CN105709279B - The preparation method of PLGA-S1P nano material and PLGA-S1P nano coating bracket - Google Patents
The preparation method of PLGA-S1P nano material and PLGA-S1P nano coating bracket Download PDFInfo
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- CN105709279B CN105709279B CN201610032781.1A CN201610032781A CN105709279B CN 105709279 B CN105709279 B CN 105709279B CN 201610032781 A CN201610032781 A CN 201610032781A CN 105709279 B CN105709279 B CN 105709279B
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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
- A61L31/08—Materials for coatings
- A61L31/10—Macromolecular materials
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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
- A61L31/148—Materials at least partially resorbable by the body
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2420/00—Materials or methods for coatings medical devices
- A61L2420/02—Methods for coating medical devices
Abstract
The purpose of the present invention is to provide a kind of PLGA-S1P nano material and the preparation methods of PLGA-S1P nano coating bracket, it is characterized by: S1P is taken solvent emulsion method to carry out PLGA coating by the method using ultrasonic disperse, it is S1P:PLGA=1:10~200 (w/w) that it, which is matched, then it takes emulsifier to diffuse to the method removal organic solvent of water, freeze-dried powder is finally made using freeze-drying concentration method.PLGA-S1P nano-granule freeze-dried powder is added in solvent, drug-carried coat liquid is used as after ultrasonic dissolution；Stainless steel stent is cleaned up, drug-carried coat liquid is sprayed at by rack surface using the high-voltage electricity method of imitating/Ultrasonic spray technology repeatedly, coating layer thickness is controlled at 5~20 μm, is cleaned using deionized water/organic solvent evaporation method, up to PLGA-S1P nano coating bracket after sterilizing.Coating material using PLGA-S1P nanoparticle as bare metal stent, can play the purpose of restenosis after inhibiting stent endoprosthesis, the present invention for prevention of arterial it is narrow/interventional therapy of obliteran provides new method.
Description
Technical field
The present invention relates to medical instruments fields, especially provide a kind of PLGA-S1P nano material and PLGA-S1P nanometers of paintings
The preparation method of layer bracket.
Background technique
Arteriarctia/occlusion seriously endangered the health of people and be increasingly becoming at present so that future the mankind it is necessary
Most great one of the public health problem faced.
Stent endoprosthesis is small with patient suffering, success rate of operation is high, few intercurrent disease, post-operative recovery is fast, the hospital stays is short etc.
Advantage, it has also become peripheral arterial caused by atherosclerosis is narrow/prefered method of obliteran treatment.Although intravascular stent
Implantation has many advantages, but restenosis (In-Stent after stent endoprosthesis as a kind of emerging technology
Restenosis, ISR) become and compares stubborn problem, the medium-long term curative effect of interventional therapy is seriously affected, and limit
The extensive use of this Minimal invasive procedures is made.Therefore, how to reduce intravascular stent restenosis rate is still intervention radiation educational circles
The difficult point and hot spot of research.In recent years, people have the following to the Mechanism Study of restenosis after stent endoprosthesis: 1) intravascular
Skin cell secretion dysfunction starts reangiostenosis；2) blood vessel endothelium injury and dysfunction, subacute stent thrombosis are formed；3)
Arterial intimal injury causes acute inflammatory reaction；4) activation of vascular smooth muscle cells, hyperplasia and migration；5) vascular remodeling.And
It has been applied to the increasing that clinical bracket for eluting medicament also inhibits endothelial cell while inhibiting smooth muscle cell proliferation at present
It grows, destroys vessel endothelialisation, postpone the normal healing of blood vessel.
Sphingosine 1-phosphate abbreviation S1P is by sphingol by sphingosine kinase-1 (SPHK1) and sphingosine kinase -2
(SPHK2) phosphorylation generate, have the active lipid-metabolism object of various biological.It has stimulation cell mitogen, resistance
The only functions such as contraction of Apoptosis, the motility of adjusting cell and adjusting smooth muscle cell skeletal actin, these lifes
Managing function is realized by acting on g protein coupled receptor.
Emerging nano material poly lactide-glycolide acid (PLGA), since it is with good biocompatibility
And biodegradability, and degradation speed is controllable, and catabolite lactic acid and hydroxyacetic acid may participate in the metabolism of human body, most
End form is excreted at carbon dioxide and water, to human body without any toxic action, and will not cause blood clotting, coalescence and hair
Thin blood vessel embolism, thus have been widely used in biomedical engineering field.PLGA has been widely used in albumen and peptide at present
The production of the controlled release carrier, medical instrument of class drug and tissue engineering bracket material.
Summary of the invention
The purpose of the present invention is to provide a kind of PLGA-S1P nano material and the systems of PLGA-S1P nano coating bracket
Preparation Method, the coating material using PLGA-S1P nanoparticle as bare metal stent, so that medicament controlled-release coating branch be made
Frame is put into narrow/occlusion intra-arterial, constantly discharges S1P, increase office to stented area by the slow degradation of PLGA
Portion organizes the concentration of S1P, the endothelialization of induction rack implanting, and inhibits the hyper-proliferative of smooth muscle cell, Jin Erda
To the purpose for inhibiting restenosis after stent endoprosthesis, the present invention for prevention of arterial it is narrow/interventional therapy of obliteran provides
New method.
The present invention specifically provides a kind of preparation method of PLGA-S1P nano material, it is characterised in that: using solvent cream
S1P is carried out PLGA coating by change method, and proportion is S1P:PLGA=1:10~200 (w/w), then uses ultrasonic dispersion by gained
Nanosphere is made in PLGA-S1P, and emulsifier is then taken to diffuse to the method removal organic solvent of water, finally dense using being lyophilized
Freeze-dried powder is made in contracting method.
Wherein, the PLGA is selected from the one or more of PLGA5005, PLGA5010, PLGA7505, PLGA7510, preferably
Proportion is S1P:PLGA=1:60~120 (w/w).
The preparation method of PLGA-S1P nano material of the present invention, which is characterized in that the preparation process of nanosphere:
1., 10~200mg PLGA5005/5010/7505/7510 is dissolved in 0.6~12ml methylene chloride or tetrachloromethane
In organic solvent (preferably methylene chloride), PLGA solution is made；
2., 1mg S1P is dissolved in 1~6ml acetone, methanol, propylene glycol or glycerine (preferably acetone), S1P solution is made；
3., under 180w Probe Ultrasonic Searching, condition of ice bath, the S1P solution of 0.6~6ml is added drop-wise to 3.2~32ml PLGA
In solution, 20~60r/min of peristaltic pump drips 3~6min of ultrasound dropwise, forms lotion；
4., under the conditions of 180w Probe Ultrasonic Searching, by 6~30ml, 2~5%PVA (Mw30,000-70,000, alcoholysis degree
87%-89%) solution pours into 3. lotion that above-mentioned steps obtain, 7~15min of ultrasound under condition of ice bath；
5., under 180w Probe Ultrasonic Searching, the lotion of step 4. is added drop-wise in the PVA solution of 50~100ml 0.1~1%,
40~100r/min of peristaltic pump drips 5~10min of ultrasound；
6., 30min~2h is rotated at 30 DEG C；
7., 10000~12000r/min of revolving speed, 5~10min of time, deionized water clean three times；
8., temperature be less than or equal to -25 DEG C under the conditions of freeze-drying at pulvis.
The present invention also provides a kind of PLGA-S1P nano coating bracket prepared using the nano material, feature exists
Be equipped with PLGA-S1P nano coating in, the bracket, the coating the preparation method comprises the following steps: by PLGA-S1P nano-granule freeze-dried powder
It is added in solvent, drug-carried coat liquid is used as after ultrasonic dissolution；Stainless steel stent is cleaned up, method/super is imitated using high-voltage electricity
Drug-carried coat liquid is sprayed at rack surface by sound wave spraying technology repeatedly, and coating layer thickness control (is preferably controlled in 10 at 5~20 μm
μm or so), it is cleaned using deionized water/organic solvent evaporation method, sterilizing (optimization ethylene oxide sterilizing) is afterwards up to PLGA-
S1P nano coating bracket.
Wherein, the PLGA-S1P nano-granule freeze-dried powder is dissolved in certain density PVA, is applied after ultrasonic dissolution as load medicine
Layer liquid, preferred 5-7 μm of the rack outer surface coating layer thickness, preferred 2-3 μm of coating on inner surface thickness.
PLGA-S1P nano coating bracket of the present invention can be used as peripheral arterial bracket (such as common iliac artery, femoral artery) and
Angiocarpy bracket application.
The present invention is directed to restenosis problem after intravascular stenting, proposes poly lactide-glycolide acid
(PLGA) restenosis after nanometer Wholly-degradable bracket prevention stent endoprosthesis is made in coated S1P.The bracket can reduce medical treatment
Expense improves patient's prognosis, has huge economic benefit and social benefit.
Detailed description of the invention
Fig. 1,10-7~10-5M S1P can promote the migration of endothelial cell.
Fig. 2,10-7~10-5M S1P can promote the proliferation of endothelial cell.
Fig. 3,10-7M S1P can promote the migration of smooth muscle cell, and 10-6~10-5M S1P can inhibit moving for smooth muscle cell
It moves.
Fig. 4,10-6~10-5M S1P can inhibit the proliferation of smooth muscle cell.
Specific embodiment
Such as non-specified otherwise, percentage of the present invention is mass percent (w/w).
Embodiment 1
One, the preparation of PLGA-S1P nanoparticle:
1,100mg PLGA5005 is dissolved in 4ml methylene chloride organic solvent, PLGA dichloromethane solution is made.
2,1mg S1P is dissolved in 5ml methanol solution, S1P solution is made.
3, under the conditions of 180w Probe Ultrasonic Searching, S1P solution (peristaltic pump 20r/min) is added drop-wise to PLGA dichloromethane solution
In (ice bath), drip ultrasonic 3min, form lotion.
4, under 180W Probe Ultrasonic Searching, the PVA (30000-70000, alcoholysis degree 87%-89%, Sigma) of 10ml 2% is molten
Liquid pours into the lotion that above-mentioned steps 3 obtain, ultrasonic 7min (ice bath).
5, under 180W Probe Ultrasonic Searching, the lotion of step 4 is added drop-wise in the PVA solution of 50mL 0.1% (peristaltic pump 40r/
Min), ultrasonic 6min is dripped.
6,30min is rotated at 30 DEG C.
7, revolving speed 12000r/min, time 10min, deionized water are cleaned three times.
8, freeze-drying is stand-by at pulvis in a low temperature of subzero 25 DEG C or so.
Two, the preparation of PLGA-S1P nanoparticle coating bracket:
High-voltage electricity imitates method: PLGA-S1P nanometers of pulvis being dissolved in 40~50% PVA, spinning solution is made, will prepared
Spinning solution be packed into 20ml syringe in, then by syringe installation, be fixed in the close frame of nanometer high-pressure electrostatic.By direct current height
Voltage source is connected with syringe needle, and cylindrical receptacle is as collection device (stainless steel stent) rotate at high speed, and through leading
Line connects ground wire；Adjust the center of syringe needle alignment collection device.Collection device rotation speed is set as 5500rpm/
Min, syringe needle are set as 25cm at a distance from collection device, spinning flow rate 0.01ml/min, voltage 20KV, when electrospinning
Between 25min.Coating layer thickness control is at 10 μm or so, and deionized water is cleaned three times, and ethylene oxide sterilizing is up to PLGA-S1P nanometers
Grain coating bracket.
Gained bracket is placed in narrow/occlusion intra-arterial, is constantly released to stented area by the slow degradation of PLGA
S1P is put, increases the concentration of local organization S1P, the endothelialization of induction rack implanting, and inhibit the excessive of smooth muscle cell
Proliferation, and then achieve the purpose that restenosis after inhibition stent endoprosthesis.
Embodiment 2
One, the preparation of PLGA-S1P nanoparticle:
1,10mg PLGA7505 is dissolved in 4ml tetrachloromethane organic solvent, PLGA tetrachloromethane solution is made.
2,1mg S1P is dissolved in 5ml acetone soln, S1P acetone soln is made.
3, under the conditions of 180w Probe Ultrasonic Searching, 5mL S1P acetone soln (peristaltic pump 20r/min) is added drop-wise to 4ml PLGA
In tetrachloromethane solution (ice bath), ultrasonic 5min is dripped, forms lotion.
4, under 180W Probe Ultrasonic Searching, the PVA (30000-70000, alcoholysis degree 87%-89%, Sigma) of 20ml 5% is molten
Liquid pours into the lotion that above-mentioned steps 3 obtain, ultrasonic 10min (ice bath).
5, under 180W Probe Ultrasonic Searching, the lotion of step 4 is added drop-wise in the PVA solution of 60mL 1% (peristaltic pump 100r/
Min), ultrasonic 8min is dripped.
6,30min is rotated at 30 DEG C.
7, revolving speed 10000r/min, time 6min, deionized water are cleaned three times.
8, freeze-drying is stand-by at pulvis in a low temperature of subzero 25 DEG C or so.
Two, the preparation of PLGA-S1P nanoparticle coating bracket:
Ultrasonic spraying method:
Organic solvent that is tetrahydrofuran or cyclohexane etc. is dispersible but cannot dissolving PLGA-S1P is made mixed solution and adds
Enter in the syringe of flush coater, syringe is installed on syringe pump, adjusts flow to 0.07ml/min.Spray parameters: power
0.08W, bracket reciprocation cycle number 12 times, carrier velocity 250rpm, 26 DEG C of environment temperature, humidity 25%.Note: spraying ginseng used
Number is obtains best rack surface state, and gained coating is smooth, 5-7 μm of external surface coating thickness, coating on inner surface thickness 2-3 μ
m。
The above embodiments merely illustrate the technical concept and features of the present invention, and its object is to allow person skilled in the art
Scholar cans understand the content of the present invention and implement it accordingly, and it is not intended to limit the scope of the present invention.It is all according to the present invention
Equivalent change or modification made by Spirit Essence, should be covered by the protection scope of the present invention.
Claims (7)
1. a kind of PLGA-S1P nano coating bracket, it is characterised in that: the bracket is equipped with PLGA-S1P nano coating, described
Coating the preparation method comprises the following steps: by PLGA-S1P nano-granule freeze-dried powder be added solvent in, after ultrasonic dissolution be used as drug-carried coat liquid；It will
Stainless steel stent cleans up, and drug-carried coat liquid is sprayed at bracket repeatedly using high-voltage electricity spinning method/ultrasonic spray method
Surface, the coating layer thickness of outer surface are 5-7 μm, and coating on inner surface is volatilized with a thickness of 2-3 μm using deionized water/organic solvent
Method cleaning, up to PLGA-S1P nano coating bracket after sterilizing；
PLGA-S1P nano-granule freeze-dried powder the preparation method comprises the following steps: using solvent emulsion method by S1P carry out PLGA coating, proportion be
S1P:PLGA=1:60~120 (w/w), then PLGA-S1P is made by nanosphere using ultrasonic dispersion, then take emulsification
Agent diffuses to the method removal organic solvent of water, and freeze-dried powder finally is made using freeze-drying concentration method；
The bracket is peripheral arterial bracket or angiocarpy bracket.
2. according to PLGA-S1P nano coating bracket described in claim 1, which is characterized in that the preparation of nano-granule freeze-dried powder
Journey:
1., 60~120mg PLGA 5005/5010/7505/7510 be dissolved in 0.6~12ml methylene chloride or tetrachloromethane have
In solvent, PLGA solution is made；
2., 1mg S1P is dissolved in 1~6ml acetone, methanol, propylene glycol or glycerine, S1P solution is made；
3., under 180w Probe Ultrasonic Searching, condition of ice bath, the S1P solution of 0.6~6ml is added drop-wise to 3.2~32ml PLGA solution
In, 20~60r/min of peristaltic pump drips 3~6min of ultrasound dropwise, forms lotion；
4., under the conditions of 180w Probe Ultrasonic Searching, the PVA solution of 6~30ml 2~5% is poured into 3. lotion that above-mentioned steps obtain
In, 7~15min of ultrasound under condition of ice bath；
5., under 180w Probe Ultrasonic Searching, the lotion of step 4. is added drop-wise in the PVA solution of 50~100ml 0.1~1%, wriggle
40~100r/min is pumped, 5~10min of ultrasound is dripped；
6., 30min~2h is rotated at 30 DEG C；
7., 10000~12000r/min of revolving speed, 5~10min of time, deionized water clean three times；
8., temperature be less than or equal to -25 DEG C under the conditions of freeze-drying at pulvis.
3. according to PLGA-S1P nano coating bracket described in claim 1, it is characterised in that: the PLGA be selected from PLGA5005,
PLGA5010, PLGA 7505, PLGA 7510 it is one or more.
4. according to PLGA-S1P nano coating bracket as claimed in claim 1 or 2, it is characterised in that: PLGA is dissolved in methylene chloride
In PLGA solution is made, be further carried out PLGA coating.
5. according to PLGA-S1P nano coating bracket described in claim 2, it is characterised in that: step 4. described in PVA alcoholysis
Degree is 87%-89%.
6. according to PLGA-S1P nano coating bracket described in claim 1, it is characterised in that: the PLGA-S1P nanoparticle freeze-drying
Powder is dissolved in certain density PVA, and drug-carried coat liquid is used as after ultrasonic dissolution.
7. PLGA-S1P nano coating bracket described in a kind of claim 1 is used as and prepares peripheral arterial bracket or angiocarpy bracket material
The application of material.
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| CN108653829A (en) * | 2017-03-31 | 2018-10-16 | 上海微创医疗器械（集团）有限公司 | A kind of drug coating, the medical instrument and preparation method that drug coating is set |
| CN107550888A (en) * | 2017-10-09 | 2018-01-09 | 大连理工大学 | A kind of preparation method for the PLGA microballoons for controlling 1 phosphoric acid sphingol to be sustained |
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| US20040170685A1 (en) * | 2003-02-26 | 2004-09-02 | Medivas, Llc | Bioactive stents and methods for use thereof |
| CN102657899A (en) * | 2012-05-22 | 2012-09-12 | 东莞科威医疗器械有限公司 | Medicament coating stent capable of preventing blood vessel restenosis and preparation method thereof |
| CN103566414A (en) * | 2013-08-13 | 2014-02-12 | 重庆大学 | Nano-microcapsule coated intravascular stent of core-shell structure and preparation method of intravascular stent |
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2016
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| US20040170685A1 (en) * | 2003-02-26 | 2004-09-02 | Medivas, Llc | Bioactive stents and methods for use thereof |
| CN102657899A (en) * | 2012-05-22 | 2012-09-12 | 东莞科威医疗器械有限公司 | Medicament coating stent capable of preventing blood vessel restenosis and preparation method thereof |
| CN103566414A (en) * | 2013-08-13 | 2014-02-12 | 重庆大学 | Nano-microcapsule coated intravascular stent of core-shell structure and preparation method of intravascular stent |
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| "Sustained delivery of sphingosine-1-phosphate using poly(lactic-co-glycolic acid)-based microparticles stimulates Akt/ERK-eNOS mediated angiogenesis and vascular maturation restoring blood flow in ischemic limbs of mice";Xun Qi等;《European Journal of Pharmacology》;20100303;第634卷(第1-3期);第121页摘要、第123页第2.3.节 * |
| Xun Qi等."Sustained delivery of sphingosine-1-phosphate using poly(lactic-co-glycolic acid)-based microparticles stimulates Akt/ERK-eNOS mediated angiogenesis and vascular maturation restoring blood flow in ischemic limbs of mice".《European Journal of Pharmacology》.2010,第634卷(第1-3期),第121页摘要、第123页第2.3.节. * |
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