Summary of the invention
The objective of the invention is to overcome above-mentioned deficiency, a kind of preparation method of intravascular stent of anticoagulant property is provided, the intravascular stent of this kind method preparation, adhesion is strong between the thin layer of its surface anticoagulant and the intravascular stent, and has the good tissue compatibility and blood compatibility.
The present invention realizes that the technical scheme that its goal of the invention adopts is: a kind of preparation method of anticoagulant intravascular stent, and its practice is:
The deposition of A, plasma polymerization allylamine function film
With pulsed plasma polymerization deposition plasma polymerizable alkenyl propylamine function film on the intravascular stent surface;
B, preparation heparin sodium mixed solution
2-(the N-morphine quinoline) ethyl sulfonic acid that takes by weighing 100~900 mass parts is dissolved in the distilled water, is made into the buffer solution that concentration is 10~100mmol/l; The N-hydroxyl succinamide that adds the heparin sodium of 100~1000 mass parts, the 1-ethyl-3-of 10~50 mass parts (3-dimethylamino-propyl) carbodiimide and 5~15 mass parts again in buffer solution again, and mix homogeneously get heparin sodium mixed solution;
The deposition of C, heparin sodium
A is gone on foot the intravascular stent that deposits plasma polymerization allylamine function film, be soaked in the B heparin sodium mixed solution in step, 4~20 ℃ were reacted 12~48 hours down, after reaction finishes, and with the abundant rinsing of distilled water, drying, promptly.
Compared with prior art, the invention has the beneficial effects as follows:
The method of using plasma plated film can plate one deck on the intravascular stent surface in conjunction with firmly and contain the plasma polymerization allylamine function film that enriches functional group's (amido), the intravascular stent that will be coated with functional membrane then is soaked in the heparin sodium aqua, and the amidine functional group in the functional membrane is realized the fixing of heparin molecule as the hydroxy functional group generation chemical reaction in reaction site and the heparin molecule.Its advantage is:
One, functional membrane and blood vessel surface adopt the plasma film coating method to form, in conjunction with firm.Functional membrane carries out covalent bonds with biomacromolecule again, and with respect to static self assembly combination and silane coupled combination, its adhesion improves greatly.Thereby make that adhesion is strong between metal material surface and the biomacromolecule, difficult drop-off has secular anticoagulation function.
Two, external during in conjunction with biomacromolecule, do not use chemical coupling agent, do not use static yet, do not damage biomolecule and proteic activity fully, so its anticoagulation function excellence.
Three, modified material-allylamine that is adopted in modifying process and heparin experimental results demonstrate and have been had the favorable tissue compatibility, blood compatibility, make intravascular stent of the present invention have fine biological safety, biocompatibility.
Experiment shows, function film surface amido concentration reaches as high as 2.43% among the present invention, and intravascular stent shows by the extracorporeal blood compatibility experiment haemolysis does not almost take place that blood compatibility is good; Reduced by 84.3% to hematoblastic adhesive capacity is the highest, hematoblastic activation amount is the highest to have reduced by 71.5%, and the anticoagulation function is strong.
Table 1 is with the intravascular stent of the inventive method preparation and only deposition plasma polymerizable alkenyl propylamine function film and the intravascular stent of heparin-binding steel not, and the hematoblastic adhesive capacity on common metallic blood vessel bracket surface and activation amount (adhesion time was 120 minutes with activationary time in 45 minutes).
Table 1
As can be seen from Table 1, the surperficial platelet adhesion amount of the intravascular stent of the inventive method preparation significantly reduces, it only is the sixth of the common adherent platelet activation amount of metallic blood vessel bracket, its hematoblastic activation amount also significantly reduces, less than 1/3rd of common metallic blood vessel bracket platelet activation amount.
Table 2 is with the intravascular stent of the inventive method preparation and only deposition plasma polymerizable alkenyl propylamine function film and the intravascular stent of heparin-binding steel not, and the platelet on common metallic blood vessel bracket surface contacts the activated partial thromboplastin time after 15 minutes with platelet poor plasma.
Table 2
Table 2 shows activated partial thromboplastin time with the intravascular stent of the inventive method preparation, and (being about to a kind of phospholipid and activator is added in the blood plasma, through after hatching, the calcium ion that adds debita spissitudo, the time that its fibrin clot forms) obtained prolongation, extended to 50.1s respectively by the 35s of common metallic blood vessel bracket.Illustrate surperficial fixed heparin molecule combine with Antithrombin III success supression the activity of partial prothrombinase, also be that the system that the intravascular stent of the inventive method preparation can effectively suppress intrinsic coagulation activates.
The concrete practice of above-mentioned A step with pulsed plasma polymerization deposition plasma polymerizable alkenyl propylamine function film on the intravascular stent surface is:
A1 sputter clean: intravascular stent is used earlier acetone, ethanol ultrasonic cleaning successively; Pack into then in the vacuum chamber of 0.5~2Pa, feeding flow is the argon of 15sccm~20sccm, and radio-frequency power is 80~120W, the back bias voltage of intravascular stent is-50~-carry out 10~30 minutes sputter clean under the condition of 100V;
A2 plasma coating: after the sputter clean, again the vacuum that makes vacuum chamber is 1~2Pa, the argon that feeds flow and be 4.5~20sccm is as carrier gas, and feeding allylamine as reacting gas, to make operating pressure be 5~20Pa, be-20 in the back bias voltage of radio-frequency power 10~100W, intravascular stent~-100V, pulse duty factor are the plasma coating that carried out under 10~100% the condition 25~90 minutes, promptly plating thickness on intravascular stent is the plasma polymerization allylamine function film of 80~150nm.
Adopt the pulsed plasma method of condition like this to carry out the deposition of plasma polymerization allylamine functional membrane, it is suitable to form thickness on the intravascular stent surface, in conjunction with excellent function film coating.
The present invention is described in further detail below in conjunction with accompanying drawing and concrete embodiment.
The specific embodiment
Embodiment 1
A kind of preparation method of anticoagulant intravascular stent, its practice is:
The deposition of A, plasma polymerization allylamine function film
A1 sputter clean: use acetone, ethanol ultrasonic cleaning successively; Pack into then in the vacuum chamber of 0.5Pa, the feeding flow is the argon of 15sccm (sccm is that the gas conversion that per minute flows into is 25 ℃, 1 ml vol under the atmospheric pressure), it is 80W that radio-frequency power is set, and the back bias voltage of intravascular stent is-50V to carry out 10 minutes time sputter clean;
A2 plasma coating: after the sputter clean, again the vacuum that makes vacuum chamber is 1Pa, feeding is as the argon of carrier gas, the flow of argon is 4.5sccm, with allylamine as reacting gas, radio-frequency power be 10W, operating pressure be the back bias voltage of 5Pa, intravascular stent for-20V, pulse duty factor are 10%, 25 minutes time, promptly plating thickness on intravascular stent is the plasma polymerization allylamine function film of 80nm.
B, preparation heparin sodium mixed solution
2-(the N-morphine quinoline) ethyl sulfonic acid that takes by weighing 100 mass parts is dissolved in the distilled water, is made into the buffer solution that concentration is 10mmol/1; The N-hydroxyl succinamide that adds the heparin sodium of 100 mass parts, the 1-ethyl-3-of 10 mass parts (3-dimethylamino-propyl) carbodiimide and 5 mass parts again in buffer solution again, and mix homogeneously get heparin sodium mixed solution;
The deposition of C, heparin sodium
A is gone on foot the intravascular stent that deposits plasma polymerization allylamine function film, be soaked in the B heparin sodium mixed solution in step, 4 ℃ were reacted 48 hours down, after reaction finishes, and with the abundant rinsing of distilled water, drying, promptly.
Embodiment 2
A kind of preparation method of anticoagulant intravascular stent, its practice is:
The deposition of A, plasma polymerization allylamine function film
A1 sputter clean: use acetone, ethanol ultrasonic cleaning successively; Pack into then in the vacuum chamber of 2Pa, feeding flow is the argon of 20sccm, and it is 120W that radio-frequency power is set, and the back bias voltage of intravascular stent is-100V to carry out the sputter clean of 30 minutes time;
A2 plasma coating: after the sputter clean, again the vacuum that makes vacuum chamber is 2Pa, feeding flow is the argon as carrier gas of 20sccm, with allylamine as reacting gas, radio-frequency power be 100W, operating pressure be the back bias voltage of 20Pa, intravascular stent for-100V, pulse duty factor are 100%, 90 minutes time, promptly plating thickness on intravascular stent is the plasma polymerization allylamine function film of 150nm.
B, preparation heparin sodium mixed solution
2-(the N-morphine quinoline) ethyl sulfonic acid that takes by weighing 900 mass parts is dissolved in the distilled water, is made into the buffer solution that concentration is 100mmol/l; The N-hydroxyl succinamide that adds the heparin sodium of 1000 mass parts, the 1-ethyl-3-of 50 mass parts (3-dimethylamino-propyl) carbodiimide and 15 mass parts again in buffer solution again, and mix homogeneously get heparin sodium mixed solution;
The deposition of C, heparin sodium
A is gone on foot the intravascular stent that deposits plasma polymerization allylamine function film, be soaked in the B heparin sodium mixed solution in step, 20 ℃ were reacted 12 hours down, after reaction finishes, and with the abundant rinsing of distilled water, drying, promptly.
Embodiment 3
A kind of preparation method of anticoagulant intravascular stent, its practice is:
The deposition of A, plasma polymerization allylamine function film
A1 sputter clean: use acetone, ethanol ultrasonic cleaning successively; Pack into then in the vacuum chamber of 1Pa, feeding flow is the argon of 18sccm, and it is 100W that radio-frequency power is set, and the back bias voltage of intravascular stent is-75V to carry out the sputter clean of 20 minutes time;
A2 plasma coating: after the sputter clean, again the vacuum that makes vacuum chamber is 1.5Pa, feeding flow is the argon as carrier gas of 15sccm, with allylamine as reacting gas, radio-frequency power be 50W, operating pressure be the back bias voltage of 15Pa, intravascular stent for-60V, pulse duty factor are 50%, 45 minutes time, promptly plating thickness on intravascular stent is the plasma polymerization allylamine function film of 100nm.
B, preparation heparin sodium mixed solution
2-(the N-morphine quinoline) ethyl sulfonic acid that takes by weighing 400 mass parts is dissolved in the distilled water, is made into the buffer solution that concentration is 50mmol/l; The N-hydroxyl succinamide that adds the heparin sodium of 500 mass parts, the 1-ethyl-3-of 25 mass parts (3-dimethylamino-propyl) carbodiimide and 10 mass parts again in buffer solution again, and mix homogeneously get heparin sodium mixed solution;
The deposition of C, heparin sodium
A is gone on foot the intravascular stent that deposits plasma polymerization allylamine function film, be soaked in the B heparin sodium mixed solution in step, 12 ℃ were reacted 24 hours down, after reaction finishes, and with the abundant rinsing of distilled water, drying, promptly.
Embodiment 4
A kind of preparation method of anticoagulant intravascular stent, its practice is:
The deposition of A, plasma polymerization allylamine function film
A1 sputter clean: use acetone, ethanol ultrasonic cleaning successively; Pack into then in the vacuum chamber of 1.5Pa, feeding flow is the argon of 10sccm, and it is 60W that radio-frequency power is set, and the back bias voltage of intravascular stent is-90V to carry out the sputter clean of 20 minutes time;
A2 plasma coating: after the sputter clean, again the vacuum that makes vacuum chamber is 1Pa, feeding is as the argon (flow is 12sccm) of carrier gas, with allylamine as reacting gas, radio-frequency power be 50W, operating pressure be the back bias voltage of 12Pa, intravascular stent for-80V, pulse duty factor are 30%, 45 minutes time, promptly plating thickness on intravascular stent is the plasma polymerization allylamine function film of 90nm.
B, preparation heparin sodium mixed solution
2-(the N-morphine quinoline) ethyl sulfonic acid that takes by weighing 600 mass parts is dissolved in the distilled water, is made into the buffer solution that concentration is 60mmol/l; The N-hydroxyl succinamide that adds the heparin sodium of 400 mass parts, the 1-ethyl-3-of 25 mass parts (3-dimethylamino-propyl) carbodiimide and 12 mass parts again in buffer solution again, and mix homogeneously get heparin sodium mixed solution;
The deposition of C, heparin sodium
A is gone on foot the intravascular stent that deposits plasma polymerization allylamine function film, be soaked in the B heparin sodium mixed solution in step, 10 ℃ were reacted 28 hours down, after reaction finishes, and with the abundant rinsing of distilled water, drying, promptly.
Fig. 1 a and Fig. 1 b are respectively the stereoscan photograph that contacts 120 minutes samples of cultivation with the platelet on the intravascular stent of the inventive method preparation and common metallic blood vessel bracket surface with fresh healthy people's platelet rich plasma.
The metallic blood vessel bracket surface common as can be seen from Fig. 1 b has a large amount of platelet adhesions, and platelet is out of shape and reunion is more serious, has a large amount of pseudopodium to produce, and this shows in common metallic blood vessel bracket surface platelet activation degree bigger.
Fig. 1 a shows then and the intravascular stent of the inventive method preparation that its surface shows as platelet adhesion, activation and the distortion of low quantity.
Fig. 2 a and Fig. 2 b are respectively the surface topography that implanted 30 days with the intravascular stent of the inventive method preparation and common metallic blood vessel bracket.
From Fig. 2 a and Fig. 2 b as can be seen: after implanting 30 days, all do not produce thrombosis with the intravascular stent of the inventive method preparation and common metallic blood vessel bracket surface, blood vessel is more unobstructed.Among Fig. 2 b, comparatively serious hamartoplasia phenomenon has appearred in common most of zone, metallic blood vessel bracket surface, and outgrowth tissue is with blood vessel adhesion.And among Fig. 2 a, the intravascular stent surface texture of the inventive method preparation is extremely thin, smoothly be transparence.
Fig. 3 a and Fig. 3 b are respectively the surface topography that implanted 70 days with the intravascular stent of the inventive method preparation and common metallic blood vessel bracket.
From Fig. 3 b as can be seen, serious obstruction has taken place in the blood vessel of the common metal intravascular stent of implantation after 70 days; Fig. 3 a then as can be seen, though implant the phenomenon that hamartoplasia has also taken place on the intravascular stent surface of the inventive method preparation after 70 days, blood vessel is still comparatively unobstructed.
Fig. 4 a is for strutting the stereoscan photograph of adhering to situation of rear surface thin film with the intravascular stent of system of the present invention; Fig. 4 b and Fig. 4 c are respectively the stereoscan photograph of adhering to situation of sedimentary TiO film in metal surface and Ti film stretching rear surface thin film.
Fig. 4 b and Fig. 4 c coming off and metaboly of a large amount of thin film occurred after showing that then sedimentary TiO film in metal surface and Ti film stretch.And the intravascular stent of the present invention's preparation among Fig. 4 a, the surface is evenly smooth, does not have to come off because of strutting the deformation and the thin film that bring; Illustrate that adhesion is strong between plasma polymerization allylamine function film layer of the present invention and the intravascular stent, the pliability of thin film is good, can not be out of shape because of support struts.