浓缩血小板的研究进展.ppt

,解放军总医院第一附属医院烧伤整形三科暨创面修复中心,浓缩血小板的研究进展,赵帆郝岱峰,内容,血小板简介及其功能浓缩血小板及其应用人群的选择浓缩血小板的制备方法及分类浓缩血小板的激活及使用方法浓缩血小板在创面修复中的作用目前研究中存在的问题,血小板简介及功能,血小板是血细胞中的一种，它是从骨髓巨核细胞的细胞质脱落的的具有生物活性的小块胞质，体积小，无细胞核，呈双面微凸的圆盘状。生理特性：粘附、聚集、释放、收缩基本功能：止血凝血、维持血管内皮的完整性,Plateletultrastructure.1,4,浓缩血小板,浓缩血小板（PlateletsConcentrate）根据其临床用途可分为两类：第一类：体内，是外科常用的成分输血制品，浓缩血小板一般用于血小板减少，失血症状明显而又需手术者。第二类：体外，常用于局部软组织和骨组织的修复，主要是应用血小板内还有丰富的细胞因子，它们在参与创面的修复过程中发挥重要的作用。目前文献中常见的有四种浓缩血小板类型，也可称为Platelet-RichPreparation,应用人群的选择,明确有传染病的患者，如乙肝、梅毒、艾滋等；肿瘤患者；服用血小板抑制剂的或血小板功能存在障碍的患者；如口服阿司匹林、氯吡格雷等；血小板减少的患者；,制备,临床上常用的浓缩血小板主要通过两种方法制备商业的血小板分离系统手工分离制备过程中的影响因素众多，主要包括以下几方面：患者之间的个体差异血液的采集：无菌原则、抗凝剂的选择（ACD-A/CPD）、抗凝剂和血液的比例（1:10）离心次数：单次离心和二次离心转速/离心力：HardandSoftcentrifugation离心时间离心温度,商业分离系统,手工分离,FlowchartdescribingpreparationofPRP.2,浓缩血小板分类,L-PRPPRPP-PRP浓缩血小板类型PRGFL-PRFPRFR-PRF,10,PRP,.Classicalmanualplatelet-richplasma(PRP)protocolusingatwo-stepcentrifugationprocedure.3,11,PRGF,CommonlydescribedprotocolforAnituasPRGF.3,PPGF:plasmapooringrowthfactorsPRGF:plasmarichingrowthfactors,12,PRF,Choukrounsplatelet-richfibrin(PRF)method.3,13,StructuralDifferences,Schematicillustrationofthematrixandcellarchitectureofthefourcategoriesofplateletconcentrates.3,ActivatedversusNonactivated,浓缩血小板的激活,15,Activated,浓缩血小板的激活方式应该考虑以下三个方面：血小板激活剂的种类牛凝血酶、人凝血酶、钙剂、凝血酶受体激活肽（TRAP）、胶原、类凝血酶-巴曲酶、反复冻融、壳聚糖、纳秒脉冲电场（nsPEF）；激活剂的浓度的选择激活剂和浓缩血小板的比例影响细胞因子的释放时间影响细胞因子的释放量,16,纳秒脉冲电场（NanosecondPulseElectricField，nsPEF）,Activator,RepresentativevoltageandcurrenttracingforpulseelectricfieldconditionusedtoactivatePRP.Electricalsignalsweremeasuredona2mmcuvettecontainingPRPandstimulatedwithpulseelectricfieldforPRPactivation.Eachsamplewasexposedtofivepulses,ofapproximately500nanosecondspulsewidth,4kVpeakvoltageandmorethan300Apeakcurrentperpulse.4,17,Non-activated,Diabeticmousefull-thicknesswoundhealing.5,18,Non-activated,Effectsofplatelettreatmentonfibroblastcultures.5,Non-activated,Invitrothree-dimensionaltissuedefectassay.5,20,ActivatedversusNonactivated,Schematicdepictionoftheeffectofphysiologicandunspecificactivationofplateletsonwoundhealing.4,21,在创面修复中的作用,Cytokines/ChemokineIL-6IL-8PF-4MIP-1RANTESFractalkineNAP2GRO-ENA-78sCD40LSDF-1,GrowthFactorPlateletDerivedGrowthFactor,PDGF-AB/BBTransformingGrowthFactor-beta,TGF-1/2VascularEndothelialGrowthFactor,VEGFEpidermalGrowthFactor,EGFBasicFibroblastGrowthFactor,bFGFInsulinlikeGrowthFactor(IGF1and2)ConnectiveTissueGrowthFactor(CTGF)KeratinocyteGrowthFactor(KGF)HepatocyteGrowthFactor(HGF)PlateletDerivedEndothelialGrowthFactor(PDEGF)PlateletDerivedAngiogenicFactor(PDAF),生长因子在创面修复中的作用,SchematicillustrationoftheroleofGFsduringthedifferentstagesofthewoundhealingprocess.6,23,细胞因子在创面修复中的作用,ThisistrueforbacterialikeMSSA,GroupAStreptococcus,andNeisseriagonorrhoeaeaswell.7.,目前存在的问题,缺乏统一的浓缩血小板的制备标准临床应用时能否激活仍存在争议，若激活，激活剂种类、浓度及其与浓缩血小板的比例如何确定无法确定有临床疗效的浓缩血小板中最适的血小板的浓度,参考文献,1.YeamanMR.Theroleofplateletsinantimicrobialhostdefense.ClinInfectDis199725(5):951-968;quiz969-9702.DhuratRSukeshM.PrinciplesandMethodsofPreparationofPlatelet-RichPlasma:AReviewandAuthorsPerspective.JCutanAesthetSurg20147(4):189-1973.DohanEhrenfestDM,RasmussonLAlbrektssonT.Classificationofplateletconcentrates:frompureplatelet-richplasma(P-PRP)toleucocyte-andplatelet-richfibrin(L-PRF).TrendsBiotechnol200927(3):158-1674.SchererSS,TobalemM,VigatoEetal.Nonactivatedversusthrombin-activatedplateletsonwoundhealingandfibroblast-to-myofibroblastdifferentiationinvivoandinvitro.PlastReconstrSurg2012129(1):46e-54e5.FrelingerIiiAL,TorresAS,CaiafaAetal.Platelet-richplasmastimulatedbypulseelectricfields:Plateletactivation,procoagulantmarkers,growthfactorreleaseandcellproliferation.Platelets20151-86.EvertsPA,KnapeJT,WeibrichGetal.Platelet-richplasm