CN110833631B - Preparation method of multifunctional intravascular stent - Google Patents
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- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/08—Materials for coatings
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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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/08—Materials for coatings
- A61L31/082—Inorganic materials
- A61L31/084—Carbon; Graphite
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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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/16—Biologically active materials, e.g. therapeutic substances
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- 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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- 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
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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
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/416—Anti-neoplastic or anti-proliferative or anti-restenosis or anti-angiogenic agents, e.g. paclitaxel, sirolimus
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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
- 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
- A61L2300/45—Mixtures of two or more drugs, e.g. synergistic mixtures
Abstract
The invention discloses a preparation method of a multifunctional intravascular stent, which comprises the following steps of firstly preparing a dopamine coating on the surface of the intravascular stent through dopamine autopolymerization; then loading heparin into the chitosan functionalized graphene oxide, and further fixing the heparin-loaded graphene oxide on the surface of the intravascular stent with the dopamine coating to obtain a heparin-loaded intravascular stent; and finally, immersing the vascular stent into rapamycin solution to fully adsorb and load rapamycin, thereby obtaining the multifunctional vascular stent. Due to the antibacterial action and the endothelial cell growth promoting action of chitosan, the anticoagulant action and the selective endothelial cell growth promoting action of heparin and the antiproliferative action of rapamycin, the vascular stent prepared by the method can inhibit clinical complications of stent implantation from multiple ways, thereby improving the application effect of the vascular stent.
Description
Technical Field
The invention relates to the field of medical instruments and biomaterials, in particular to a preparation method of a multifunctional intravascular stent.
Background
Cardiovascular diseases such as coronary heart disease seriously threaten the life health of human beings, and the vascular stent interventional therapy is used as a main means for treating the coronary heart disease and has the advantages of low operation cost, small wound, quick postoperative recovery and the like. However, due to the poor biocompatibility of the polymer coating in the traditional stent system, delayed healing of vascular endothelium caused by antiproliferative drugs and the like, the incidence of thrombus and restenosis in the stent still exists after the stent is implanted, and the life safety of a patient is threatened. The concrete points are as follows: (1) the polymer carrier has poor biocompatibility, is easy to cause inflammation and blood coagulation reaction, induces thrombosis and intimal hyperplasia, and leads to in-stent restenosis; (2) the healing of vascular endothelial injury caused by the antiproliferative drugs is delayed, and the incidence of late thrombosis and restenosis is increased; (3) the function is single, and the current drug-eluting vascular stent for clinical application is basically a single drug-loaded stent, and usually has good anti-proliferative performance, but has poor other performances.
Graphene Oxide (GO) is a carbon nano material containing various chemical functional groups, and has a huge application prospect in the fields of biomaterials and tissue engineering due to the huge specific surface area, good mechanical properties and biocompatibility. The carboxyl rich in GO has good chemical reactivity, and can be functionalized by means of chemical reaction of the carboxyl, non-covalent interaction of GO and other substances, electrostatic interaction or hydrogen bond formation and the like. The chitosan has good degradability and biocompatibility, and is widely applied to researches on biological materials and tissue engineering, and researches show that the chitosan has good cell compatibility and antibacterial performance, can obviously promote migration and proliferation of vascular endothelial cells, contains a large amount of amino groups in the structure, has a large amount of positive charges in a dilute acid solution, adopts the chitosan to functionalize GO, and can obviously improve the biocompatibility of GO and endow GO with the positive charge characteristic.
Heparin is a polysaccharide substance with excellent blood compatibility, and is widely applied to surface modification of blood-contacting biomaterials. Research shows that heparin can not only improve the blood compatibility of the material, but also promote the growth of endothelial cells to a certain extent, even selectively promote the growth of the endothelial cells, so that the heparin is loaded on the surface of the vascular stent, and the vascular stent is endowed with good blood compatibility and endothelial cell growth promotion performance through the controllable release of the heparin. On the other hand, heparin contains a large number of carboxyl groups and sulfonic groups, so that the heparin presents strong electronegativity, can be fixed on the surface of a positively charged material through electrostatic interaction and is used for a surface coating of a blood vessel stent, and meanwhile, the heparin can be loaded on the surface of GO through hydrophobic interaction with GO, so that the anticoagulation performance and the endothelial cell growth promotion performance of the material are improved.
Research shows that the neointimal formation caused by rupture of the intima of blood vessels after implantation of the blood vessel stent and further caused by migration and proliferation of smooth muscle cells is a main pathological process of restenosis in the stent. Rapamycin is a macrolide immunosuppressant, and blocks signal transduction through different cytokine receptors, blocks the progress of T lymphocytes and other cells from the G1 phase to the S phase, thereby exerting immunosuppressive effect and inhibiting smooth muscle cell proliferation, and is widely applied to research and application of drug-eluting vascular stents. At present, the rapamycin drug eluting stent applied clinically mainly adopts high molecules as carriers, has poor biocompatibility and can cause inflammatory reaction to a certain degree, thereby causing clinical complications.
The invention takes GO with huge specific surface area as a drug carrier, carries out chitosan functionalization on the GO, loads heparin with good anticoagulation performance into the GO, then fixes the GO on the surface of a vascular stent with a dopamine coating, and finally loads rapamycin into the surface coating of the vascular stent, thereby fully utilizing the good antibacterial performance of chitosan, the endothelial cell growth promotion type, the excellent anticoagulation performance of heparin, the selective endothelial cell growth promotion type and the antiproliferation performance of rapamycin, and endowing the vascular stent with excellent multifunctional characteristics.
Disclosure of Invention
The purpose of the invention is as follows: in order to overcome the defects in the prior art, the invention mainly aims to improve the antibacterial property, the anti-proliferation property and the anticoagulation property of the intravascular stent by preparing the multifunctional bioactive coating on the surface of the intravascular stent.
The technical scheme is as follows: in order to solve the technical problems, the invention provides a preparation method of a multifunctional intravascular stent, which comprises the steps of firstly preparing a dopamine coating on the surface of the intravascular stent through dopamine autopolymerization; then loading heparin into the chitosan functionalized graphene oxide, and further fixing the chitosan functionalized graphene oxide on the surface of the intravascular stent with the dopamine coating to obtain the heparin-loaded intravascular stent; and finally, immersing the vascular stent into rapamycin solution for sufficient adsorption to obtain the multifunctional vascular stent.
The preparation method of the dopamine coating comprises the steps of immersing the blood vessel stent which is fully cleaned into 0.1-1 mg/ml dopamine solution with the pH value of 8.0-8.5, reacting at room temperature for 24 hours, fully cleaning and drying to obtain the blood vessel stent with the polydopamine coating.
The preparation method of the chitosan functionalized graphene oxide comprises the steps of ultrasonically dispersing graphene oxide and chitosan in MES buffer solution with the pH = 5-6, adding 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide and N-hydroxysuccinimide solution, carrying out oscillation reaction for 4, then repeatedly centrifuging and washing to remove unreacted substances, and thus obtaining the chitosan functionalized graphene oxide. The concentration of graphene oxide and chitosan is 1-5mg/ml, and the concentration of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide and N-hydroxysuccinimide is 5-10 mM.
The preparation method of the heparin-loaded graphene oxide comprises the steps of fully dispersing the obtained chitosan functionalized graphene oxide in a buffer solution with the pH value of 7.4, fully blending a heparin solution with the same concentration and volume, and repeatedly centrifuging and washing to obtain the heparin-loaded graphene oxide. The concentration of the chitosan functionalized graphene oxide and heparin is 1-5 mg/ml.
The preparation method of the heparin-loaded intravascular stent comprises the steps of fully dispersing the heparin-loaded graphene oxide obtained in the claim 4, immersing the intravascular stent with the dopamine coating into a dispersion liquid, and fully reacting for 4 hours to obtain the intravascular stent with the heparin-loaded graphene oxide coating on the surface. The concentration of the heparin-loaded graphene oxide is 1-5 mg/ml.
The method for loading rapamycin on the surface of the blood vessel stent comprises the steps of immersing the blood vessel stent carrying heparin into rapamycin solution with the concentration of 1-10mg/ml, fully adsorbing for 4-12 hours, and drying to obtain the multifunctional blood vessel stent.
Has the advantages that: compared with the prior art, the invention has the following advantages:
(1) the surface coating of the stent does not need a traditional high-molecular drug carrier, so that clinical side reactions such as inflammatory reaction and the like caused by the implantation of traditional high molecules are avoided;
(2) the chitosan functionalized graphene oxide with huge specific surface area is used as a carrier, and heparin with excellent anticoagulation performance and rapamycin with excellent anti-proliferative performance are loaded, so that two drugs are loaded simultaneously, and more drugs can be loaded due to the huge specific surface area of the graphene oxide, and the long-term application effect of the graphene oxide is improved.
(3) The intravascular stent prepared by the invention has good antibacterial property, excellent anticoagulation property, good endothelial cell growth promotion and anti-proliferation property, and endows the intravascular stent with excellent multifunctional property.
(4) The technical method provided by the invention can load various bioactive molecules and endow the coating with different biological properties; meanwhile, the method only needs to be carried out in solution, and has the advantages of high deposition speed, economy, easy implementation, no need of special equipment and the like.
Drawings
FIG. 1 shows the growth of endothelial cells on the surface of multifunctional vascular stent.
FIG. 2 shows endothelialization of the multifunctional vascular stent in vivo for one week.
Detailed Description
The present invention is specifically illustrated by the following examples, which are only used to more clearly illustrate the technical solutions of the present invention, and the protection scope of the present invention is not limited thereby.
Example 1
Preparation of multifunctional vascular stent
(1) Preparation of dopamine coated vascular stent
And ultrasonically cleaning the vascular stent for 10 minutes by using acetone, ethanol and distilled water in sequence, drying, immersing into 0.2mg/ml dopamine solution with the pH value of 8.0-8.5, reacting at room temperature for 24 hours, and fully cleaning and drying to obtain the vascular stent with the polydopamine coating.
(2) Preparation of chitosan functionalized graphene oxide
Respectively ultrasonically dispersing graphene oxide and chitosan in MES (MES) buffer solution with the pH = 5-6 to respectively obtain 1mg/ml dispersion liquid, respectively and fully mixing 5ml graphene oxide and chitosan solution, respectively adding 1ml 5mM 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide and N-hydroxysuccinimide solution, and repeatedly centrifuging and washing after oscillation reaction for 4 to remove unreacted substances to obtain the functionalized graphene oxide of chitosan.
(3) Preparation of heparin-loaded vascular stents
Fully dispersing the chitosan functionalized graphene oxide obtained in the step (2) in a buffer solution with the pH value of 7.4 to obtain 1mg/ml dispersion liquid, adding an isovolumetric heparin solution of 1mg/ml, fully blending, and repeatedly centrifuging and washing to obtain the heparin-loaded graphene oxide. Re-dispersing the heparin-loaded graphene oxide to obtain 1mg/ml dispersion liquid, immersing the intravascular stent obtained in the step (1) into the dispersion liquid, and fully reacting for 4 hours to obtain the intravascular stent with the surface provided with the heparin-loaded graphene oxide coating.
(4) Preparation of rapamycin-loaded vascular stents
And (4) immersing the heparin-loaded vascular stent obtained in the step (3) into a rapamycin solution of 1mg/ml, fully adsorbing for 4 hours, and drying to obtain the multifunctional vascular stent.
Example 2
Preparation of multifunctional vascular stent
(1) Preparation of dopamine coated vascular stent
And ultrasonically cleaning the vascular stent for 10 minutes by using acetone, ethanol and distilled water in sequence, drying, immersing into 0.5mg/ml dopamine solution with the pH value of 8.0-8.5, reacting at room temperature for 24 hours, and fully cleaning and drying to obtain the vascular stent with the polydopamine coating.
(2) Preparation of chitosan functionalized graphene oxide
Respectively ultrasonically dispersing graphene oxide and chitosan in MES (MES) buffer solution with the pH = 5-6 to obtain 3mg/ml dispersion liquid, respectively and fully mixing 5ml graphene oxide and chitosan solution, respectively adding 1ml 5mM 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide and N-hydroxysuccinimide solution, and repeatedly centrifuging and washing after oscillation reaction for 4 to remove unreacted substances to obtain the functionalized graphene oxide of chitosan.
(3) Preparation of heparin-loaded vascular stents
Fully dispersing the chitosan functionalized graphene oxide obtained in the step (2) in a buffer solution with the pH value of 7.4 to obtain a dispersion liquid of 3mg/ml, adding a heparin solution with the same volume as that of the dispersion liquid of 3mg/ml, fully blending, and repeatedly centrifuging and washing to obtain the heparin-loaded graphene oxide. Re-dispersing the heparin-loaded graphene oxide to obtain 3mg/ml dispersion liquid, immersing the intravascular stent obtained in the step (1) into the dispersion liquid, and fully reacting for 4 hours to obtain the intravascular stent with the surface provided with the heparin-loaded graphene oxide coating.
(4) Preparation of rapamycin-loaded vascular stents
And (4) immersing the heparin-loaded vascular stent obtained in the step (3) into a rapamycin solution of 5mg/ml, fully adsorbing for 12 hours, and drying to obtain the multifunctional vascular stent.
Example 3
Preparation of multifunctional vascular stent
(1) Preparation of dopamine coated vascular stent
And ultrasonically cleaning the vascular stent for 10 minutes by using acetone, ethanol and distilled water in sequence, drying, immersing into 1mg/ml dopamine solution with the pH of 8.0-8.5, reacting at room temperature for 24 hours, and fully cleaning and drying to obtain the vascular stent with the polydopamine coating.
(2) Preparation of chitosan functionalized graphene oxide
Respectively ultrasonically dispersing graphene oxide and chitosan in MES (MES) buffer solution with the pH = 5-6 to respectively obtain 5mg/ml dispersion liquid, respectively and fully mixing 5ml graphene oxide and chitosan solution, respectively adding 1ml 10mM 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide and N-hydroxysuccinimide solution, and repeatedly centrifuging and washing after oscillation reaction for 4 to remove unreacted substances to obtain the functionalized graphene oxide of chitosan.
(3) Preparation of heparin-loaded vascular stents
Fully dispersing the chitosan functionalized graphene oxide obtained in the step (2) in a buffer solution with the pH value of 7.4 to obtain 5mg/ml dispersion liquid, adding an isovolumetric heparin solution of 5mg/ml, fully blending, and repeatedly centrifuging and washing to obtain the heparin-loaded graphene oxide. Re-dispersing the heparin-loaded graphene oxide to obtain 5mg/ml dispersion liquid, immersing the intravascular stent obtained in the step (1) into the dispersion liquid, and fully reacting for 4 hours to obtain the intravascular stent with the surface provided with the heparin-loaded graphene oxide coating.
(4) Preparation of rapamycin-loaded vascular stents
And (4) immersing the heparin-loaded vascular stent obtained in the step (3) into a rapamycin solution of 10mg/ml, fully adsorbing for 8 hours, and drying to obtain the multifunctional vascular stent.
The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (3)
1. A preparation method of a multifunctional blood vessel support comprises the steps of firstly preparing a polydopamine coating on the surface of the blood vessel support; then loading heparin into the chitosan functionalized graphene oxide, and further fixing the heparin-loaded graphene oxide on the surface of the blood vessel stent with the polydopamine coating to obtain a heparin-loaded blood vessel stent; finally, immersing the vascular stent into rapamycin solution to fully adsorb and load the antiproliferative drugs to obtain a multifunctional vascular stent; the preparation method of the polydopamine coating comprises the steps of immersing the blood vessel stent which is fully cleaned into 0.1-1 mg/ml dopamine solution with the pH value of 8.0-8.5, carrying out self-polymerization reaction at room temperature for 24 hours, and fully cleaning and drying to obtain the blood vessel stent with the polydopamine coating; fully dispersing the obtained chitosan functionalized graphene oxide in a buffer solution with the pH value of 7.4, fully blending a heparin solution with the same concentration and volume, repeatedly centrifuging and washing to obtain the heparin-loaded graphene oxide, wherein the concentrations of the chitosan functionalized graphene oxide and the heparin are 1-5 mg/ml; the preparation method of the heparin-loaded intravascular stent comprises the steps of fully dispersing the obtained heparin-loaded graphene oxide, immersing the intravascular stent with the polydopamine coating into dispersion liquid, and fully reacting for 4 hours to obtain the intravascular stent with the surface loaded with heparin, wherein the concentration of the heparin-loaded graphene oxide is 1-5 mg/ml.
2. The preparation method of the multifunctional vascular stent as claimed in claim 1, wherein the preparation method of the chitosan functionalized graphene oxide comprises the steps of ultrasonically dispersing graphene oxide and chitosan in MES buffer solution with pH of 5-6, adding 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide and N-hydroxysuccinimide solution, and repeatedly centrifuging and washing to remove unreacted substances after oscillation reaction for 4 hours to obtain the chitosan functionalized graphene oxide; the concentration of graphene oxide and chitosan is 1-5mg/ml, and the concentration of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide and N-hydroxysuccinimide is 5-10 mM.
3. The preparation method of the multifunctional vascular stent as claimed in claim 1, wherein the rapamycin is loaded on the surface of the vascular stent by immersing the obtained vascular stent into a rapamycin solution of 1-10mg/ml, fully adsorbing for 4-12 hours, and drying to obtain the multifunctional vascular stent.
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| CN112870437B (en) * | 2021-01-18 | 2023-04-18 | 成都鼎峰前瞻科技有限公司 | Functional material with anticoagulation, anti-hyperplasia and endothelialization promotion functions, and preparation method and application thereof |
| CN114246992B (en) * | 2021-12-28 | 2022-12-20 | 湖南华翔医疗科技有限公司 | Degradable intravascular stent with slow-release drug coating and preparation method thereof |
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