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1、BURN&PLASTIC SURGERY HOSPITAL OF PLA GENERAL HOSPITAL BURNS INSTITUTE OF PLA BURN&PLASTIC SURGERY HOSPITAL OF PLA GENERAL HOSPITAL BURNS INSTITUTE OF PLA 关于浓缩血小板的研关于浓缩血小板的研究进展究进展第1页,讲稿共25张,创作于星期二血小板简介及功能血小板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、收缩粘附、聚集、释放、收缩基本功能:止血
2、凝血、维持血管内皮的完整性止血凝血、维持血管内皮的完整性Plateletultrastructure.1第2页,讲稿共25张,创作于星期二3浓缩血小板浓缩血小板(浓缩血小板(Platelets ConcentratePlatelets Concentrate)根据其临床用途可分为两类:两类:第一类第一类: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-RichP
3、reparation第3页,讲稿共25张,创作于星期二应用人群的选择 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;第4页,讲稿共25张,创作于星期二制备临床上常用的浓缩血小板主要通过两种两种方法制备商业的血小板分离系统商业的血小板分离系统手工分离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患者之间的个体差异血液的采集血液的采集:无菌原则、抗凝剂的选择(ACD-A/CPD)、抗凝剂和血液的比例(1:10)离心次数离心次数:单次离心和二次离心转速转速/离心力离心力:Ha
4、rd and Soft centrifugation 离心时间离心时间离心温度离心温度 第5页,讲稿共25张,创作于星期二商业分离系统Commercial SystemBlood Vol(ml)Centrifugation(No.of Spins)PRP Vol(ml)Platelet ConcentrationActivator(+/-)Activator:PRPLeukocytes(+/-)Cascade 9-1814-91-1.5CaCl21:10-GPS 601109.3Trombin1:10+ACP9132-3None1:10-Smart PRP220-12023-204-6Trom
5、bin1:10+PRGF9-7214-322-3CaCl21:10-Magellan30-60263-9CaCl21:10+Angel40241-18None1:10+/-Genesis CS30-6014-109CaCl21:10NSSequire50251.6Trombin Bovine1:10NSPlatelex5024-6NSBatroxobin1:10+Symphony PCS55-1101NS3-6Thrombin1:10NSJP200020215-9CaCl2 and Thrombin1:10+GLO PRP8.520.65-6CaCl2 and Thrombin1:10+KYO
6、CERA20227-8CaCl21:10+Selphyl8120.5CaCl21:10+MyCells10114-5CaCl2 and Thrombin1:10+Dr.Shins System8.5113-4CaCl2 and Thrombin1:10+PCCSCurasan第6页,讲稿共25张,创作于星期二手工分离 FlowchartdescribingpreparationofPRP.2第7页,讲稿共25张,创作于星期二浓缩血小板分类 L-PRP PRP P-PRP 浓缩血小板类型 PRGF L-PRF PRF R-PRF第8页,讲稿共25张,创作于星期二9PRPPRP.Classical
7、manualplatelet-richplasma(PRP)protocolusingatwo-stepcentrifugationprocedure.3第9页,讲稿共25张,创作于星期二10PRGFPRGFCommonlydescribedprotocolforAnituasPRGF.3PPGF:plasmapooringrowthfactorsPRGF:plasmarichingrowthfactors第10页,讲稿共25张,创作于星期二11PRFPRFChoukrounsplatelet-richfibrin(PRF)method.3第11页,讲稿共25张,创作于星期二12Structu
8、ralDifferencesSchematicillustrationofthematrixandcellarchitectureofthefourcategoriesofplateletconcentrates.3第12页,讲稿共25张,创作于星期二Activated versus Nonactivated浓缩血小板的激活第13页,讲稿共25张,创作于星期二14 Activated浓缩血小板的激活方式应该考虑以下三个三个方面:血小板激活剂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
9、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时间影响细胞因子的释放量影响细胞因子的释放量第14页,讲稿共25张,创作于星期二15纳秒脉冲电场(纳秒脉冲电场(Nanosecond Pulse Electric Field,nsPEF)ActivatorRepresentative voltage and current tracingfor pulse electric field condition used toactivatePRP.Electrical signals weremeasuredona2mmcuvettecontainingPRPandstimu
10、latedwithpulseelectricfieldforPRPactivation.Eachsamplewasexposedto five pulses,of approximately 500nanoseconds pulse width,4 kV peakvoltageandmorethan300Apeakcurrentperpulse.4第15页,讲稿共25张,创作于星期二16Non-activatedDiabeticmousefull-thicknesswoundhealing.5第16页,讲稿共25张,创作于星期二17Non-activatedEffectsofplatelett
11、reatmentonfibroblastcultures.5第17页,讲稿共25张,创作于星期二Non-activatedInvitrothree-dimensionaltissuedefectassay.5第18页,讲稿共25张,创作于星期二19Activated versus NonactivatedSchematicdepictionoftheeffectofphysiologicandunspecificactivationofplateletsonwoundhealing.4第19页,讲稿共25张,创作于星期二20在创面修复中的作用Cytokines/ChemokineIL-6IL-
12、8PF-4MIP-1RANTESFractalkineNAP2GRO-ENA-78sCD40LSDF-1Growth FactorPlateletDerivedGrowthFactor,PDGF-AB/BBTransformingGrowthFactor-beta,TGF-1/2VascularEndothelialGrowthFactor,VEGFEpidermalGrowthFactor,EGFBasicFibroblastGrowthFactor,bFGFInsulinlikeGrowthFactor(IGF1and2)ConnectiveTissueGrowthFactor(CTGF)
13、KeratinocyteGrowthFactor(KGF)HepatocyteGrowthFactor(HGF)PlateletDerivedEndothelialGrowthFactor(PDEGF)PlateletDerivedAngiogenicFactor(PDAF)第20页,讲稿共25张,创作于星期二生长因子在创面修复中的作用Schematicillustrationofthe role of GFs duringthedifferentstagesofthewound healing process.6第21页,讲稿共25张,创作于星期二22细胞因子在创面修复中的作用Thisist
14、rueforbacterialikeMSSA,GroupAStreptococcus,and Neisseria gonorrhoeae aswell.7.第22页,讲稿共25张,创作于星期二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第23页,讲稿共25张,创作于星期二参考文献1.YeamanMR.The role of platelets in antimicrobial host defense.ClinInfectDis199725(5):951-968
15、;quiz969-9702.DhuratRSukeshM.Principles and Methods of Preparation of Platelet-Rich Plasma:A Review and Authors Perspective.JCutanAesthetSurg20147(4):189-1973.DohanEhrenfestDM,RasmussonLAlbrektssonT.Classification of platelet concentrates:from pure platelet-rich plasma(P-PRP)to leucocyte-and platele
16、t-rich fibrin(L-PRF).TrendsBiotechnol200927(3):158-1674.SchererSS,TobalemM,VigatoE et al.Nonactivated versus thrombin-activated platelets on wound healing and fibroblast-to-myofibroblast differentiation in vivo and in vitro.PlastReconstrSurg2012129(1):46e-54e5.FrelingerIiiAL,TorresAS,CaiafaA et al.P
17、latelet-rich plasma stimulated by pulse electric fields:Platelet activation,procoagulant markers,growth factor release and cell proliferation.Platelets20151-86.EvertsPA,KnapeJT,WeibrichG et al.Platelet-rich plasma and platelet gel:a review.JExtraCorporTechnol200638(2):174-1877.LiHLiB.PRP as a new approach to prevent infection:preparation and in vitro antimicrobial properties of PRP,(2013).第24页,讲稿共25张,创作于星期二2022/9/16感感谢谢大大家家观观看看第25页,讲稿共25张,创作于星期二