CN101357241B - CD34 antibody or CD133 antibody surface orientation fixing method of titanium and titanium alloy cardiovascular implantation device - Google Patents
CD34 antibody or CD133 antibody surface orientation fixing method of titanium and titanium alloy cardiovascular implantation device Download PDFInfo
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- CN101357241B CN101357241B CN2008100460480A CN200810046048A CN101357241B CN 101357241 B CN101357241 B CN 101357241B CN 2008100460480 A CN2008100460480 A CN 2008100460480A CN 200810046048 A CN200810046048 A CN 200810046048A CN 101357241 B CN101357241 B CN 101357241B
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Abstract
The invention discloses a method which adopts the surface of a titanium and titanium alloy cardiovascular implantation device to fix CD34 antibody or CD133 antibody directionally, and comprises the following main steps: A. activation treatment is carried out, so as to form hydroxyl on the surface; B. a layer of avidin is adsorbed on the activation surface of the step A; C. a layer of biotinylatedprotein A is adsorbed by the avidin layer; D. on the surface of the protein A layer obtained, a layer of specific antibodies of the membrane protein of endothelial progenitor cells- CD34 antibodies or CD133 antibodies are fixed directionally, so the titanium and titanium alloy cardiovascular implantation device can capture the endothelial progenitor cells in blood after being implanted into the human body, thereby leading to rapid endothelialization of the implantation device surface and leading the implantation device to have excellent anticoagulant function and restenosis occurring resistance function.
Description
Affiliated technical field
The present invention relates to the bioid method of modifying of titanium or titanium alloy cardiovascular implantation device.
Background technology
Cardiovascular system diseases is one of healthy important diseases of harm humans, and the artificial titanium or titanium alloy cardiovascular implantation device (intravascular stent or cardiac valve) of implanting is the effective ways of treating cardiovascular disease.The whole world has about 2,000,000 coronary heart disease patients of surpassing need carry out percutaneous transluminal coronary angioplasty every year at present, and wherein 70% needs the implantable intravascular support.The patient that China accepted the intravascular stent implanted treatment in 2006 also exceedes 100,000 person-times.China's valvular heart disease accounts for cardiac 30%, also has every year 100000 above critical patients must implement artificial Cardiac Valve Replacement.
Sigwart in 1987 adopts titanium or titanium alloy intracoronary stent (BMS) to implant coronary artery first, but has 20%～30% case that in-stent restenosis (ISR, promptly in the support because the smooth muscle cell hyper-proliferative causes the tube chamber narrow once again) can take place.Main clinically at present the application through using coating stent of medicine that the restenosis incidence rate is reduced to below 10%, coating stent of medicine mainly contains two kinds: rapamycins coating support and polymer are the paclitaxel coating bracket on basis.Though coating stent of medicine can reduce the restenosis incidence rate, because the medicine in the coating when suppressing smooth muscle cell proliferation, has also suppressed the propagation of endotheliocyte, make support exposed, cause inflammation, blood coagulation even form thrombosis.
China is annual owing to the valve pathological changes seriously needs to implement the patient that the artificial heart valve replacement is performed the operation at present, about 200,000 examples, and these patient's majorities are between twenty and fifty, change lobe as can not in time performing the operation, and will cause immeasurable loss to society.Cardiac valve prosthesis commonly used at present is divided into mechanical valve prosthesis and biovalve two big classes.The former adopts the titanium or titanium alloy material to process, because of its preferably durability be widely applied, but use patient's postoperative of mechanical prosthetic valve must take anticoagulant such as warfarin for a long time, anti-tampon.Bioprosthetic valve is because problem of aging possibly need the replacement that undergos surgery once more after certain after surgery year.If can make mechanical valve prosthesis surface form the complete endothelial layer of one deck, the probability that makes blood coagulation is reduced greatly, promptly can reduce or no longer take anticoagulant, also need change valve hardly simultaneously, reduce patient's medical expense greatly.
Endothelial progenitor cells in the key that works of endothelial progenitor cells in impaired cardiovascular endothelialization, blood circulation can be by chemotactic to cardiovascular damage location, and its differentiation, multiplication potentiality can make the structure function of damage endothelium repair fast.Form similar natural cardiovascular inner membrance on the cardiovascular implantation device surface; Be that the implant surface endothelialization is considered to the final fundamental way of implanting back thrombosis problem and improving long-term patency rate that solves, the quick endothelialization of damage inner membrance is important measures that do not solve neointimal hyperplasia and restenosis.
Summary of the invention
The object of the invention just provides a kind of surface orientation fixation of C D34 antibody of titanium or titanium alloy cardiovascular implantation device or the method for CD133 antibody; This kind method can be fixed on CD34 antibody or CD133 antibody orientation the surface of titanium or titanium alloy cardiovascular implantation device; Can catch endothelial progenitor cells in the blood after making titanium or titanium alloy cardiovascular implantation device implant into body; Induce the quick endothelialization of implanting device, thereby make implanting device have good anticoagulation function and the function that suppresses the restenosis generation.
Be to realize that above-mentioned purpose, the technical scheme that the present invention taked are,
A kind of surface orientation fixation of C D34 antibody of titanium or titanium alloy cardiovascular implantation device or the method for CD133 antibody, its step is following:
A, activation processing
The titanium or titanium alloy cardiovascular implantation device was soaked 1-24 hour in 40-80 ℃ 0.5-5mol/L NaOH solution, take out and to be placed in the 60-90 ℃ of deionized water insulation 1-24 hour, again at 30-90 ℃ air drying;
Perhaps the titanium or titanium alloy cardiovascular implantation device was soaked 1 hour in the orthophosphoric acid solution of 0.5-5mol/L, clean drying;
Fixing of B, Avidin:
A is gone on foot implanting device after the activation processing immersed in normal saline that Avidin concentration is 0.1-5mg/ml or the phosphate buffer 1-12 hour, take out the back with cleaning drying;
Perhaps A is gone on foot implanting device after the activation processing behind deionized water, acetone, toluene rinsing successively; Placing the toluene that contains 1%-10%3-aminopropyl three oxygen ethylsilane to be heated to 60 ℃ refluxed 4 hours; Naturally cool to room temperature preservation 12h, take out the back and clean the final vacuum drying successively with toluene, ethanol, deionized water; Again its immersion is contained the 0.1-5mg/ml Avidin and contain in 2-(N-morphine quinoline) the ethyl sulfonic acid solution of N-hydroxy-succinamide and 0.1mg/ml of 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide, 0.25mg/ml of 1-10mg/ml and react 1-4h; Clean drying;
C, biotinylated protein A's is fixing:
Implanting device after B step handled immerses in the albumin solution of 0.1%-5%, and incubated at room 5-40 minute, again its immersion is contained in normal saline or the phosphate buffer of biotinylated protein A of 0.01-5mg/ml 1-12 hour, take out the back and clean drying;
D: the orientation of antibody is fixed:
The absorption that C step is obtained the implanting device of biotinylated protein A immersed in the albumin solution of 0.1%-5% 5-40 minute; Took out in the normal saline that immerses the CD34 antibody that contains 0.01-1mg/ml or CD133 antibody in the back or the phosphate buffer 1-12 hour; Take out the back and clean drying.
Chemical reaction process, mechanism that the present invention prepares in the process are:
Through the immersion of NaOH or orthophosphoric acid solution under the A step condition, make the surface of titanium or titanium alloy cardiovascular implantation device form loose structure and have a large amount of hydrophilic hydroxyls.
Pass through silane coupler at B in step again---the ethoxy group in the 3-aminopropyl three oxygen ethylsilane and the hydroxyl generation covalent bond on implanting device surface; Form the silane coupler monolayer at apparatus surface; Under 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide catalysts effect, the amino of silane coupler monolayer and the carboxyl of Avidin react again, and the fixing from the teeth outwards Avidin layer that forms.
Then combine with Avidin electric charge adsorption through albumin earlier in the C step; Displace the albumin that adsorbs on the Avidin by the biotin group among the biotinylated protein A (more being prone to combine) competition again, form the biotin group specific bond of Avidin and biotinylated protein A with Avidin than albumin.
To be that c-terminus is single-minded combine for antibody CD34 in D step in the fixed biotinylated protein A in implanting device surface and the soaking solution or the not function section of CD133; Thereby the functional domain end of antibody is outwards arranged away from the surface of solids; The orientation of realization antibody is fixed, in order to catch the endothelial progenitor cells in the blood circulation.
Compared with prior art, the invention has the beneficial effects as follows:
The hydroxyl that obtains through surface active and the coupling covalent bond Avidin of silane coupler are by Avidin and biotinylated protein A specific bond; Biotinylated protein A combines with the not function terminal specific of antibody; And its functional domain end is outwards arranged away from the surface of solids; Finally be implemented in the implanting device orientation and construct endothelial progenitor cells specific recognition surface, effectively catch endothelial progenitor cells in the blood, induce the quick endothelialization of implanting device.Because the endothelial progenitor cells surface of cell membrane height in the blood circulation is expressed CD34 or CD133 memebrane protein (belonging to antigen); Therefore; The present invention is employed in device materials function of surface end outside directed fixation of C D34 antibody or CD133 antibody, and the activity of having preserved antibody has to greatest extent improved the efficient of catching endothelial progenitor cells in the blood; Induce the quick endothelialization on implanting device surface, thereby make implanting device have good anticoagulation function and the function that suppresses the restenosis generation.
The experiment proof is with the titanium or titanium alloy cardiovascular of the inventive method behind surface orientation fixation of C D34 antibody or CD133 antibody (also promptly using the inventive method modification); Effective capturing endothelial ancestral cell; Induce implanting device surface endothelialization fast, have useful anticoagulation function.
The said A step is with NaOH immersion, insulation and dried implanting device, fixes with carrying out the Avidin that B goes on foot after irradiation under ultraviolet ray 1-30 minute again.Irradiation under ultraviolet ray can make the better effects if of activation processing.
The cleaning of above-mentioned A in the step is that deionized water for ultrasonic is cleaned, and be dry for nitrogen dries up, and B, C, the cleaning of D in the step are normal saline or phosphate buffer rinsing, and drying dries up for nitrogen.These cleaning ways make that cleaning is cleaner, and nitrogen dries up the activity that then can better keep active substance.
Description of drawings
Below in conjunction with accompanying drawing and embodiment method of the present invention is done further detailed explanation.
Fig. 1 is the optical microscope photograph of the experiment of the implanting device capturing endothelial ancestral cell after the modification of the present invention.
Fig. 2 is the optical microscope photograph of unmodified implanting device capturing endothelial ancestral cell experiment.
Fig. 3 is with the implanting device after the modification of the present invention, and the femoral artery of implanting adult dog is interior after 2 hours, takes out the electron scanning micrograph that obtains.
The unmodified implanting device of Fig. 4, the femoral artery of implanting adult dog is interior after 2 hours, takes out the electron scanning micrograph that obtains.
The specific embodiment
Embodiment 1
1, a kind of method of surface orientation fixation of C D34 antibody of titanium alloy cardiovascular implantation device, its step is following:
A, activation processing
The titanium alloy cardiovascular implantation device was soaked 1 hour in 40 ℃ 0.5mol/L NaOH solution, take out and to be placed in 60 ℃ of deionized waters insulation 1 hour, again at 30 ℃ air drying; With irradiation under ultraviolet ray 1 minute.
Fixing of B, Avidin:
A is gone on foot implanting device after the activation processing immersed in the normal saline that Avidin concentration is 0.1mg/ml 1 hour, take out the back and uses the normal saline rinsing, nitrogen dries up;
C, biotinylated protein A's is fixing:
Implanting device after B step handled immerses in 0.1% the albumin solution, and incubated at room 5 minutes contains in the normal saline of biotinylated protein A of 0.01mg/ml 1 hour with its immersion again, takes out back normal saline rinsing, and nitrogen dries up;
D: the orientation of antibody is fixed:
The absorption that C step is obtained the implanting device of biotinylated protein A immersed in 0.1% the albumin solution 5 minutes, took out in the normal saline that immerses the CD34 antibody that contains 0.01mg/ml in the back 1 hour, take out back normal saline rinsing, nitrogen dries up.
Embodiment 2
1, a kind of method of surface orientation fixation of C D34 antibody of titanium cardiovascular implantation device, its step is following:
A, activation processing
The titanium cardiovascular implantation device was soaked 24 hours in 80 ℃ 5mol/L NaOH solution, take out to be placed in 90 ℃ of deionized waters and be incubated 24h, again behind 90 ℃ air drying, with irradiation under ultraviolet ray 30 minutes.
Fixing of B, Avidin:
A is gone on foot implanting device after the activation processing immersed in the phosphate buffer that Avidin concentration is 5.0mg/ml 12 hours, take out the back and uses the phosphate buffer rinsing, nitrogen dries up;
C, biotinylated protein A's is fixing:
Implanting device after B step handled immerses in 5% the albumin solution, and incubated at room 40 minutes contains in the phosphate buffer of biotinylated protein A of 5mg/ml 12 hours with its immersion again, takes out back phosphate buffer rinsing, and nitrogen dries up;
D: the orientation of antibody is fixed:
The absorption that C step is obtained the implanting device of biotinylated protein A immersed in 5% the albumin solution 40 minutes, take out the back and immersed in the normal saline that contains 1mg/ml CD34 antibody or the phosphate buffer 12 hours, take out back phosphate buffer rinsing, nitrogen dries up.
Embodiment 3
1, a kind of method of surface orientation fixation of C D133 antibody of titanium alloy cardiovascular implantation device, its step is following:
A, activation processing
The titanium alloy cardiovascular implantation device was soaked 12 hours in 70 ℃ 3mol/L NaOH solution, take out and to be placed in 70 ℃ of deionized waters insulation 20 hours, again behind 80 ℃ air drying, with irradiation under ultraviolet ray 20 minutes.
Fixing of B, Avidin:
A is gone on foot implanting device after the activation processing immersed in the phosphate buffer that Avidin concentration is 1.0mg/ml 6 hours, take out the back and uses the phosphate buffer rinsing, nitrogen dries up;
C, biotinylated protein A's is fixing:
Implanting device after B step handled immerses in 1% the albumin solution, and incubated at room 40 minutes contains in the phosphate buffer of biotinylated protein A of 0.1mg/ml 12 hours with its immersion again, takes out back phosphate buffer rinsing, and nitrogen dries up;
D: the orientation of antibody is fixed:
The absorption that C step is obtained the implanting device of biotinylated protein A immersed in 1% the albumin solution 40 minutes, took out in the phosphate buffer that immerses the CD133 antibody that contains 0.1mg/ml in the back 12 hours, take out back phosphate buffer rinsing, nitrogen dries up.
Embodiment 4
1, a kind of method of surface orientation fixation of C D133 antibody of titanium cardiovascular implantation device, its step is following:
A, activation processing
The titanium cardiovascular implantation device was soaked 3 hours in the orthophosphoric acid solution of 5mol/L, and deionized water for ultrasonic is cleaned, and nitrogen dries up;
Fixing of B, Avidin:
A is gone on foot implanting device after the activation processing behind deionized water, acetone, toluene rinsing successively; Placing the toluene that contains 10%3-aminopropyl three oxygen ethylsilane to be heated to 60 ℃ refluxed 4 hours; Naturally cool to room temperature preservation 12h, take out the back and clean the final vacuum drying successively with toluene, ethanol, deionized water; Again its immersion is contained the 5mg/ml Avidin and contain in 2-(N-morphine quinoline) the ethyl sulfonic acid solution of N-hydroxy-succinamide and 0.1mg/ml of 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide, 0.25mg/ml of 10mg/ml and react 4h; Clean drying;
C, biotinylated protein A's is fixing:
Implanting device after B step handled immerses in 5% the albumin solution, and incubated at room 40 minutes contains in the phosphate buffer of biotinylated protein A of 5mg/ml 12 hours with its immersion again, takes out back phosphate buffer rinsing, and nitrogen dries up;
D: the orientation of antibody is fixed:
The absorption that C step is obtained the implanting device of biotinylated protein A immersed in 5% the albumin solution 40 minutes, took out in the phosphate buffer that immerses the CD133 antibody that contains 1mg/ml in the back 12 hours, take out back phosphate buffer rinsing, nitrogen dries up.
Embodiment 5
1, a kind of method of surface orientation CD133 antibody of titanium alloy cardiovascular implantation device, its step is following:
The titanium cardiovascular implantation device was soaked 2 hours in the orthophosphoric acid solution of 3mol/L, and deionized water for ultrasonic is cleaned, and nitrogen dries up;
Fixing of B, Avidin:
A is gone on foot implanting device after the activation processing behind deionized water, acetone, toluene rinsing successively; Placing the toluene that contains 5%3-aminopropyl three oxygen ethylsilane to be heated to 60 ℃ refluxed 4 hours; Naturally cool to room temperature preservation 12h, take out the back and clean the final vacuum drying successively with toluene, ethanol, deionized water; Again its immersion is contained the 0.5mg/ml Avidin and contain in 2-(N-morphine quinoline) the ethyl sulfonic acid solution of N-hydroxy-succinamide and 0.1mg/ml of 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide, 0.25mg/ml of 6mg/ml and react 2h; The normal saline rinsing, nitrogen dries up.
C, biotinylated protein A's is fixing:
Implanting device after B step handled immerses in 1% the albumin solution, and incubated at room 20 minutes contains in the normal saline of biotinylated protein A of 0.1mg/ml 5 hours with its immersion again, takes out back normal saline rinsing, and nitrogen dries up;
D: the orientation of antibody is fixed:
The absorption that C step is obtained the implanting device of biotinylated protein A immersed in 1% the albumin solution 20 minutes, took out in the normal saline that immerses the CD133 antibody that contains 0.05mg/ml in the back 5 hours, take out back normal saline rinsing, nitrogen dries up.
Embodiment 6
1, a kind of method of surface orientation fixation of C D34 antibody of titanium alloy cardiovascular implantation device, its step is following:
A, activation processing
The titanium alloy cardiovascular implantation device was soaked 0.5 hour in the orthophosphoric acid solution of 0.5mol/L, and deionized water for ultrasonic is cleaned, and nitrogen dries up;
Fixing of B, Avidin:
A is gone on foot implanting device after the activation processing immersed in the phosphate buffer that Avidin concentration is 1.0mg/ml 12 hours, take out the back and uses the phosphate buffer rinsing, nitrogen dries up;
C, biotinylated protein A's is fixing:
Implanting device after B step handled immerses in 0.1% the albumin solution, and incubated at room 5 minutes contains in the normal saline of biotinylated protein A of 0.01mg/ml 1 hour with its immersion again, takes out the back and cleans drying;
D: the orientation of antibody is fixed:
The absorption that C step is obtained the implanting device of biotinylated protein A immersed in 0.1% the albumin solution 20 minutes, took out in the normal saline that immerses the CD34 antibody that contains 0.01mg/ml in the back 1 hour, take out the back and clean drying.
Embodiment 7
1, a kind of method of surface orientation fixation of C D133 antibody of titanium alloy cardiovascular implantation device, its step is following:
A, activation processing
The titanium alloy cardiovascular implantation device was soaked 12 hours in 65 ℃ 2.5mol/L NaOH solution, take out and to be placed in 70 ℃ of deionized waters insulation 16 hours, again at 70 ℃ air drying.
Fixing of B, Avidin:
A is gone on foot implanting device after the activation processing behind deionized water, acetone, toluene rinsing successively; Placing the toluene that contains 1%3-aminopropyl three oxygen ethylsilane to be heated to 60 ℃ refluxed 4 hours; Naturally cool to room temperature preservation 12h, take out the back and clean the final vacuum drying successively with toluene, ethanol, deionized water; Again its immersion is contained the 0.1mg/ml Avidin and contain in 2-(N-morphine quinoline) the ethyl sulfonic acid solution of N-hydroxy-succinamide and 0.1mg/ml of 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide, 0.25mg/ml of 1mg/ml and react 1h; Clean drying;
C, biotinylated protein A's is fixing:
Implanting device after B step handled immerses in 1% the albumin solution, and incubated at room 40 minutes contains in the phosphate buffer of biotinylated protein A of 0.1mg/ml 12 hours with its immersion again, takes out back phosphate buffer rinsing, and nitrogen dries up;
D: the orientation of antibody is fixed:
The absorption that C step is obtained the implanting device of biotinylated protein A immersed in 1% the albumin solution 40 minutes, took out in the phosphate buffer that immerses the CD133 antibody that contains 0.1mg/ml in the back 12 hours, take out back phosphate buffer rinsing, nitrogen dries up.
Following experimental result proves that the cardiovascular implantation device after the inventive method modification can be caught endothelial progenitor cells in the blood, induces the quick endothelialization of implanting device:
Test under the moving chamber simulation of an extracorporeal flow blood flow condition blood vessel implanting device surface to the capture ability of endothelial progenitor cells.
Cardiovascular implantation device after the inventive method modification and unmodified implanting device are placed in the flat flow chamber endothelial progenitor cells suspension (density 1 * 10 of 50ml respectively
6/ ml) be fed into the flow cavity system, regulate the flow rate of liquid in the flow cavity, make the shearing force size be about 1.0Pa, similar arteriolar shearing force.Flat flow chamber is at CO
2Incubator (37C, 5%CO
2) hatch 24h.Take out washing gently in PBS then.Observe the cell quantity of different materials surface adhesion.Fig. 1 is the optical microscope photograph of the experiment of the implanting device capturing endothelial ancestral cell after the modification of the present invention, and Fig. 2 is the optical microscope photograph of unmodified implanting device capturing endothelial ancestral cell experiment.Fig. 1, Fig. 2 show that the implanting device coating after the modification of the present invention catches the quantity of the endothelial progenitor cells in the dynamic circulation liquid and increase greatly.
Experiment disome IT endothelial progenitor cells, the endothelialization experiment of induced material surface.
Cardiovascular implantation device after the inventive method modification and unmodified implanting device are implanted respectively in the femoral artery of adult dog, after 2 hours, taken out and carry out sem observation.Fig. 3, Fig. 4 are respectively after the modification and the electron scanning micrograph of unmodified implanting device.Fig. 3 showed implantation after 2 hours, and the implanting device surface after the modification is almost covered by oval or flat cell, and most cells begin to sprawl at material surface.Fig. 4 showed implantation after 2 hours, platelet and fibrin that unmodified implanting device surface deposition is more.It is thus clear that the cardiovascular implantation device after the inventive method modification is capturing endothelial ancestral cell effectively, the induced material surface is endothelialization fast, has useful anticoagulation function.
Claims (3)
1. the surface orientation fixation of C D34 antibody of a titanium or titanium alloy cardiovascular implantation device or the method for CD133 antibody, its step is following:
A, activation processing
The titanium or titanium alloy cardiovascular implantation device was soaked 1-24 hour in 40-80 ℃ 0.5-5mol/L NaOH solution, take out and to be placed in the 60-90 ℃ of deionized water insulation 1-24 hour, again at 30-90 ℃ air drying;
Perhaps the titanium or titanium alloy cardiovascular implantation device was soaked 0.5-3 hour in the orthophosphoric acid solution of 0.5-5mol/L, clean drying;
Fixing of B, Avidin:
A is gone on foot implanting device after the activation processing immersed in normal saline that Avidin concentration is 0.1-5mg/m1 or the phosphate buffer 1-12 hour, take out the back with cleaning drying;
Perhaps A is gone on foot implanting device after the activation processing behind deionized water, acetone, toluene rinsing successively; Placing the toluene that contains 1%-10%3-aminopropyl three oxygen ethylsilane to be heated to 60 ℃ refluxed 4 hours; Naturally cool to room temperature preservation 12h, take out the back and clean the final vacuum drying successively with toluene, ethanol, deionized water; Again its immersion is contained the 0.1-5mg/ml Avidin and contain in 2-(N-morphine quinoline) the ethyl sulfonic acid solution of N-hydroxy-succinamide and 0.1mg/m1 of 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide, 0.25mg/m1 of 1-10mg/m1 and react 1-4h; Clean drying;
C, biotinylated protein A's is fixing:
Implanting device after B step handled immerses in the albumin solution of 0.1%-5%, and incubated at room 5-40 minute, again its immersion is contained in normal saline or the phosphate buffer of biotinylated protein A of 0.01-5mg/m1 1-12 hour, take out the back and clean drying;
D: the orientation of antibody is fixed:
The absorption that C step is obtained the implanting device of biotinylated protein A immersed in the albumin solution of 0.1%-5% 5-40 minute; Took out in the normal saline that immerses the CD34 antibody that contains 0.01-1mg/m1 or CD133 antibody in the back or the phosphate buffer 1-12 hour; Take out the back and clean drying.
2. the surface orientation fixation of C D34 antibody of a kind of titanium or titanium alloy cardiovascular implantation device according to claim 1 or the method for CD133 antibody; It is characterized in that: the cleaning of said A in the step is that deionized water for ultrasonic is cleaned; Drying dries up for nitrogen; B, C, the cleaning of D in the step are normal saline or phosphate buffer rinsing, and drying dries up for nitrogen.
3. the surface orientation fixation of C D34 antibody of a kind of titanium or titanium alloy cardiovascular implantation device according to claim 1 or the method for CD133 antibody; It is characterized in that: the said A step is with NaOH immersion, insulation and dried implanting device, fixes with carrying out the Avidin that B goes on foot after irradiation under ultraviolet ray 1-30 minute again.
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| CN102648988A (en) * | 2011-02-28 | 2012-08-29 | 刘斌 | Preparation method of stent coated with CD34 antibody |
| CN102512713A (en) * | 2011-12-30 | 2012-06-27 | 西南交通大学 | Endothelialization system for surface of induction degradable blood vessel stent |
| CN102851656A (en) * | 2012-07-25 | 2013-01-02 | 广东工业大学 | Preparation method of self-assembly silanization of pure titanium metal surface |
| CN104338182A (en) * | 2013-08-08 | 2015-02-11 | 北京精密机电控制设备研究所 | Method for coating titanium alloy surface and coating layer formed by using same |
| CN106512083B (en) * | 2016-10-14 | 2019-09-24 | 湖北大学 | A kind of preparation method of medical titanium alloy |
| CN106581742B (en) * | 2016-11-22 | 2019-06-21 | 成都迈德克科技有限公司 | A method of NO catalytic type nano coating is constructed in titanium alloy surface |
| CN111544658B (en) * | 2020-06-24 | 2022-02-01 | 中国人民解放军陆军军医大学 | Cardiovascular implant for regulating and controlling immune response and promoting intimal regeneration and preparation method thereof |
| CN113713172B (en) * | 2021-09-08 | 2023-04-11 | 深圳清华大学研究院 | In-situ endothelialization promoting coating and preparation method thereof |
| CN115869471B (en) * | 2022-03-25 | 2024-01-26 | 成都百瑞恒通医疗科技有限公司 | Anticoagulation functional material, preparation method and application thereof |
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| US5925552A (en) * | 1996-04-25 | 1999-07-20 | Medtronic, Inc. | Method for attachment of biomolecules to medical devices surfaces |
| CN101066477A (en) * | 2007-05-17 | 2007-11-07 | 中国人民解放军第三军医大学 | Biological artificial blood vessel capable of in vivo capturing endothelial ancestral cell |
| CN101172168A (en) * | 2007-10-10 | 2008-05-07 | 大连理工大学 | Metallic blood vessel bracket coating for osamine glycan load CD133 antibody and method for preparing the same |
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| US5925552A (en) * | 1996-04-25 | 1999-07-20 | Medtronic, Inc. | Method for attachment of biomolecules to medical devices surfaces |
| CN101066477A (en) * | 2007-05-17 | 2007-11-07 | 中国人民解放军第三军医大学 | Biological artificial blood vessel capable of in vivo capturing endothelial ancestral cell |
| CN101172168A (en) * | 2007-10-10 | 2008-05-07 | 大连理工大学 | Metallic blood vessel bracket coating for osamine glycan load CD133 antibody and method for preparing the same |
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