富血小板血浆提取方法的探讨-王悦

[e-mail]【】PRP。1、30PCCSkitCurasanTrindadePRP。ELISAPDGF、TGF-β、VEGFPRPMG63。TrindadePRPCaCl2PDGF2PCCSkitPRPCaCl2TGF-β2CurasanPRPCaCl2VEGF2。3PRPP＜0．05CaCl2CaCl2PRPP＞0．05。PRPRBCMG63。3、PRPCaCl2CaCl2PRP。【】Comparisonofthemethodsforthepreparationofplatelet-richplasmaWANGYue1ZHUZhe2LIUXinming3MAYingzhi2ZHOUYanmin1YANGXufang2．1．130021ChangchunDepartmentofImplantCenterSchoolofStomatologyJilinUniversityChina2．DepartmentofPathobiologyNor-manBethuneCollegeofMedicineJilinUniversityChangchun3．ClinicallaboratoryTheSecondClinicalHospitalofJilinUniversityChangchun【Abstract】ObjectiveTocomparetheeffectsofdifferentmethodsforthepreparationofplatelet-richplasmaPRP．MethodsVe-nousbloodwasobtainedfromahealthvolunteerand30patientsofthedepartmentofophthalmology．PRPwaspreparedbyPCCSsys-temCurasansystemandTrindademethodrespectively．ThelevelsofPDGFVEGFandTGF-βinthePRPsamplesweremeasuredusingELISA．AphasecontrastmicroscopewasusedtoobservethemorphologyofMG63cellsculturedwithPRP．ResultsPDGFlevelinthePRPpreparedbyTrindademethodandtreatedwithCaCl2washigherTGF-βlevelinPRPpreparedbyPCCSandtreatedwithCaCl2andthrombasewashighertheconcentrationofVEGFinPRPpreparedbyCurasanandtreatedbyCaCl2washigher．CaCl2in-creasedPDGFlevelinthePRPpreparedbyTrindademethodTGF-βbyPCCSandCurasanandVEGFbythethreemethodsP＜0．05butthetwoactivationmethodsshowednostatisticssignificanceP＞0．05．CentrifugedPRPdidnotaffecttheobservationofthemorphologyofculturedMG63cells．ConclusionThreemethodsareeffectiveintheprepariationofPRP．Preparationatlowtem-peratureandactivationofPRPbyCaCl2orCaCl2plusthrombinmayincreasethecytokincontent．CentrifugedPRPissuitableforcellculture．【Keywords】Platelet-richplasmaGrowthfactorPreparationmethodsR331．1+43Adoi10．3969/j．issn．1001－3733．2011．05．013、。1993Ganio1plate-let-richplasmaPRP。PRP4～5。·346·JPractStomatol2011Sep275CaCl2、αplatelet-derivedgrowthfactorPDGF、-βtransforminggrowthfactorTGF-β、vascularendothelialgrowthfactorVEGF、epithelialgrowthfactorEGF、insulin-likegrowthfac-torIGF、fibroblastgrowthfac-torFGF。PRP。PRPPRPPRP。11．1MG63CaCl2hPDGF、hTGF-βhVEGFELISARD10%PAADMEMGibco、。TDL-4AThermoO-lympusIX70CO2ThermoBIO-RAD680。1．2、、、PRPPRPMG63PRP。1．2．1、130PCCSkit23iPRP13000r/min3min45s23000r/min13min。CurasanKleinostheimPRP12400r/min10min23600r/min15min。Trindade-suedamIKTrindade311600r/min10min25000r/min5min。PRP1、2～3mm23/4platelet-poolplasmaPPP1/4PRP4－80℃。3PRP1．0～1．5×1012/L。3PRP31PRP210%CaCl23500U/ml10%CaCl25。29∶1v/vPRP10min12000r/min5min。ELISAPRPPDGF、TGF-β、VEGF。1．2．225PCCSkitPRP1220℃。2。PRP9∶1v/v500U/ml10%CaCl210min12000r/min5minPRP2。1．2．32PCCSkitPRP12500U/ml10%CaCl212000r/min5min22－80℃。2PRP。1．2．4PRPMG631000r/min5min5×104/ml。24500μl。APRP、25μlBPRP、25μl。1．3GraphPadInstat3．1t、CaCl2CaCl2PDGF、TGF-βVEGF。22．1、PRPPRPCaCl2CaCl2、1。2．1．1PDGF2000096ELISA。3CaCl2TrindadePDGFq=3．727P＜0．05。3、CaCl2、CaCl2TrindadeCaCl2CaCl2q=4．335P＜0．05CaCl2PDGFCaCl21。·446·JPractStomatol2011Sep275APRPBPRP50μl10%CPRP50μl500U/ml10%9∶11PRPAPRPwithoutactivationBPRPactivatedby10%CaCl2CPRPactivatedby10%CaCl2andthrombin500U/mlin9∶1v/vFig1ComparisionofdifferentactivemethodsforPRP13PRPPDGFng/mln=30Tab1DescriptivestatisticsofthePDGFlevelsinPRPpreparedbydifferentmethodsandactivatingagentsng/mln=30PCCSCurasanTrindade299．63295．63299．68CaCl2257．51293．74335．30①②CaCl2279．79285．69266．52①CaCl233q=3．727P＜0．05②3CaCl2CaCl2q=4．335P＜0．05。2．1．2TGF-β1000ELISACaCl23PCCSCurasanq=4．431P＜0．05PCCSkitTGF-βCurasan。CurasanTrindade。PCCSkitq=5．074P＜0．01CaCl2q=3．364P＜0．05CaCl2P＞0．052。2．1．3VEGFELISA。3、CaCl2、CaCl2q=4．254P＜0．05CaCl2VEGF。PCCSCaCl2q=5．757P＜0．01q=3．674P＜0．05CaCl2P＞0．05PCCSkitPRPVEGFCaCl23。2．223PRPTGF-βng/mln=30Tab2DescriptivestatisticsoftheTGF-βng/mllevelsinPRPpreparedbydifferentmethodsandactivatingagentsng/mln=30PCCSCurasanTrindade1472．321569．351746．26CaCl21745．24①②1742．17①1761．46CaCl21885．53③1487．241673．03①CaCl2PCCSCurasanq=4．431P＜0．05②PCCSCaCl2q=3．364P＜0．05③PCCSq=5．074P＜0．0133PRPVEGFpg/mln=30Tab3DescriptivestatisticsfortheVEGFlevelsinPRPdifferentpreparedbymethodsandactivatingagentspg/mln=30PCCSCurasanTrindade731．801153．23848．05CaCl21389．80①②1472．50①1023．60①CaCl21151．47③1362．13907．70①3CaCl2q=4．254P＜0．05②PCCSCaCl2q=5．757P＜0．01③q=3．674P＜0．0540±3℃、2A。20℃、2B。PRP2C。2．3·546·JPractStomatol2011Sep275ABCPRPDPRP2、AThebuffycoatafterfirstspinatroomtemperaturecentrifugeBThebuffycoatafterfirstspinatlowtemperatureandhighspeedcentri-fugeCPRPextractiveliquidpreparedatlowtemperatureandhighspeedcentrifugeDPRPextractiveliquidpreparedatroomtemperaturecentrifugeFig2ComparisionoftheeffectofdifferenttemperatureoncentrifugeofPRPPRPPRP3。A－80℃PRPB－80℃PRP。3PRPAThePRPextractiveliquidwithoutfreezepreservationBThePRPextractiveliquidafterpreservationat－80℃Fig3Comparisionoffreezepreservationwithnonefreezepreser-vationofPRP2．4PRPA、PRP25μl24hMG634A48hMG63PBS4B。BPRP25μl48h4C。PRPMG63。31998Marx6PRPPRP。Whitman7PRP。Yamamiya8PRPPieri9Ferreira10PRP、、。PRPPRP。Froum11PRP。Schlegel12PRP。13PRP、。PRP、、、。3ELISACaCl2CaCl2CaCl2CaCl2CaCl2CaCl2CaCl2。PRP20±2℃14PRP。PRP·646·JPractStomatol2011Sep275AMG63PRP×100BMG63PRP48hMG63×100CMG63PRP48h×1004PRPAM