vascular stent and a preparation method and application thereof.

Background

At present, metal-based vascular stents have been developed as the primary means of treatment for cardiovascular disease. The expansion of the stent can cause damage to vascular tissues at the focus part, so that thrombus is formed, the health of a patient is seriously harmed, and even the life safety is endangered. Currently, there are two main strategies to solve this problem, drug therapy and stent surface modification.

The drug therapy mainly comprises injection of anticoagulant, oral administration of anticoagulant, use of platelet antagonist, use of thrombolytic drugs and the like, but the use of drugs is accompanied by risks of side effects such as bleeding, drug resistance and gastrointestinal reactions.

The surface modification of the stent mainly comprises an inorganic film coating, a drug blending organic coating, an electrostatic self-assembly fixed anticoagulant molecule and a chemical coupling fixed anticoagulant molecule. The inorganic film coating has good anticoagulation function, but poor flexibility, and is easy to fall off in the processes of stent crimping, pushing and expanding to cause thrombus to cause harm; the drug blending organic coating can effectively keep the activity of anticoagulant molecules, but the release rate is too fast, so that the anticoagulant coating is rapidly ineffective; the chemical coupling and fixing of the anticoagulant molecules can effectively improve the utilization rate and chemical stability of the anticoagulant molecules, but can influence the activity of the anticoagulant molecules to a certain degree; the electrostatic self-assembly fixed anticoagulant molecules can also realize the surface fixation of the anticoagulant molecules, can retain the activity of the anticoagulant molecules to a greater extent, but have poorer chemical stability and environmental stability than the anticoagulant molecules fixed by a chemical coupling strategy.

In view of this, the invention is particularly proposed.

Disclosure of Invention

The invention aims to provide a preparation method of a vascular stent, the vascular stent and application of the vascular stent in the field of drug loading.

The invention is realized by the following steps:

in a first aspect, the invention provides a preparation method of a vascular stent, which comprises the steps of depositing nitric oxide on the surface of the vascular stent in sequence to catalyze and release micro-nano particles and thrombin inhibitor, and carrying out surface modification on the vascular stent. The nitric oxide catalytic release micro-nano particles comprise a compound framework and a compound doped in the compound framework and having a nitric oxide catalytic release function, and the compound framework comprises one or more of phenolic compounds, quinone compounds and ketone compounds. The immobilization mode for the deposition of thrombin inhibitors is adsorption.

In a second aspect, the present invention provides a vascular stent made by the method of any one of the preceding embodiments.

In a third aspect, the present invention provides the use of a vascular stent as in the previous embodiments in the manufacture of a vascular repair article or an animal model of vascular repair.

The invention has the following beneficial effects:

the invention provides a vascular stent with nitric oxide catalytic release and thrombin inhibitor controllable elution, and a preparation method and application thereof. Meanwhile, the compound doped in the material and having the nitric oxide catalytic release function can catalyze the release of nitric oxide, so that the formation of thrombus is further inhibited. In addition, the thrombin inhibitor is fixed on the surface of the blood vessel stent in an adsorption mode, and compared with a covalent grafting method, the thrombin inhibitor can be adsorbed and fixed, so that the elution time of the thrombin inhibitor can be prolonged, and the controllable elution of the thrombin inhibitor is realized.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.

FIG. 1 is a transmission electron microscope image of the nitric oxide catalyzed micro-nano particle prepared in example 1;

FIG. 2 is a transmission electron microscope energy spectrum diagram of the nitrogen monoxide catalyzed micro-nano particle prepared in example 1;

FIG. 3 is a transmission electron microscope energy spectrum diagram of the nitrogen monoxide catalyzed micro-nano particle prepared in example 1;

FIG. 4 is a transmission electron microscope energy spectrum diagram of the nitrogen monoxide catalyzed micro-nano particle prepared in example 1 for releasing Se atoms;

FIG. 5 is a transmission electron microscope energy spectrum diagram of nitrogen atoms in the nitrogen monoxide catalyzed micro-nano particles prepared in example 1;

FIG. 6 is a nitric oxide catalyzed release rate profile of a vascular stent prepared in example 1;

FIG. 7 is a graph showing the loading rate of the NO catalytic release micro-nanoparticle carrier compound prepared in example 1;

FIG. 8 is a graph showing the nitric oxide release rate of the vascular stent prepared in each step of example 1;

FIG. 9 is a graph showing the release rate of the stent prepared in example 1;

FIG. 10 is a blood coagulation time chart of the blood vessel stent prepared in each step of example 1;

FIG. 11 is a diagram of a centrifuged sample of the nitric oxide-catalyzed micro/nano particle deposition solution prepared in example 1;

FIG. 12 is a diagram of a centrifuged sample of the nitric oxide-catalyzed micro/nano particle deposition solution prepared in comparative example 1;

FIG. 13 is a transmission electron microscope energy spectrum of blank 316L stainless steel sheet cultured endothelial cells;

FIG. 14 is a transmission electron microscopy energy spectrum of endothelial cells cultured on 316L stainless steel sheets prepared by the method of example 1;

FIG. 15 is a transmission electron microscope energy spectrum of endothelial cells cultured on 316L stainless steel sheets prepared by the method of comparative example 2;

FIG. 16 is a histogram of the number of endothelial cells cultured in test example 2.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.

In some vascular repair products or cardiovascular diseases, the use of vascular stents has become a main means, but vascular stents still have vascular tissue damage at a focus part caused by stent expansion in the actual use process, further cause thrombosis, and seriously harm the health and life safety of patients. In order to solve the above problems, the inventors propose to sequentially deposit a nanoparticle having a nitric oxide catalytic release function and a thrombin inhibitor on the surface of a vascular stent to reduce thrombus formation.

Nitric Oxide (NO) is a key factor in maintaining the balance of the cardiovascular system as a signaling molecule, and has the effect of inhibiting activation and aggregation of platelets. The NO anticoagulant material mainly comprises an NO release type and an NO catalytic type, wherein the NO release type is limited in research and application due to short half-life period, high release speed and limited load capacity of an NO donor; the NO catalytic anticoagulant material can realize effective release within a controllable speed range by utilizing the function of decomposing endogenous nitric oxide donors with relatively stable abundance in vivo, and avoids the problems of burst release and limited load capacity of the NO release anticoagulant material.

The thrombin inhibitor is a direct thrombin inhibitor, has high specificity, short half-life period and flexible administration, can effectively reduce bleeding rate and metabolic burden, and can improve the utilization rate of the thrombin inhibitor and reduce metabolic pressure through a drug loading system.

According to the invention, one or more of phenolic compounds, quinone compounds and ketone compounds are used as compound frameworks, the nitric oxide catalytic release micro-nano particles obtained by doping the compound with the nitric oxide catalytic release function have stable chemical structure at room temperature and normal pressure, active reaction functional groups on the surface can be grafted with other molecules or grafted to the surfaces of other materials, so that thrombin inhibitors can be effectively loaded and controlled to be released, controllable elution of medicines is realized, the release time of the medicines is increased, and the slow release effect is improved. Meanwhile, the molecules or ions with nitric oxide catalytic release function doped inside can catalytically release nitric oxide, so that the formation of thrombus is further inhibited. In addition, the thrombin inhibitor is fixed on the surface of the blood vessel stent in an adsorption mode, and compared with a covalent grafting method, the thrombin inhibitor can be adsorbed and fixed, so that the elution time of the thrombin inhibitor can be prolonged, and the controllable elution of the thrombin inhibitor is realized.

In an optional embodiment, the nitric oxide catalytic release micro-nano particles are obtained by mixing the solution A, the solution B and the solution C, reacting, and performing solid-liquid separation after the reaction is finished.

The solution A is prepared by dissolving one or more of phenolic compounds, quinone compounds and ketone compounds in a dispersion medium; the solution B is prepared by dispersing a compound with the nitric oxide catalytic release function in a dispersion medium; the solution C is prepared by dissolving a compound containing copper ions in a dispersion medium.

The solution A provides a compound skeleton for a compound with a nitric oxide catalytic release function, the solution B and the solution C can catalytically release NO, wherein the solution B and the solution A can form micro-nano particles after reaction, and the solution C is mixed in the micro-nano particles formed by the solution B and the solution A in a doping mode to improve the release rate of nitric oxide.

Preferably, the solution a, the solution B and the solution C are mixed in a ratio of 1: 0.001 to 100: 0 to 100. More preferably, the solution a, the solution B and the solution C are mixed in a ratio of 1: 0.001-10: 0 to 1.

It can be understood that, since the micro-nano particles obtained after the reaction of the solution B and the solution a can also catalyze the release of nitric oxide, when the release rate of the micro-nano particles obtained after the reaction of the solution B and the solution a is high, the addition amount of the solution C may be 0. Similarly, when the release rate of the micro-nano particle nitric oxide obtained by doping the solution C in the solution a is high, the addition amount of the solution B can be 0.

Preferably, the concentration of the solution A, the concentration of the solution B and the concentration of the solution C are all 0.001-100 mg/mL.

More preferably, the concentration of the solution A is 0.001-10 mg/mL, the concentration of the solution B is 0.001-1 mg/mL, and the concentration of the solution C is 0.001-0.1 mg/mL.

Preferably, the reaction is a stirring or oscillation reaction, the reaction temperature is 0-80 ℃, and the reaction time is 0.01-72 hours. More preferably, the reaction temperature is 4-40 ℃ and the reaction time is 2-24 h.

The reason why the mixing reaction time can be 0.01h is that bonding reaction and non-bonding reaction can occur after the solution A, the solution B and the solution C are mixed, turbidity can be generated as long as the three solutions are mixed, the formation speed of the nitrogen monoxide catalytic release micro-nano particles is extremely high, the non-bonding reaction can occur in a very short time, and the micro-nano particles obtained by separation are more in non-chemical bonding and less in chemical bonding, but the nitrogen monoxide catalytic release micro-nano particles are also formed. The reaction time is prolonged, the probability of bonding reaction is increased, and the chemical bonding of the micro-nano particles obtained by re-separation is increased.

In an alternative embodiment, the phenolic compound comprises at least one of monophenol and derivatives thereof, catechol and derivatives thereof, pyrogallol and derivatives thereof, and phloroglucinol and derivatives thereof.

Preferably, the quinone compound includes at least one of benzoquinone and its derivatives, naphthoquinone and its derivatives, phenanthrenequinone and its derivatives, and anthraquinone and its derivatives.

Preferably, the ketone compound includes at least one of a flavonoid compound, a flavonol compound and a dihydroflavonoid compound.

Preferably, the compound having a catalytic nitric oxide releasing function comprises ions and/or molecules having a catalytic nitric oxide releasing function.

Preferably, the ions having nitric oxide catalytic release function include copper ion compounds or organic copper ions; more preferably, the copper ion is Cu⁺Or Cu²⁺。

Preferably, the molecule with nitric oxide catalytic release function comprises one or more of a compound containing thiol or disulfide bond or sulfur selenium bond or sulfur tellurium bond, selenol or diselenide bond or selenium tellurium compound, and tellurol or ditelluride bond, of which both ends are primary amino groups.

The phenolic compound, the quinone compound, the ketone compound and the compound with the nitric oxide catalytic release function are selected, the raw material sources are wide, the price is low, the cost of the surface deposition of the vascular stent is reduced, and the anticoagulation capability of the vascular stent is improved.

In an alternative embodiment, the method for fixing nitric oxide on the surface of a vascular stent to catalytically release micro-nano particles comprises the following steps: dispersing the nitric oxide catalytic release micro-nano particles in a deposition medium to obtain nitric oxide catalytic release micro-nano particle deposition solution, and then putting the intravascular stent into the nitric oxide catalytic release micro-nano particle deposition solution for deposition to obtain the intravascular stent with the nitric oxide catalytic release function.

Preferably, the immobilization mode of the nitric oxide catalytic release micro-nano particle deposition comprises any one of adsorption, electrostatic interaction, covalent grafting or bridging anchoring.

Preferably, the concentration of the nitric oxide catalyzed release micro-nano particle deposition solution is 0.001-100 mg/mL, and more preferably 0.05-10 mg/mL.

The fixing mode of the nitric oxide catalytic release micro-nano particles on the vascular stent, such as adsorption, electrostatic action, covalent grafting or bridging anchoring, can be changed by controlling the concentration of the nitric oxide catalytic release micro-nano particle deposition solution, so that the bonding strength, density and the like of the nitric oxide catalytic release micro-nano particles fixed on the surface of the vascular stent are changed, and the catalytic release of nitric oxide in the vascular stent made of different materials is realized.

The adsorption is realized by non-bonding forces such as coordination, hydrogen bonds, intermolecular force, hydrophobic force and the like, the micro-nano particles are fixed on the surface to be modified, the generation speed of the micro-nano particles is higher than that of the bonding, for example, on the surface of a 316L SS intravascular stent, phenolic hydroxyl on the surface of the micro-nano particles catalytically released by nitric oxide can generate coordination with metal atoms on the surface of the 316L SS intravascular stent, the process is higher than that of the bonding, and the nitrogen monoxide catalytic release function is endowed to the 316L SS intravascular stent after the micro-nano particles are fixed.

Covalent grafting is to perform bonding chemical reaction on phenolic hydroxyl, ketone group, aldehyde group and aromatic ring on the surface of the micro-nano particle, for example, on the surface of an aminated blood vessel stent, the stent can perform Schiff base reaction with the phenolic hydroxyl on the surface of the micro-nano particle released by nitric oxide catalysis or perform Michael addition reaction on a benzene ring where the phenolic hydroxyl is positioned, so as to complete covalent grafting, the generation speed of the process is slower than that of a non-bonding action process, the grafted blood vessel stent has better chemical stability and action environment stability compared with non-bonding action fixation, and the aminated blood vessel stent is endowed with the nitric oxide catalysis release function after fixation.

Preferably, the deposition medium comprises one or more of distilled water with pH of 4-12, PBS buffer, Tris-hydrochloric acid buffer, HEPES buffer, ethanol aqueous solution, tetrahydrofuran aqueous solution, acetone, benzene and its derivatives, dimethyl sulfoxide, dimethyl formamide and acetonitrile.

The deposition temperature is 4-80 ℃, and the deposition time is 0.01-72 h; preferably, the deposition temperature is 4-40 ℃, and the deposition time is 0.5-24 h.

The nitrogen monoxide catalytic release micro-nano particles prepared by the method have stable chemical structure at room temperature and normal pressure, and active reaction functional groups on the surface can be grafted with other molecules and other material surfaces; the compound with the nitric oxide catalytic release function doped on the surface and in the micro-nano particles is endowed with the function of catalytically releasing nitric oxide by the nitric oxide catalytic release micro-nano particles, and the catalytic release rate of the compound can be regulated and controlled by the doping amount of nitric oxide.

In an alternative embodiment, a method of depositing a thrombin inhibitor on the surface of a vascular stent comprises: and (3) placing the intravascular stent fixed with the nitric oxide catalytic release micro-nano particles in a thrombin inhibitor solution for deposition to obtain the intravascular stent with the nitric oxide catalytic release function and the controllable elution of the thrombin inhibitor.

The thrombin inhibitor solution is prepared by dispersing a thrombin inhibitor into a dispersion medium to prepare the thrombin inhibitor solution with the concentration of 0.001-100 mg/mL.

Preferably, the concentration of the thrombin inhibitor solution is 0.001-10 mg/mL, more preferably 0.05-5 mg/mL. By controlling the concentration of the thrombin inhibitor within the range, the adsorption and fixation of the thrombin inhibitor on the surface of the blood vessel stent on which the nitric oxide catalytic release micro-nano particles are fixed can be realized, so that the elution process of the thrombin inhibitor can be sustainable, and the controllable elution of the thrombin inhibitor on the surface of the blood vessel stent is realized.

Preferably, the deposition temperature is 4-80 ℃, the deposition time is 0.01-72 h, more preferably, the deposition temperature is 4-37 ℃, and the deposition time is 0.5-24 h.

It should be noted that the deposition process of the thrombin inhibitor and the nitric oxide catalytic release micro-nano particles is mainly a non-bonding reaction, and the non-bonding reaction can occur within a very short time range, so that when the vascular stent is contacted with the nitric oxide catalytic release micro-nano particle deposition solution or the thrombin inhibitor solution, the surface modification process of the nitric oxide catalytic release micro-nano particles or the thrombin inhibitor is already formed, the deposition time of the two can approach zero, and the loading amounts of the nitric oxide catalytic release micro-nano particles and the thrombin inhibitor can be controlled by controlling the length of the reaction time.

In alternative embodiments, the thrombin inhibitor comprises a direct thrombin inhibitor or an indirect thrombin inhibitor.

Preferably, the direct thrombin inhibitor comprises one or more of dabigatran etexilate, bivalirudin, argatroban and recombinant hirudin.

Preferably, the indirect thrombin inhibitor comprises one or more of heparin, low molecular weight heparin, enoxaparin and nadroparin.

In an alternative embodiment, to facilitate the preparation of the vascular stent, the dispersion medium may be a conventional medium, the buffer solution comprises one or more of distilled water with the pH value of 2-14, a barbital sodium buffer solution, a barbital sodium-hydrochloric acid buffer solution, a glycine-sodium hydroxide buffer solution, a glycine-hydrochloric acid buffer solution, a HEPES buffer solution, a phthalic acid-hydrochloric acid buffer solution, a potassium dihydrogen phosphate-sodium hydroxide buffer solution, a phosphate buffer solution, a citric acid-phosphoric acid buffer solution, a citric acid-sodium citrate buffer solution, a citric acid-sodium hydroxide-hydrochloric acid buffer solution, a PBS buffer solution, a borax-boric acid buffer solution, a borax-sodium hydroxide buffer solution, a TAE buffer solution, a sodium carbonate-sodium bicarbonate buffer solution, a TBST buffer solution, a TE buffer solution, a TEN buffer solution, a Tris-hydrochloric acid buffer solution and an acetic acid-sodium acetate buffer solution.

In alternative embodiments, the material of the vascular stent is metal or polymer according to different application requirements.

Preferably, the metal comprises one or more of 316L SS, Ti and alloys thereof, Ti-O, Ti-N, Ti-Ni, Co-Cr alloys, Fe and alloys thereof, Zn and alloys thereof, and Mg and alloys thereof;

preferably, the polymer comprises one or more of a homopolymer or copolymer of polylactic acid, polyglycolic acid, polycaprolactone, and polyanhydride.

The nitric oxide on the surface of the intravascular stent provided by the invention catalyzes and releases active reaction functional groups on the surface of the micro-nano particles, so that other molecules can be grafted and grafted to the surfaces of other materials, and drugs can be effectively loaded and released under control. Therefore, in a specific application process, the carrier can be used as a drug loading carrier in the fields of vascular repair products or vascular repair animal models and the like, and the loadable drugs comprise anticoagulant drugs, anticancer drugs, antibacterial drugs, anti-inflammatory drugs, anti-immunogenicity drugs, osteoinductive drugs and the like.

The features and properties of the present invention are described in further detail below with reference to examples.

Example 1

The embodiment provides a vascular stent, which comprises steps of adsorbing selenium-containing nitric oxide catalytic release micro-nano particles synthesized by tannic acid and selenocysteine on the surface of a 316L SS vascular stent, and then depositing a bivalirudin thrombin inhibitor, wherein the steps of the preparation are as follows:

s1 preparation of nitrogen monoxide catalytic release micro-nano particles

Tannic acid was weighed and dissolved in PBS buffer solution of pH 8 to prepare tannic acid solution A of 1 mg/mL.

Selenium-containing selenocysteine molecules are weighed and dispersed in PBS buffer solution with the pH value of 8 to prepare the selenocysteine solution B with the concentration of 0.1 mg/mL.

Copper chloride containing copper ions was weighed and dispersed in PBS buffer solution of pH 8 to prepare copper chloride solution C with a concentration of 0.1 mg/mL.

And (3) mixing the solution A, the solution B and the solution C according to the mass ratio of 1: 1: 0 mixing, stirring at 40 ℃ or reacting for 24 hours by oscillation; after the reaction is finished, separating to obtain the micro-nano particles which take polyphenol tannic acid as a structural basis and are catalyzed and released by nitric oxide doped with selenium-containing selenocysteamine molecules.

S2 deposition of micro-nano particles released by nitric oxide catalysis

Weighing the nitric oxide catalytic release micro-nano particles obtained in the step S1, dispersing the nitric oxide catalytic release micro-nano particles into PBS buffer solution to prepare nitric oxide catalytic release micro-nano particle deposition solution with the concentration of 10mg/mL, soaking the vascular stent into the nitric oxide catalytic release micro-nano particle deposition solution, reacting for 24 hours at 40 ℃, and after the reaction is finished, fully rinsing and drying the vascular stent by using distilled water to obtain the vascular stent with the surface deposited with the nitric oxide catalytic release micro-nano particles.

S3, deposition of Thrombin inhibitors

Weighing bivalirudin, dispersing the bivalirudin in distilled water to prepare a bivalirudin solution with the concentration of 1mg/mL, soaking the vascular stent which is obtained in the step S2 and has the surface deposited with the nitric oxide to catalyze and release the micro-nano particles in the bivalirudin solution, depositing the vascular stent at 37 ℃ for 6 hours, and after the reaction is finished, fully rinsing the vascular stent with distilled water and drying the vascular stent to obtain the bivalirudin preparation.

Example 2

The embodiment provides a blood vessel stent, which comprises the steps of adsorbing copper ion-doped nitric oxide catalytic release micro-nano particles synthesized by tannic acid, selenocysteine and copper chloride on the surface of a 316L SS blood vessel stent, and then depositing a bivalirudin thrombin inhibitor, wherein the preparation steps are as follows:

s1 preparation of nitrogen monoxide catalytic release micro-nano particles

Copper chloride was weighed and dispersed in PBS buffer solution at pH 8 to prepare copper chloride solution C at a concentration of 0.01 mg/mL.

And (3) mixing the solution A, the solution B and the solution C according to the mass ratio of 1: 0.01: 0.1, mixing, stirring at 40 ℃ or carrying out oscillation reaction for 24 hours; after the reaction is finished, separating to obtain the selenium-containing selenocysteamine molecules which take polyphenol tannic acid as a structural basis and catalyze and release the micro-nano particles by nitric oxide doped with copper ions.

S2 deposition of micro-nano particles released by nitric oxide catalysis

Weighing the nitric oxide catalytic release micro-nano particles obtained in the step S1, dispersing the nitric oxide catalytic release micro-nano particles into PBS buffer solution to prepare nitric oxide catalytic release micro-nano particle deposition solution with the concentration of 0.1mg/mL, soaking the vascular stent into the nitric oxide catalytic release micro-nano particle deposition solution, reacting for 24 hours at 25 ℃, and after the reaction is finished, fully rinsing and drying the vascular stent by using distilled water to obtain the vascular stent with the surface deposited with the nitric oxide catalytic release micro-nano particles.

S3, deposition of Thrombin inhibitors

Weighing bivalirudin, dispersing the bivalirudin in distilled water to prepare a bivalirudin solution with the concentration of 0.05mg/mL, soaking the blood vessel stent obtained in the step S2 in the bivalirudin solution, depositing for 12 hours at 25 ℃, after the reaction is finished, fully rinsing with distilled water, and drying to obtain the blood vessel stent.

Example 3

The embodiment provides a vascular stent, which comprises the steps of adsorbing nitrogen monoxide synthesized by tannic acid, copper chloride and selenocysteine on the surface of a Co-Cr alloy vascular stent for catalytic release of micro-nano particles, and then depositing a bivalirudin thrombin inhibitor, wherein the preparation steps are as follows:

s1 preparation of nitrogen monoxide catalytic release micro-nano particles

And (3) mixing the solution A, the solution B and the solution C according to the mass ratio of 1: 1: 0.1, mixing, stirring at 25 ℃ or carrying out oscillation reaction for 48 hours; after the reaction is finished, separating to obtain the micro-nano particles which take polyphenol tannic acid as a structural basis and contain selenium-containing selenocysteamine molecules and nitric oxide doped with copper ions for catalytic release.

S2 deposition of micro-nano particles released by nitric oxide catalysis

Weighing the nitric oxide catalytic release micro-nano particles obtained in the step S1, dispersing the nitric oxide catalytic release micro-nano particles into a Tris-hydrochloric acid buffer solution to prepare a nitric oxide catalytic release micro-nano particle deposition solution with the concentration of 10mg/mL, soaking the vascular stent into the nitric oxide catalytic release micro-nano particle deposition solution, reacting for 24 hours at 4 ℃, and after the reaction is finished, fully rinsing and drying the vascular stent by using distilled water to obtain the vascular stent with the surface deposited with the nitric oxide catalytic release micro-nano particles.

S3, deposition of Thrombin inhibitors

Weighing bivalirudin, dispersing the bivalirudin in distilled water to prepare a bivalirudin solution with the concentration of 1mg/mL, soaking the blood vessel stent obtained in the step S2 in the bivalirudin solution, depositing the blood vessel stent at 4 ℃ for 24 hours, fully rinsing the blood vessel stent with distilled water after the reaction is finished, and drying the blood vessel stent to obtain the blood vessel stent.

Example 4

The embodiment provides a vascular stent, which comprises the steps of adsorbing nitric oxide synthesized by tannic acid and cystamine on the surface of a 316L SS vascular stent to catalytically release micro-nano particles, and then depositing a bivalirudin thrombin inhibitor, wherein the preparation steps are as follows:

s1 preparation of nitrogen monoxide catalytic release micro-nano particles

Tannic acid was weighed and dissolved in HEPES buffer solution having a pH of 8 to prepare a tannic acid solution A having a concentration of 1 mg/mL.

Weighing sulfur-containing cystamine molecules, and dispersing the sulfur-containing cystamine molecules in HEPES buffer solution with the pH value of 8 to prepare cystamine solution B with the concentration of 0.1 mg/mL.

Copper chloride was weighed and dispersed in HEPES buffer solution at pH 8 to prepare copper chloride solution C at a concentration of 0.01 mg/mL.

And (3) mixing the solution A, the solution B and the solution C according to the mass ratio of 1: 1: 0, mixing, stirring at 40 ℃ or carrying out oscillation reaction for 2 hours; after the reaction is finished, separating to obtain micro-nano particles which take polyphenol tannic acid as a structural basis and catalytically release nitrogen monoxide doped with sulfur-containing cystamine molecules.

S2 deposition of micro-nano particles released by nitric oxide catalysis

Weighing the nitric oxide catalytic release micro-nano particles obtained in the step S1, dispersing the nitric oxide catalytic release micro-nano particles into PBS buffer solution to prepare nitric oxide catalytic release micro-nano particle deposition solution with the concentration of 10mg/mL, soaking the vascular stent into the nitric oxide catalytic release micro-nano particle deposition solution, reacting for 24 hours at 4 ℃, and after the reaction is finished, fully rinsing and drying the vascular stent by using distilled water to obtain the vascular stent with the surface deposited with the nitric oxide catalytic release micro-nano particles.

S3, deposition of Thrombin inhibitors

Weighing bivalirudin, dispersing the bivalirudin in distilled water to prepare a bivalirudin solution with the concentration of 1mg/mL, soaking the blood vessel stent obtained in the step S2 in the bivalirudin solution, depositing for 12 hours at 25 ℃, after the reaction is finished, fully rinsing with distilled water, and drying to obtain the blood vessel stent.

Example 5

The embodiment provides a vascular stent, which comprises the steps of adsorbing micro-nano particles synthesized by catechol derivatives caffeic acid and cysteamine telluride on the surface of a 316L SS vascular stent for catalyzing and releasing nitric oxide, and then depositing a bivalirudin thrombin inhibitor, wherein the preparation steps are as follows:

s1 preparation of nitrogen monoxide catalytic release micro-nano particles

Caffeic acid was weighed and dissolved in PBS buffer solution with pH 8 to prepare caffeic acid solution A with concentration of 1 mg/mL.

Weighing tellurium-containing cysteamine molecules, and dispersing the tellurium-containing cysteamine molecules in PBS buffer solution with the pH value of 8 to prepare 0.1mg/mL tellurium-containing cysteamine solution B.

And (3) mixing the solution A, the solution B and the solution C according to the mass ratio of 1: 0.1: 0, mixing, stirring at 15 ℃ or carrying out oscillation reaction for 16 hours; after the reaction is finished, separating to obtain the tellurium-containing micro/nano particles which are based on the structure of catechol caffeic acid and catalytically released by nitric oxide doped with tellurium-containing cysteamine telluride molecules.

S2 deposition of micro-nano particles released by nitric oxide catalysis

Weighing the nitric oxide catalytic release micro-nano particles obtained in the step S1, dispersing the nitric oxide catalytic release micro-nano particles into a sodium carbonate-sodium bicarbonate buffer solution to prepare a nitric oxide catalytic release micro-nano particle deposition solution with the concentration of 10mg/mL, soaking the vascular stent into the nitric oxide catalytic release micro-nano particle deposition solution, reacting for 12 hours at 4 ℃, and after the reaction is finished, fully rinsing and drying the vascular stent by using distilled water to obtain the vascular stent with the surface deposited with the nitric oxide catalytic release micro-nano particles.

S3, deposition of Thrombin inhibitors

Weighing bivalirudin, dispersing the bivalirudin in distilled water to prepare a bivalirudin solution with the concentration of 1mg/mL, soaking the blood vessel stent obtained in the step S2 in the bivalirudin solution, depositing for 24 hours at 4 ℃, fully rinsing with distilled water after the reaction is finished, and drying to obtain the blood vessel stent.

Example 6

The embodiment provides a vascular stent, which comprises the steps of adsorbing nitric oxide catalyzed release micro-nano particles synthesized by catechol derivatives dopamine and selenocysteine on the surface of a 316L SS vascular stent, and then depositing a bivalirudin thrombin inhibitor, wherein the preparation steps are as follows:

s1 preparation of nitrogen monoxide catalytic release micro-nano particles

Weighing dopamine, dissolving the dopamine into Tris-hydrochloride buffer solution with the pH value of 8, and preparing dopamine solution A with the concentration of 1 mg/mL.

And (3) mixing the solution A, the solution B and the solution C according to the mass ratio of 1: 1: 0, mixing, stirring at 40 ℃ or carrying out oscillation reaction for 24 hours; after the reaction is finished, separating to obtain the micro-nano particles which take catechol derivative dopamine as a structural basis and are catalyzed and released by nitric oxide doped with selenium-containing selenocysteamine molecules.

S2 deposition of micro-nano particles released by nitric oxide catalysis

S3, deposition of Thrombin inhibitors

Example 7

The embodiment provides a vascular stent, which comprises the steps of adsorbing nitric oxide synthesized by polyphenol tannic acid and mono-selenocysteine on the surface of a 316L SS vascular stent to catalytically release micro-nano particles, and then depositing a bivalirudin thrombin inhibitor, wherein the preparation steps are as follows:

s1 preparation of nitrogen monoxide catalytic release micro-nano particles

Weighing tannic acid, dissolving in PBS buffer solution with pH of 8, and preparing into polyphenol tannic acid solution A with concentration of 1 mg/mL.

Weighing mono-selenocysteine molecules, dispersing in PBS buffer solution with the pH value of 8, and preparing into 0.1mg/mL mono-selenocysteine solution B.

And (3) mixing the solution A, the solution B and the solution C according to the mass ratio of 1: 1: 0, mixing, stirring at 40 ℃ or carrying out oscillation reaction for 24 hours; after the reaction is finished, separating to obtain micro-nano particles which are based on polyphenol tannic acid and contain selenium and sulfur and are catalyzed and released by nitric oxide doped with mono-selenocysteamine molecules.

S2 deposition of micro-nano particles released by nitric oxide catalysis

Weighing the nitric oxide catalytic release micro-nano particles obtained in the step S1, dispersing the nitric oxide catalytic release micro-nano particles into PBS buffer solution to prepare nitric oxide catalytic release micro-nano particle deposition solution with the concentration of 10mg/mL, soaking the vascular stent into the nitric oxide catalytic release micro-nano particle deposition solution, reacting for 12 hours at 25 ℃, and after the reaction is finished, fully rinsing and drying the vascular stent by using distilled water to obtain the vascular stent with the surface deposited with the nitric oxide catalytic release micro-nano particles.

S3, deposition of Thrombin inhibitors

Example 8

The embodiment provides a blood vessel stent, which comprises a step of adsorbing micro-nano particles which are synthesized by tannic acid and selenocysteine and contain selenium and nitrogen monoxide and are catalytically released on the surface of a 316L SS blood vessel stent, and a step of depositing a heparin thrombin inhibitor, wherein the preparation steps are as follows:

s1 preparation of nitrogen monoxide catalytic release micro-nano particles

And (3) mixing the solution A, the solution B and the solution C according to the mass ratio of 1: 1: 0, mixing, stirring at 40 ℃ or carrying out oscillation reaction for 24 hours; after the reaction is finished, separating to obtain the micro-nano particles which take polyphenol tannic acid as a structural basis and are catalyzed and released by nitric oxide doped with selenium-containing selenocysteamine molecules.

S2 deposition of micro-nano particles released by nitric oxide catalysis

S3, deposition of Thrombin inhibitors

Weighing heparin, dispersing the heparin into distilled water to prepare a heparin solution with the concentration of 1mg/mL, soaking the intravascular stent which is obtained in the step S2 and has the surface deposited with the nitric oxide catalytic release micro-nano particles into the heparin solution, depositing the intravascular stent for 2 hours at 37 ℃, and after the reaction is finished, fully rinsing the intravascular stent with distilled water and drying the intravascular stent to obtain the intravascular stent.

Example 9

The embodiment provides a vascular stent, which comprises the steps of grafting tannic acid and selenium-containing nitric oxide synthesized by cysteamine selenide on the surface of an aminated 316L SS-enriched vascular stent to catalyze and release micro-nano particles, and then depositing a bivalirudin thrombin inhibitor, wherein the preparation steps are as follows:

s1 preparation of nitric oxide catalytic release micro-nano particles

S2 deposition of micro-nano particles released by nitric oxide catalysis

Weighing the nitric oxide catalytic release micro-nano particles obtained in the step S1, dispersing the nitric oxide catalytic release micro-nano particles into PBS buffer solution with the pH value of 8 to prepare nitric oxide catalytic release micro-nano particle deposition solution with the concentration of 10mg/mL, soaking the intravascular stent into the nitric oxide catalytic release micro-nano particle deposition solution, reacting for 24 hours at 40 ℃, and after the reaction is finished, fully rinsing and drying the intravascular stent with the nitric oxide catalytic release micro-nano particles deposited on the surface.

S3, deposition of Thrombin inhibitors

Weighing bivalirudin, dispersing the bivalirudin in distilled water to prepare a bivalirudin solution with the concentration of 1mg/mL, soaking the vascular stent which is obtained in the step S2 and has the surface deposited with the nitric oxide to catalyze and release the micro-nano particles in the bivalirudin solution, depositing the vascular stent for 24 hours at 40 ℃, and after the reaction is finished, fully rinsing the vascular stent with distilled water and drying the vascular stent to obtain the bivalirudin preparation.

Comparative example 1

The comparative example provides a vascular stent, the preparation method of which is substantially the same as that of example 1, and the difference is that in the preparation of the nitric oxide catalytic release micro-nano particles, a solution A, a solution B and a solution C are mixed according to the mass ratio of 1: 0.0001: and (4) mixing.

Comparative example 2

The comparative example provides a vascular stent, the preparation method of which is substantially the same as that of example 1, and the difference is that in the preparation of the nitric oxide catalytic release micro-nano particles, a solution A, a solution B and a solution C are mixed according to the mass ratio of 1: 1: 101, mixing.

Comparative example 3

This comparative example provides a vascular stent which was prepared in substantially the same manner as in example 1, except that the thrombin inhibitor was deposited at 81 ℃. Subsequent characterization indicated that the deposited thrombin inhibitor bivalirudin lost thrombin inhibitory activity.

Test example 1

The nitric oxide catalytic release micro-nano particles prepared in the embodiment 1 are observed under an electron microscope to obtain results shown in fig. 1-5, and as can be seen from fig. 1, the nitric oxide catalytic release micro-nano particles prepared in the invention have the particle size in a nanometer scale and can be better deposited on a vascular stent. From fig. 2 to 5, it can be found that the nitrogen monoxide catalytic release micro-nano particles prepared in embodiment 1 of the present invention are uniformly doped with selenium in a compound skeleton formed by C, N, O elements, and have a function of catalytic release of nitrogen monoxide.

Test example 2

The nitric oxide catalytic release micro-nano particles prepared in the example 1 are placed in an NOA280i expiration nitric oxide determinator to be subjected to a nitric oxide catalytic release rate test, and a result shown in FIG. 6 is obtained, and as can be seen from FIG. 6, after the nitric oxide catalytic release micro-nano particles provided in the example are added for about 8min, the catalytic release rate of nitric oxide is slowly reduced to 0.0032 +/-0.0004 nmol/(ng & min) in a short time, and the release rate is maintained to continuously release nitric oxide.

Test example 3

The micro-nano particles for catalytic release of nitric oxide prepared in example 1 are coated with cationic dye Toluidine Blue (TBO), anionic dye acid orange (AO ii), hydrophobic drug Doxorubicin (DOX), synthetic polypeptide thrombin inhibitor Bivalirudin (BVLD), synthetic polypeptide antimicrobial peptide (AMP), protein Lysozyme (Lysozyme), protein Bovine Serum Albumin (BSA), and RNA extracted from human 3T3 cells by a particle solution and model molecule solution blending method, and the coating rate is detected, so that the results shown in fig. 7 are obtained. As can be seen from fig. 7, the micro-nano particles for catalytic release of nitric oxide prepared by the present invention can realize entrapment of various small molecules, polypeptides, proteins, RNAs, etc.

Test example 4

The nitric oxide release rate of the stent prepared in example 1 was measured by Griess method, and the results shown in fig. 8 were obtained. As can be seen from fig. 8, after the nitric oxide is deposited on the surface of the 316L SS intravascular stent to catalytically release the micro-Nano Particles (NPs), the release rate of nitric oxide is increased, and after Bivalirudin (BVLD) is deposited on the surface of the nitric oxide, the release rate of nitric oxide is not obviously changed, which indicates that the thrombin inhibitor bivalirudin is deposited on the surface of the intravascular stent on which the nitric oxide is deposited to catalytically release the micro-nano particles again, and the release of nitric oxide is not significantly affected.

Test example 5

The release rate of the stent prepared in example 1 was measured by uv-vis spectrophotometry, and the results shown in fig. 9 were obtained. As can be seen from fig. 9, the elution speed of bivalirudin is stabilized at 100 nmol/day, which indicates that the method provided by the present invention can realize continuous, stable and controllable elution of bivalirudin.

Test example 6

A blood coagulation analyzer is adopted to detect thrombin coagulation time of plasma after soaking treatment of 316L SS intravascular stent, aminated intravascular stent, stent deposited with nitric oxide catalytic release micro-nano particles and stent loaded with thrombin inhibitor bivalirudin deposited with nitric oxide catalytic release micro-nano particles, and results shown in figure 10 are obtained. As can be seen from fig. 10, the anti-coagulation effect of the intravascular stent obtained by depositing nitric oxide on the surface of the intravascular stent to catalytically release the micro-nano particle-loaded thrombin inhibitor is significantly improved.

Test example 7

The nitric oxide catalytic release micro-nano particle deposition solutions for preparing the vascular stent prepared in the embodiment 1 and the comparative example 1 are placed into a centrifuge for centrifugation, and the results shown in fig. 11-12 are obtained.

As can be seen from the figure, the sediment liquid in fig. 11 has a precipitate 101 at the bottom of the solution after centrifugation, and the sediment liquid in fig. 12 has no precipitate 101 after centrifugation, which indicates that no nitric oxide catalytic release micro-nano particles are generated in the mixing process of the solution a, the solution B and the solution C in comparative example 1, that is, the vascular stent prepared in comparative example 1 has no function of catalytically releasing nitric oxide.

Test example 8

By adopting the completely same preparation method of the embodiment 1 and the comparative example 2, the nitric oxide catalytic release micro-nano particles and the thrombin inhibitor are respectively deposited on the surfaces of two same 316L stainless steel sheets, and the size of the sheets is 10 multiplied by 0.05 mm.

Endothelial cells were cultured on 316L stainless steel sheets (blank control) of example 1, comparative example 2 and blank, respectively, and after 1 day, the cells were stained with a fluorescent dye and the number and state of the cells were observed in a fluorescent state to determine the activity of the cells. The results shown in FIGS. 13 to 15 were obtained.

As can be seen from the figure, the difference between the number of cells in fig. 14 and the number of cells in fig. 13 is not large, and the number of cells in fig. 15 is significantly reduced compared to fig. 13, which indicates that after the ratio of the solution A, B and the solution C is changed in comparative example 2, the obtained micro-nano particles with nitric oxide catalytic release have cell tissue toxicity and are not suitable for surface modification of vascular stents.

The CCK-8 kit is used for counting the number of endothelial cells on 316L stainless steel sheets in the images 13-15, and the result shown in the image 16 is obtained. Fig. 16 clearly shows that the number of cells in comparative example 2 is reduced by one time compared with example 3 and blank control, and further shows that after the proportion of solution A, B and C is changed in comparative example 2, the obtained micro-nano particles with nitric oxide catalytic release have cell tissue toxicity and are not suitable for surface modification of vascular stents.

In summary, the intravascular stent and the preparation method and application thereof provided by the invention have at least the following advantages:

1. the nitrogen monoxide catalytic release micro-nano particles obtained by using one or more of phenolic compounds, quinone compounds and ketone compounds as compound frameworks and doping the compounds with the nitrogen monoxide catalytic release function have the chemical structure stability at room temperature and normal pressure, active reaction functional groups on the surfaces can be grafted with other molecules or grafted to the surfaces of other materials, so that the drugs can be effectively loaded and released in a controlled manner, the controllable elution of the drugs is realized, the release time of the drugs is prolonged, and the slow release effect is improved.

2. By doping the compound with the nitric oxide catalytic release function in the compound skeleton, the finally obtained nitric oxide catalytic release micro-nano particles can catalytically release nitric oxide in the compound skeleton, so that the formation of thrombus is further inhibited.

3. According to the invention, the thrombin inhibitor is fixed on the surface of the blood vessel stent in an adsorption mode, and compared with a covalent grafting method, the adsorption and fixation of the thrombin inhibitor can prolong the elution time of the thrombin inhibitor and realize the controllable elution of the thrombin inhibitor.

The above is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and various modifications and changes will occur to those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A preparation method of a vascular stent is characterized by comprising the steps of sequentially depositing nitric oxide catalytic release micro-nano particles and thrombin inhibitor on the surface of the vascular stent, and carrying out surface modification on the vascular stent;

the nitric oxide catalytic release micro-nano particles comprise a compound framework and a compound doped in the compound framework and having a nitric oxide catalytic release function, wherein the compound framework comprises one or more of phenolic compounds, quinone compounds and ketone compounds;

the thrombin inhibitor is deposited in an immobilized manner by adsorption.

2. The preparation method according to claim 1, wherein the nitric oxide catalytic release micro-nano particles are obtained by mixing solution A, solution B and solution C and then reacting, and after the reaction is finished, performing solid-liquid separation;

the solution A is prepared by dissolving one or more of phenolic compounds, quinone compounds and ketone compounds in a dispersion medium; the solution B is prepared by dispersing a compound with a nitric oxide catalytic release function in a dispersion medium; the solution C is prepared by dissolving a compound containing copper ions in a dispersion medium;

preferably, the mixing ratio of the solution A, the solution B and the solution C is 1: 0.001 to 100: 0 to 100 parts; more preferably, the solution a, the solution B and the solution C are mixed in a ratio of 1: 0.001-10: 0 to 1;

preferably, the concentration of the solution A, the concentration of the solution B and the concentration of the solution C are all 0.001-100 mg/mL;

more preferably, the concentration of the solution A is 0.001-10 mg/mL, the concentration of the solution B is 0.001-1 mg/mL, and the concentration of the solution C is 0.001-0.1 mg/mL;

preferably, the reaction is a stirring or oscillation reaction, the reaction temperature is 0-80 ℃, and the reaction time is 0.01-72 hours; more preferably, the reaction temperature is 4-40 ℃, and the reaction time is 2-24 h.

3. The production method according to claim 1 or 2, wherein the phenolic compound comprises at least one of monophenol and derivatives thereof, catechol and derivatives thereof, pyrogallol and derivatives thereof, and phloroglucinol and derivatives thereof;

preferably, the quinone compound comprises at least one of benzoquinone and its derivatives, naphthoquinone and its derivatives, phenanthrenequinone and its derivatives, and anthraquinone and its derivatives;

preferably, the ketone compound comprises at least one of a flavonoid compound, a flavonol compound and a dihydroflavonoid compound;

preferably, the compound having the nitric oxide catalytic release function comprises ions and/or molecules having the nitric oxide catalytic release function;

preferably, the ions having nitric oxide catalytic release function comprise copper ion compounds or organic copper ions; more preferably, the copper ion is Cu⁺Or Cu²⁺；

4. The preparation method of claim 1, wherein the method for fixing nitric oxide catalytic release micro-nano particles on the surface of the vascular stent comprises the following steps: dispersing the nitric oxide catalytic release micro-nano particles in a deposition medium to obtain nitric oxide catalytic release micro-nano particle deposition solution, and then putting the intravascular stent into the nitric oxide catalytic release micro-nano particle deposition solution for deposition to obtain the intravascular stent with the nitric oxide catalytic release function;

preferably, the immobilization mode of the nitric oxide catalytic release micro-nano particle deposition comprises any one of adsorption, electrostatic interaction, covalent grafting or bridging anchoring;

preferably, the concentration of the nitric oxide catalyzed release micro-nano particle deposition solution is 0.001-100 mg/mL, and more preferably 0.05-10 mg/mL;

preferably, the deposition medium comprises one or more of distilled water with the pH of 4-12, PBS buffer solution, Tris-hydrochloric acid buffer solution, HEPES buffer solution, ethanol water solution, tetrahydrofuran water solution, acetone, benzene and derivatives thereof, dimethyl sulfoxide, dimethylformamide and acetonitrile;

the deposition temperature is 4-80 ℃, and the deposition time is 0.01-72 h; preferably, the deposition temperature is 4-40 ℃, and the reaction time is 0.5-24 h.

5. The method of claim 1, wherein the step of depositing the thrombin inhibitor on the surface of the stent comprises: the intravascular stent fixed with the nitric oxide catalytic release micro-nano particles is placed in thrombin inhibitor solution for deposition, so that the intravascular stent with the nitric oxide catalytic release function and the controllable elution of thrombin inhibitor is obtained;

the thrombin inhibitor solution is prepared by dispersing a thrombin inhibitor into a dispersion medium to prepare the thrombin inhibitor solution with the concentration of 0.001-100 mg/mL;

preferably, the concentration of the thrombin inhibitor solution is 0.001-10 mg/mL, more preferably 0.05-5 mg/mL;

preferably, the deposition temperature is 4-80 ℃, the deposition time is 0.01-72 h, more preferably, the deposition temperature is 4-37 ℃, and the reaction time is 0.5-24 h.

6. The method of claim 5, wherein the thrombin inhibitor comprises a direct or indirect thrombin inhibitor;

preferably, the direct thrombin inhibitor comprises one or more of dabigatran etexilate, bivalirudin, argatroban and recombinant hirudin;

7. The production method according to claim 2 or 5, the dispersion medium comprises one or more of distilled water with the pH value of 2-14, a barbital sodium buffer solution, a barbital sodium-hydrochloric acid buffer solution, a glycine-sodium hydroxide buffer solution, a glycine-hydrochloric acid buffer solution, a HEPES buffer solution, a phthalic acid-hydrochloric acid buffer solution, a potassium dihydrogen phosphate-sodium hydroxide buffer solution, a phosphate buffer solution, a citric acid-phosphoric acid buffer solution, a citric acid-sodium citrate buffer solution, a citric acid-sodium hydroxide-hydrochloric acid buffer solution, a PBS buffer solution, a borax-boric acid buffer solution, a borax-sodium hydroxide buffer solution, a TAE buffer solution, a sodium carbonate-sodium bicarbonate buffer solution, a TBST buffer solution, a TE buffer solution, a TEN buffer solution, a Tris-hydrochloric acid buffer solution and an acetic acid-sodium acetate buffer solution.

8. The preparation method according to claim 1, wherein the material of the vascular stent is metal or polymer;

9. A vascular stent prepared by the preparation method of any one of claims 1 to 8.

10. Use of the vascular stent of claim 9 in the preparation of a vascular repair article or an animal model of vascular repair.