CN104548197A - Dexamethasone controllable sustained release PLGA microsphere and preparation - Google Patents
Dexamethasone controllable sustained release PLGA microsphere and preparation Download PDFInfo
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- CN104548197A CN104548197A CN201410762386.XA CN201410762386A CN104548197A CN 104548197 A CN104548197 A CN 104548197A CN 201410762386 A CN201410762386 A CN 201410762386A CN 104548197 A CN104548197 A CN 104548197A
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Abstract
The invention provides a controllable sustained release polylactic acid-glycolic acid copolymer microsphere for embedding dexamethasone. The controllable sustained-release polylactic acid-glycolic acid copolymer microsphere for embedding dexamethasone is prepared in the following method: dissolving PLGA and dexamethasone in dichloromethane, adding the mixed solution into an aqueous solution of polyvinyl alcohol, stirring, centrifuging, washing the product with distilled water and freeze drying. According to the invention, by means of the excellent dissolving capacity of dichloromethane and the emulsification of the polyvinyl alcohol, the PLGA powder and DEX solution are mixed and are prepared into the DEX-embedded PLGA microsphere. According to the invention, the surface features of the microsphere are observed and the particle size of the microsphere is measured to obtain the diameter and the surface features of the biological scaffold material, and the diameter of the PLGA blank microsphere satisfies an injected size. The sustained release effect of the biological scaffold material can be detected by DEX local release. Seen from CCK-8 measurement and lactic dehydrogenase (LDH) measurement, the microsphere has no toxicity on attached mesenchymal stem cells and generates no influence on the growth activity, so that the microsphere can be used as a biological scaffold carrier.
Description
Technical field
The invention belongs to bioengineered tissue cell scaffold material, relate to a kind of dexamethasone (DEX) controllable sustained-release Poly(D,L-lactide-co-glycolide (PLGA) microsphere and preparation method thereof.
Background technology
Existing many seminar utilize nucleus pulposus cell or mescenchymal stem cell compound cells support for the research of disc tissue engineering.In the assessment of biologic bracket material, main solid-state and preformed cytoskeleton is used to comprise network and fiber mesh.But these cytoskeletoies, during carrier for transplanted cells, need open otch to be implanted in intervertebral disc, destroy the natural enclosed mini-environment of intervertebral disc, therefore defect are obvious.This type of cytoskeleton many employings agar simultaneously, hyaluronic acid etc. are raw material, though harmless to biology, because volume is comparatively large, the original microbial environment of destroyed intervertebral disc, biological tissue's compatibility is not good, and cannot guarantee upon completion of the treatment by organism degrades.Much research has been found that adding the anti-inflammatory factors such as dexamethasone (DEX) when cell shifts can reduce the adjoint inflammation of transplanting to greatest extent, and promotes cell differentiation.Therefore, under confined environments, the sustained-release administration of antiphlogistic corticoid controls to be necessary.And previously network and fiber mesh cell scaffold material cannot realize this requirement.Therefore provide the development of novel DEX controllable sustained-release PLGA microsphere to be quite necessary for the engineering research of intervertebral disc micro-wound tissue and treatment.
Summary of the invention
The object of invention is to provide a kind of dexamethasone (DEX) controllable sustained-release Poly(D,L-lactide-co-glycolide (PLGA) microsphere (DEX/PLGA microsphere), by water-oil-water (w/o/w) emulsion technique preparation, realizes especially by following steps:
Get PLGA and DEX and be dissolved in dichloromethane, mixed solution is injected containing 2% (w/v) polyvinyl alcohol (aqueous solution of poly (vinyl alcohol, PVA), stir, slough dichloromethane, collected after centrifugation microsphere, with lyophilizing after distilled water wash and get final product.Wherein dexamethasone (DEX) and Poly(D,L-lactide-co-glycolide (PLGA) mass ratio are 1 ︰ 0.01.The consumption of dichloromethane is 0.5 times of DEX.Polyvinyl alcohol consumption is 10 times of dichloromethane.
Another object of the present invention is to provide described microsphere and is preparing the application in biological support carrier.
Preparation method of the present invention also can by the carrier of cell attachment in organism in order to obtain being injected into based on following principle design (1); (2) in order to obtain biological tissue's compatibility well to the stent model that body is nontoxic; (3) cytoskeleton is realized after a certain time by the possibility of organism degrades; (4) in order to solve the Controlled release of load medicine.
The present invention utilizes the superior solubility ability of dichloromethane, the emulsification of polyvinyl alcohol, PLGA powder and DEX solution is mixed and makes the PLGA microsphere being prepared into embedding DEX.The present invention is a kind of biomaterial emulsifying transformation method.The present invention is by observe the sign of this microsphere and particle size determination learns the diameter of this biologic bracket material and surperficial sign, and this PLGA blank microsphere diameter meets and can be injected size.By the slow release effect of DEX local release Bracket for Inspection material.Measured by CCK-8 mensuration and lactic acid dehydrogenase (LDH) and learn, the mescenchymal stem cell of this microsphere on attachment does not have toxicity and does not affect growth activity.
Accompanying drawing explanation
Fig. 1 is DEX-PLGA microsphere SEM photo under different amplification.
Fig. 2 is the DEX releasing curve diagram of DEX/PLGA microsphere.
Fig. 3 is by CCK-8 pH-value determination pH: the impact of DEX/PLGA microsphere on cell proliferation.
Fig. 4 measures DEX/PLGA microsphere to apoptotic impact by LDH release.
Detailed description of the invention
Below in conjunction with drawings and Examples the present invention made and further illustrating, but the present invention is not limited to these embodiments.
embodiment 1blank PLGA microsphere is made with 6gPLGA
Get 6g PLGA, 60mg dexamethasone (DEX) is dissolved in 30ml dichloromethane, utilize glass syringe (specification, 20G) mixed solution is injected 300ml and contain 2% (w/v) polyvinyl alcohol (poly (vinyl alcohol, PVA), in aqueous solution, stir 2-3 hour (35 DEG C) with magnetic stirring apparatus with 600rpm, slough dichloromethane, to collect microsphere after 1500rpm centrifugation 2min, with lyophilizing after distilled water wash 6 times and get final product.
embodiment 2blank PLGA microsphere is made with 12gPLGA
This blank PLGA microsphere preparation process: get 12g PLGA, 120mg DEX is dissolved in 60ml dichloromethane, utilize glass syringe (specification, 20G) mixed solution is injected the aqueous solution that 600ml contains 2% (w/v) PVA, 2-3 hour (35 DEG C) are stirred with 600rpm with magnetic stirring apparatus, slough dichloromethane, to collect microsphere after 1500rpm centrifugation 2min, with lyophilizing after distilled water wash 6 times and get final product.
embodiment 3blank PLGA microsphere is made with 24gPLGA.
This blank PLGA microsphere preparation process: get 24g PLGA, 240mg DEX is dissolved in 120ml dichloromethane, utilize glass syringe (specification, 20G) mixed solution is injected the aqueous solution that 1200ml contains 2% (w/v) PVA, 2-3 hour (35 DEG C) are stirred with 600rpm with magnetic stirring apparatus, slough dichloromethane, to collect microsphere after 1500rpm centrifugation 2min, with lyophilizing after distilled water wash 6 times and get final product.
embodiment 4
This research invention has at present been employed successfully in intervertebral disc degeneration rat tails to repair degeneration intervertebral disc, test by building rat degeneration intervertebral disc model, and by independent mescenchymal stem cell, mescenchymal stem cell merges PLGA microsphere, and mescenchymal stem cell merging DEX/PLGA microsphere injections enters degeneration intervertebral disc, to study the repair of stem cell to degeneration intervertebral disc of carrying bracket.Experimental result display DEX/PLGA microsphere regular shape under each multiple of SEM, size is more consistent, has good homogeneity (see figure 1).In DEX/PLGA microsphere, DEX is released in rapid rising in first 3 days, substantially in release profiles at zero point after 3 days, and prompting DEX sustained release (see figure 2) from DEX/PLGA microsphere.After injection DEX/PLGA microsphere, in rat disc, the mescenchymal stem cell rate of increase rises, and has compared significant difference (see figure 3) with Normal group with blank PLGA group.After rat tails injection DEX/PLGA microsphere, cell LDH discharges and Normal group and blank PLGA group no significant difference, and prompting DEX/PLGA microsphere is to mescenchymal stem cell free of toxic effects (see figure 4).DEX/PLGA microsphere can reduce local inflammation reaction, does not occur relevant rejection in experimentation, without the poisoning sign of rat, and tentative confirmation this achievement in research reliability and effectiveness.
Claims (6)
1. one kind embeds the controllable sustained-release Poly(D,L-lactide-co-glycolide microsphere of dexamethasone, it is characterized in that, realized by following preparation process: get Poly(D,L-lactide-co-glycolide and dexamethasone is dissolved in dichloromethane, mixed solution is injected the aqueous solution containing polyvinyl alcohol, dichloromethane is sloughed in stirring, collected after centrifugation microsphere, with lyophilizing after distilled water wash and get final product, wherein dexamethasone and Poly(D,L-lactide-co-glycolide mass ratio are 1:0.01.
2. a kind of controllable sustained-release Poly(D,L-lactide-co-glycolide microsphere embedding dexamethasone according to claim 1, it is characterized in that, in preparation, the consumption of dichloromethane is 0.5 times of dexamethasone, and the consumption of polyvinyl alcohol is 10 times of dichloromethane.
3. a kind of controllable sustained-release Poly(D,L-lactide-co-glycolide microsphere embedding dexamethasone according to claim 1, is characterized in that, is injected by mixed solution and contains the aqueous solution that w/v is the polyvinyl alcohol of 2%.
4. a kind of controllable sustained-release Poly(D,L-lactide-co-glycolide microsphere embedding dexamethasone according to claim 1, is characterized in that, under 35 DEG C of conditions, stirs 2-3 hour, slough dichloromethane with magnetic stirring apparatus with 600rpm.
5. a kind of controllable sustained-release Poly(D,L-lactide-co-glycolide microsphere embedding dexamethasone according to claim 1, is characterized in that, to collect microsphere after 1500rpm centrifugation 2min.
6. the application in biological support carrier prepared by a kind of controllable sustained-release Poly(D,L-lactide-co-glycolide microsphere embedding dexamethasone according to claim 1.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI630923B (en) * | 2017-09-22 | 2018-08-01 | 行政院原子能委員會核能研究所 | Surface-modified microsphere composition, application of the same, and method of preparing the same |
| CN108379582A (en) * | 2018-05-30 | 2018-08-10 | 中国人民解放军总医院 | A kind of preparation method of dexamethasone magnetic microsphere |
| CN109172606A (en) * | 2018-11-30 | 2019-01-11 | 天津欣普赛尔生物医药科技有限公司 | A kind of sustained-release micro-spheres and preparation method of the body of excretion containing mescenchymal stem cell |
| CN110252218A (en) * | 2019-07-18 | 2019-09-20 | 广东省医疗器械研究所 | Protein modified polymer microballoon composite material, preparation method and application |
| CN112807273A (en) * | 2021-02-23 | 2021-05-18 | 浙江大学 | Gene editing microneedle for treating inflammatory skin diseases and application thereof |
| CN113577030A (en) * | 2021-07-30 | 2021-11-02 | 复旦大学 | Preparation method of micro-fluidic technology-based drug microcarrier for acquired deafness |
-
2014
- 2014-12-13 CN CN201410762386.XA patent/CN104548197A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| CHENG-ZHEN LIANG等: "Dual release of dexamethasone and TGF-b3 from polymeric microspheres for stem cell matrix accumulation in a rat disc degeneration model", 《ACTA BIOMATERIALIA》 * |
| 梁成振: "荷载干细胞的TGF-β3/地塞米松/ PLGA微球用于退变椎间盘的研究", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI630923B (en) * | 2017-09-22 | 2018-08-01 | 行政院原子能委員會核能研究所 | Surface-modified microsphere composition, application of the same, and method of preparing the same |
| CN108379582A (en) * | 2018-05-30 | 2018-08-10 | 中国人民解放军总医院 | A kind of preparation method of dexamethasone magnetic microsphere |
| CN108379582B (en) * | 2018-05-30 | 2020-07-17 | 中国人民解放军总医院 | Preparation method of dexamethasone magnetic microspheres |
| CN109172606A (en) * | 2018-11-30 | 2019-01-11 | 天津欣普赛尔生物医药科技有限公司 | A kind of sustained-release micro-spheres and preparation method of the body of excretion containing mescenchymal stem cell |
| CN110252218A (en) * | 2019-07-18 | 2019-09-20 | 广东省医疗器械研究所 | Protein modified polymer microballoon composite material, preparation method and application |
| CN112807273A (en) * | 2021-02-23 | 2021-05-18 | 浙江大学 | Gene editing microneedle for treating inflammatory skin diseases and application thereof |
| CN112807273B (en) * | 2021-02-23 | 2022-06-17 | 浙江大学 | Gene editing microneedle for treating inflammatory skin diseases and application thereof |
| CN113577030A (en) * | 2021-07-30 | 2021-11-02 | 复旦大学 | Preparation method of micro-fluidic technology-based drug microcarrier for acquired deafness |
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