浓缩血小板的研究进展培训课件

1、浓缩血小板的研究进展浓缩血小板的研究进展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 Platelet ultrastructure. 12浓缩血小板的研究进展血小板简介及功能 Platelet ultrastructu3浓缩血小板浓缩血小板（Platelets Concentrate）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

2、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-Rich Preparation3浓缩血小板的研究进展3浓缩血小板浓缩血小板（Platelets Concentr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；4浓缩血小板的研究进展应用人群的选择 明确有传染病的患者，如乙肝、梅毒、艾滋等；4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

3、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： Hard and Soft centrifugation 离心时间离心温度 5浓缩血小板的研究进展制备临床上常用的浓缩血小板主要通过两种方法制备5浓缩血小板的商业分离系统Commercial SystemBlood Vol(ml)Centrifugation(No.of Spins)PRP Vol (ml)Platelet ConcentrationActivator(+/-)Activator:

4、PRPLeukocytes(+/-)Cascade 9-1814-91-1.5CaCl21:10-GPS 601109.3Trombin1:10+ACP9132-3None1:10-Smart PRP220-12023-204-6Trombin1:10+PRGF9-7214-322-3CaCl21:10-Magellan30-60263-9CaCl21:10+Angel40241-18None1:10+/-Genesis CS30-6014-109CaCl21:10NSSequire50251.6Trombin Bovine1:10NSPlatelex5024-6NSBatroxobin1:1

5、0+Symphony PCS55-1101NS3-6Thrombin1:10NSJP200020215-9CaCl2 and Thrombin1:10+GLO PRP8.520.65-6CaCl2 and Thrombin1:10+KYOCERA20227-8CaCl21:10+Selphyl8120.5CaCl21:10+MyCells10114-5CaCl2 and Thrombin1:10+Dr.Shins System8.5113-4CaCl2 and Thrombin1:10+PCCSCurasan6浓缩血小板的研究进展商业分离系统Commercial SystemBlood V手工

6、分离 Flow chart describing preparation of PRP. 27浓缩血小板的研究进展手工分离 Flow chart de浓缩血小板分类 L-PRP PRP P-PRP 浓缩血小板类型 PRGF L-PRF PRF R-PRF8浓缩血小板的研究进展浓缩血小板分类8浓缩血小板的研究进展9PRP. Classical manual platelet-rich plasma (PRP) protocol using a two-step centrifugation procedure. 39浓缩血小板的研究进展9PRP. Classical manual platele

7、10PRGF Commonly described protocol for Anituas PRGF. 3PPGF: plasma poor in growth factorsPRGF: plasma rich in growth factors10浓缩血小板的研究进展10PRGF Commonly described prot浓缩血小板的研究进展培训课件12StructuralDifferencesSchematic illustration of the matrix and cell architecture of the four categories of platelet con

8、centrates. 312浓缩血小板的研究进展12StructuralDifferencesSchemaActivated versus Nonactivated浓缩血小板的激活13浓缩血小板的研究进展Activated versus Nonactivate14 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14浓缩血小板的研究进展14 Activa

9、ted浓缩血小板的激活方式应该考虑以下三个15纳秒脉冲电场（Nanosecond Pulse Electric Field， nsPEF）ActivatorRepresentative voltage and current tracing for pulse electric field condition used to activate PRP. Electrical signals were measured on a 2mm cuvette containing PRP and stimulated with pulse electric field for PRP activati

10、on. Each sample was exposed to five pulses, of approximately 500 nanoseconds pulse width, 4 kV peak voltage and more than 300 A peak current per pulse .415浓缩血小板的研究进展15纳秒脉冲电场（Nanosecond Pulse Elec16Non-activatedDiabetic mouse full-thickness wound healing .516浓缩血小板的研究进展16Non-activatedDiabetic mouse 17

11、Non-activatedEffects of platelet treatment on fibroblast cultures .517浓缩血小板的研究进展17Non-activatedEffects of platNon-activatedIn vitro three-dimensional tissue defect assay .518浓缩血小板的研究进展Non-activatedIn vitro three-d19Activated versus NonactivatedSchematic depiction of the effect of physiologic and uns

12、pecific activation of platelets on wound healing .419浓缩血小板的研究进展19Activated versus Nonactivat20在创面修复中的作用Cytokines/ChemokineIL-6IL-8PF-4MIP-1RANTESFractalkineNAP2GRO-ENA-78sCD40LSDF-1Growth FactorPlatelet Derived Growth Factor, PDGF-AB/BBTransforming Growth Factor-beta, TGF-1/2Vascular Endothelial Gro

13、wth Factor, VEGFEpidermal Growth Factor, EGFBasic Fibroblast Growth Factor, bFGFInsulin like Growth Factor (IGF 1 and 2)Connective Tissue Growth Factor (CTGF)Keratinocyte Growth Factor (KGF)Hepatocyte Growth Factor(HGF)Platelet Derived Endothelial Growth Factor (PDEGF)Platelet Derived Angiogenic Fac

14、tor(PDAF)20浓缩血小板的研究进展20在创面修复中的作用Cytokines/Chemokine生长因子在创面修复中的作用Schematic illustration of the role of GFs during the different stages of the wound healing process .621浓缩血小板的研究进展生长因子在创面修复中的作用Schematic illustr22细胞因子在创面修复中的作用This is true for bacteria like MSSA, Group A Streptococcus, and Neisseria gono

15、rrhoeae as well.7. 22浓缩血小板的研究进展22细胞因子在创面修复中的作用This is true fo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23浓缩血小板的研究进展目前存在的问题缺乏统一的浓缩血小板的制备标准23浓缩血小板的参考文献1. Yeaman MR. The role of platelets in antimicrobial host defense. Clin Infect Dis 1997 25(5): 951-968;

16、quiz 969-9702. Dhurat R Sukesh M. Principles and Methods of Preparation of Platelet-Rich Plasma: A Review and Authors Perspective. J Cutan Aesthet Surg 2014 7(4): 189-1973. Dohan Ehrenfest DM, Rasmusson L Albrektsson T. Classification of platelet concentrates: from pure platelet-rich plasma (P-PRP)

17、to leucocyte- and platelet-rich fibrin (L-PRF). Trends Biotechnol 2009 27(3): 158-1674. Scherer SS, Tobalem M, Vigato E et al. Nonactivated versus thrombin-activated platelets on wound healing and fibroblast-to-myofibroblast differentiation in vivo and in vitro. Plast Reconstr Surg 2012 129(1): 46e-54e5. Frelinger Iii AL, Torres AS, Caiafa A et al. Platelet-rich plasma stimulated by pulse electric fields: Platelet activation, procoagulant markers, growth factor release and cell proliferation. Platelets 20151-86. Everts PA, Knape JT, Weibrich G et al. Platelet-rich