[生物医药论文精品a]骨形成蛋白微球温敏性凝胶复合系统的制备及释药特性研究

1骨形成蛋白微球温敏性凝胶复合系统的制备及释药特性研究A1A0A2A3A4A5A6A4A7A8A4A9A10A11A12A13A14A15A16A17A18A19A20A21A22A23A24A25A17A18A25A26A27A28A28A29A28A30A31A32A33A34A35A36A37A38A39A40A41A42A43A44A45A46A47A48A49A50A51A52A53A54A55（RHBMP2）A56A57A58A59A60A61A62A63A25A64A65A66A67A68A69A56A57A40A67A68A69A56A57A70A71A72A73A74A75A76A77A78A61A79A80A58A22A81A72A82A83A84A85A86A87A88A89A88A87A90A91A92A93A23A94A95A96A97A98A91酸乙醇酸共聚物（PLGA）为材料A41A42A99A53A56A57A25A100A101A102A103A96A41A42RHBMP2A104A22A56A57A82A97A105A106A107A98A108A109A110A54A111A107A108A112A113A98A114A115MPEGPLGAA30A116A117A118A41A42A73A74A25A85A86A119A120A96A121A122A123A124A102A103A96A125A126A58A22A127A60A82A128A129微球呈现三相释药模式，而微球凝胶剂体外释药曲线符合HIGUCHI方程。采用PLGACOOH制备的RHBMP2微球，其突释明显小于以PLGA为材料制备的微球，增加内水相与油相的体积比、在内水相中加入氯化钠均可以在一定程度上减小突释。A128A130BMP微球温敏性凝胶复合系统制备工艺，具有比RHBMP2微球剂更理想的体外释药特性。A131A132A133A131A132A133A134A134A46A47A48A49A50A51A52A53A54A55A135A98A91酸乙醇酸共聚物；A105A106A107A98A108A109A110A54A111A107A108A112A113A98A114A135A56A57A136A70A71A72A73A74A136A137A58PREPARATIONOFRECOMBINANTHUMANBONEMORPHOGENETICPROTEIN2LOADEDMICROSPHERES/THERMOSENSITIVEHYDROGELSCOMPLEXANDITSINVITRORELEASECHARACTERISTICFUXUDONG,LIUHONG，TANGREN,ZHANGMENGYOU（WUHANGENERALHOSPITALOFGUANZHOUCOMMAND,WUHAN430030,CHINA）ABSTRACTOBJECTIVETOEXAMINETHEEFFECTSOFFORMULATIONFACTORSONINVITRORELEASERATEOFRECOMBINANTHUMANBONEMORPHOGENETICPROTEIN2RHBMP2LOADEDMICROSPHERESANDCOMPAREDRUGRELEASEPROFILESOFMICROSPHERESANDMICROSPHERES/THERMOSENSITIVEHYDROGELSCOMPLEXMETHODSBLANKMICROSPHERESWEREPREPAREDWITHW/O/WEMULSIONSOLVENTEVAPORATIONMETHODRHBMP2LOADEDMICROSPHERESWASPREPAREDWITHABSORPTIONMETHODTHERMOSENSITIVEHYDROGELSWEREPREPAREDWITHMPEGPLGAENZYMELINKEDIMMUNOSORBENTASSAYWASUSEDTO1A138A139A140A141A142A143A144A145A146A147A148A149A150A151A152A153A154A154A155A156A156A157A154A158A159A157A160A161A162A163A164A165A142A166A167A168A169A170A169A171A172A173A174A175TEL/FAX02787649309EMAILFUXUDONG2005YAHOOCOMCN2DETERMINEDRUGCONTENTANDINVITRORELEASERATERESULTSTHEPROFILESOFRHBMP2MICROSPHERESWERECHARACTERIZEDBYTHREEPHASE,WHILETHEINVITRORELEASEPROFILEOFRHBMP2MICROSPHERES/THERMOSENSITIVEHYDROGELSCOMPLEXCONFORMEDTOHIGUCHIEQUATIONTHEAMOUNTOFRHBMP2RELEASEDFROMMICROSPHERESINTHEINITIALBURSTPHASEWASDECRESEDMARKEDLYWHENPLGACOOHWASUSEDTOPREPAREMICROSPHERESINSTEADOFPLGAINCREASINGTHEVOLUMERATIOOFINTERNALAQUEOUSANDADDINGNACLTOINTERNALAQUEOUSPHASECOULDREDUCEINITIALRELEASECONCLUSIONRHBMP2LOADEDMICROSPHERES/THERMOSENSITIVEHYDROGELSCOMPLEXEXERTEDADESIRABLERELEASECHARACTERISTICKEYWORDSRECOMBINANTHUMANBONEMORPHOGENETICPROTEIN2PLGAMPEGPLGATHERMOSENSITIVEHYDROGELSMICROSPHERE；INITIALBURSTRELEASE牙周病是世界卫生组织列为重点防治的、人类普遍罹患的口腔疾病之一，是成年人失牙的主要原因，其治疗的最终目标是实现牙周组织包括牙槽骨、牙骨质和牙周膜的完全功能性再生，而牙周组织再生的基础是牙周膜细胞（PDLCS），但在牙周损伤部位PDLCS的来源非常有限，以致牙周组织很难达到有效地再生和重建。骨形成蛋白（BONEMORPHOGENETICPROTEIN,BMP）是目前发现的唯一能在体内诱导骨与软骨的细胞因子，业已证实BMP能够促进PDLCG86G3698G8554G2462牙周组织的G1474G38091，但是BMP在体内G9213G5242和G18250的G1328G11004G991非常G4493G7143G1207G16886失G8975，限G2058G1114其发G4649和G5224G11004。G4559G6226G17878G4464的G14659G10301G17894G17877G13007G13491一G11464是BMP的G11752G12362G9921点之一，以生G10301G2499G19489G16311G7460G7021G2058G3803的G5506G10711G6122G13447G12871G12902G2058G2070G1328为BMP牙周G15967G4628部治疗的G17745体G2475到G11752G12362G13785的G19750G11556，但是G17837G12193G13485G14659G13007G13491体G3818G18334G14659G7366G13459G5194G993是非常G10714G5831，G15932现G1998一G4462的G12373G18334G6122G993G16280G2029的G989G11468G18334G14659G8181G534723，而G1000G5506G12902G2058G2070G11004G1122牙周G15967G4628部G13485G14659G17836G4396在G1363G11004G993G1427G12573G13582点。G12520G13785以G13870G1095G18252G1069G18267G18252G1861G13870G10301为G7460G7021G2058G3803BMPG5506G10711，G17902G17819G3800G7053和G5049G14414的G1260G2282在一G4462G12255G5242G990G2499G19489G1314G12373G18334；G4570G2058G3803的G5506G10711G2010G6967在以G13870G1069G1120G18267G2345G11014G18286/G13870G1095G18252G1069G18267G18252G1861G13870G10301（MPEGPLGA）为G7460G7021G2058G3803的G1969G14026G1025，BMP在G5506G10711G1969G14026G3809G2524G13007G13491G1025的G18334G14659G17907G5242G5483到进一G8505G6925G2904。MPEGPLGAG1867有G9213G6947性，一G4462G8999G5242的MPEGPLGAG8712G9354G9094在G4472G9213G991为G14270G11013流动的G9094态G2499以G7053G1427医生G11004G14659，在体G9213G991G2499发生G14026G1969而起到控G2058G14659G10301G18334放的G1328G11004。本文主要报道G1114G3800G7053和G2058G3803G5049G14414对BMPG5506G10711以G2462BMPG5506G10711G1969G14026G2070体G3818G18334G14659G7366G13459的影响。31材料和仪器骨形成蛋白（北京环亚泰克生G10301医学技术有限公司）；G13870（G1095G18252G1069G18267G18252）G1861G13870G1030150/50（PLGA，G1095G18252G1069G18267G18252比例为5050，G2010子量为10000，山东省医疗器械G11752G12362所）；端羧基G13870G1095G18252羟基G1069G18252G1861G13870G10301（PLGACOOH,G1095G18252羟基G1069G18252比例为5050,G2010子量为10000，山东省医疗器械G11752G12362所），G13870G1069烯G18267（PVA124，日产进口G2010装，北京G2282学试G2070公司）；MPEGPLGALAGAEO的比例是821，G2010子量40005000，PEGG2010子量1000，济南岱罡生G10301技术有限公司；BMP2ELISA试G2070盒（武汉博士德试G2070公司）。数显G5347电动搅拌器（G990海司乐仪器有限公司）；JY9202D超声波细胞粉碎机（宁波新芝生G10301科技股份有限公司）；SK1快G17907混匀器（常州国华电器有限公司）；UV260型紫G3818G2010光光G5242计（日本岛津公司）；SHZ82恒G9213振荡器（常州国华电器有限公司）；BX50光学显G5506镜（日本OLYMPUS公司）；S450扫描电镜（日本HITACHI公司）；ALPHA14冷冻干燥机（CHRIST,德国）。2实验方法21BMP微球的制备空白G5506G10711采G11004W/0/WG1095G2282G9354G2070挥发法G2058G3803，取G17878量G8712加入到2ML含有200MGPLGA的G1120氯G11014烷G1025，冰浴条G1226G991超声G220G86，G9994G2530G4570其G1510入15MLG22G705的PVAG9354G9094G1025，冰浴条G1226G991G2500RPM搅拌10MING2530，G4570G17728G17907G19489G14279G2200RPM加入G225MLG8712，在G4472G9213G991G13499G13505搅拌4H，G9994G2530G17819G9400，G8939G9080，G11507空G1128G8699G2282G1120G11979干燥4G27H。BMPG5506G10711G17902G17819G2572G19480法G17745G14659，G457010MG空白G5506G10711混G5760在100L含50GML1的BMP25G705G10988G8700G18252G13543G1926G9094G1025，4G263G991G2572G1948024HG2530冷冻干燥。22BMP微球凝胶的制备G12228取MPEGPLGA4GG13634G112220MLG8712G1025，搅拌G2530G19757G13634G11224G263冰G12677G1025，G5194G7114G7114振荡，G5465MPEGPLGAG1817G2010G9354G13972G2530，G2375G548320G705MPEGPLGAG8712G1969G14026。取10MGG17745G14659G5506G10711，加入25MLG1969G14026，在冰浴条G1226G991G1817G2010混匀，G2375G5483含BMP2GML1的G5506G10711G1969G14026。23BMP微球及微球凝胶剂体外释放度的测定4G4570G17745有5G22037GBMP的G5506G10711G2010G2047混G7071在25ML的01MOLL1PH74的G11979G18252G11428G13543G1926G9094、10G705和20G705的MPEGPLGEG8712G9354G9094G1025，G12946G4506量取01MLG2010G2047G13634G11221ML的G121754G5527G12661G1025，加入1MLPH74的G11979G18252G11428G13543G1926G9094，G13634G1122恒G9213振荡器G1025，振荡G17907G5242为100RG22109MIN1、G9213G5242为（37G1021）G263，G2010G2047在05H、1、3、5、7、14、21和28D取G7691，2000RPMG12175G552710MING2530，G5335G6493G990G9177G3824，G8797G9108G10301G1025加入1MLPBS，超声15MIN，2000RPMG12175G552710MING2530，取G990G9177G90940LMLG6365110比例G12244G18334G2530，采G11004G3853G5527法G18250G13864G1825G11135G2572G19480法G8991G4462G7691G2709G1025G8543G11053的G14659G10301，G17902G17819标G1946G7366G13459计G12651BMP的含量，G16825G7053法的G7828G8991G14551G32726254000PG/ML，G8611份G7691G2709G5191G15904G8991G44622G8437。G18334放G5242为G18334放前G2530G13007G13491G1025的G14659G10301含量之G5058与G18334放前G13007G13491的G14659G10301含量的G11346G2010比。24微球粒径及表面形态观察G11004光学显G5506镜G2462扫描电镜G16278G4531G5506G10711的G3818G16278形态，G8991G4462G993G4581G1122200G1022的G12902G5464，计G12651G12651术G5191G3355G12902G5464。3结果与讨论31内水相与油相的体积比对BMP微球初始释药行为的影响以PLGA50/50为G7460G7021，G13783G4531G5415内G8712G11468与G8845G11468的体G12227比G2010G2047为15、110和120G7114G5506G10711的形态和G18334G14659G15904为。G3926G32821所G12046，G19555G11540内G8712G11468与G8845G11468的体G12227比的G3698加，G5506G10711的G12902G5464G8821有G7138显的G2476G2282（G3835G13434在20G794G220M），但是G5506G10711G15932G19766的G4392G8946G3698加5，G5506G10711在0G175HG794G22G71的G18334放G5242G19489G1314。G17837G2499能因为BMP是G17902G17819G2572G19480G1328G11004G17745G14659的，G19555G11540内G8712G11468体G12227G11468对G1955G4581，G5506G10711内部和G15932G19766的G4392G8946G1955G4581（G16277G32822），G5506G10711与BMPG9354G9094G6521G16314G19766G12227G1955G4581，BMPG15999G2572G19480的量G1955G4581，G8821有G15999G2572G19480的BMP在G18334G14659G2033G7411很快G6205G6967到G18334放G1183质，G5506G10711的G12373G18334G3698加。A176A177A176A178A176A179A176A180A176A181A182A183A184A185A186A184A187A188A189A190A191A192A193A194A195A196A197A198A199A200A201A199A202A200A203A197A198A199A200A201A199A202A200A202A204A197A198A199A200A201A199A202A200A205A204图1内水相与油相的体积比对BMP微球072H释药行为的影响5图2内水相与油相的体积比对BMP微球形态的影响A206A207A208A209A210A208A211A212A213A214A215A216A217A218A216A216A219A220A2162A21932内水相中氯化钠含量对BMP微球初始释药行为的影响G4570内G8712G11468与G8845G11468的体G12227比G3278G4462在15，G1100410G705的氯G2282G19060G9354G9094G7379G1207G8712G1328为内G8712G11468，G2499以发现G5506G10711G15932G19766的G4392G8946G7138显G3698加（G3926G3282G22所G12046），因为G19555G11540内G8712G11468G9195G17891G2399G3698G20652，G3818G11468的G8712G7368G4493G7143进入G5506G10711的内部，G15932G19766的G4392G8946G7138显G3698加（G16277G32824），和G256G22G171G257G20045G991G16278G4531到的G13479G7536G11468G1296G5506G10711在0G175HG794G22G71的G18334放G5242显G14891G19489G1314。G17837G2499能G2528G7691是因为BMP是G17902G17819G2572G19480G17745G14659，G5506G10711内部和G15932G19766的G4392G8946为BMPG9354G9094的G9195G17891提供G1114很多的G17902道，有利G1122G14659G10301的G2572G19480。A221A222A221A223A221A224A221A225A221A226A221A227A221A221A228A226A223A225A229A223A230A231A232A233A234A235A236A237A238A239A240A241A242A243A244A245A246A222A221A247A248A249A250图3内水相中氯化钠含量对BMP微球072H释药行为的影响10MA10MB10MC6图4内水相与油相的体积比对BMP微球形态的影响A251A252A253A254A255A1A0A2A3A4A14A252A253A254A255A5A12A6A24A2A3A433PLGA及PLGACOOH对BMP微球释放度的影响G3926G32825所G12046，采G11004PLGACOOHG2058G3803的G5506G10711在0G175HG794G22G71的G18334放G5242G7138显G1314G1122PLGAG5506G10711，G17837是因为BMP是一G12193碱性蛋白（G12573电点在9G170左右），G2499与PLGACOOHG2010子链G1025的羧基G2499以发生G12175子G11468互G1328G11004而G1363BMPG2572G19480量G3698加6，因此PLGACOOHG5506G10711G1025未G2572G19480的BMPG1955G4581，G12373G18334G7138显G19489G1314。A7A8A7A9A7A10A7A11A7A13A7A15A7A7A16A13A9A11A17A9A18A19A20A21A22A23A25A26A27A28A29A30A31A32A29A30A31A32A33A34A35A35A36图5PLGA种类对BMP微球072H释药行为的影响34MPEGPLGA浓度对微球凝胶剂中BMP释放度的影响G3926G32826所G12046，BMPG5506G10711G15932现G1998G989G11468G18334G14659G8181G534770G7941D的快G17907G18334放G7411。G17837是G11013G1122G15932G19766未G2572G19480G14659G10301快G17907G9354G16311所导致的，G13434有30G705左右的G14659G10301G18334放；第1DG7945D的G7114滞G7411G14659G10301G18334放G17907G5242非常G13543慢，G17837是因为G2572G19480G1122G5506G10711G15932G19766的G14659G10301已经G18334放完毕，而PLGA的G19489G16311G17907G5242较慢，G2010子量为121KDA的BMP很难从G5506G10711G1025G6205G696720MA20MB7G1998来；第5DG79414DG2530G18334G14659G17907G5242G7138显加快，是G11013G1122G19555G11540PLGA逐渐G19489G16311成G8712G9354性小G2010子，G5506G10711内部和G15932G19766的G4392隙G3698G3835，为G14659G10301的G6205G6967提供G1114G7368多G17902道。BMPG5506G10711G1969G14026G2070G8821有G15932现G1998G7138显的G989G11468G18334G14659G8181G5347，体G3818G7366G13459符G2524HIGUICHIG7053G12255，6631654215TQR09884A37其G12373G18334G7138显G1314G1122G5506G10711G2070，而在1G7945D的G18334G14659G17907G5242又略快G1122G5506G10711G2070，G2499能是混G5760在G1969G14026G1025的G5506G10711就像是包G990G1114一层衣，G20652粘G5242的G1969G14026层，G19489G1314G1114G5506G10711G990未G15999G2572G19480的BMP向G1183质G6205G6967的G17907G5242。BMPG5506G10711G1969G14026G2070在5G79414D也G8821有像G5506G10711G2070那G7691呈现快G17907G18334放的特征，G17837是因为G1969G14026层阻止G1114G8712G2010的G9195G17891，PLGAG19489G16311G17907G5242G1955慢所致。A38A39A38A40A38A41A38A42A38A43A38A380481216202428A44A45A46A47A48A49A50A51A52A53A54A55A54A55A56A57图6MPEGPLGA浓度对BMP微球释药曲线的影响4结论本文比较G1114BMPG5506G10711和BMPG5506G10711G1969G14026G2070的体G3818G18334G14659特征，G2528G7114G17836G13783G4531G1114G3800G7053和G2058G3803G5049G14414对BMPG17745G14659G5506G10711G18334放G5242的影响。G11752G12362G15932G7138BMPG5506G10711呈现G989G11468G18334G14659G8181G5347，而BMPG5506G10711G1969G14026G2070体G3818G18334G14659G7366G13459