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1、,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,Platelet ultrastructure. 1,4,浓缩血小板,浓缩血小板（Platelets Concentrate）根
2、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-Rich Preparation,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
3、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,商业分离系统,手工分离,Flow chart describing preparation of PRP. 2,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,10,PRP,. Classical manual platelet-rich plasma (P
4、RP) protocol using a two-step centrifugation procedure. 3,11,PRGF,Commonly described protocol for Anituas PRGF. 3,PPGF: plasma poor in growth factors PRGF: plasma rich in growth factors,12,PRF,Choukrouns platelet-rich fibrin (PRF) method . 3,13,Structural Differences,Schematic illustration of the ma
5、trix and cell architecture of the four categories of platelet concentrates. 3,Activated versus Nonactivated,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（Nanosecond Pulse
6、Electric Field， nsPEF）,Activator,Representative 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 activation. Each sample was exposed to five pulses, o
7、f approximately 500 nanoseconds pulse width, 4 kV peak voltage and more than 300 A peak current per pulse .4,17,Non-activated,Diabetic mouse full-thickness wound healing .5,18,Non-activated,Effects of platelet treatment on fibroblast cultures .5,Non-activated,In vitro three-dimensional tissue defect
8、 assay .5,20,Activated versus Nonactivated,Schematic depiction of the effect of physiologic and unspecific activation of platelets on wound healing .4,21,在创面修复中的作用,Cytokines/Chemokine IL-6 IL-8 PF-4 MIP-1 RANTES Fractalkine NAP2 GRO- ENA-78 sCD40L SDF-1,Growth Factor Platelet Derived Growth Factor,
9、PDGF-AB/BB Transforming Growth Factor-beta, TGF-1/2 Vascular Endothelial Growth Factor, VEGF Epidermal Growth Factor, EGF Basic Fibroblast Growth Factor, bFGF Insulin like Growth Factor (IGF 1 and 2) Connective Tissue Growth Factor (CTGF) Keratinocyte Growth Factor (KGF) Hepatocyte Growth Factor(HGF
10、) Platelet Derived Endothelial Growth Factor (PDEGF) Platelet Derived Angiogenic Factor(PDAF),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 .6,23,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
11、 gonorrhoeae as well.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. Yeaman MR. The role of platelets in antimicrobial host defense. Clin Infect Dis 1997 25(5): 951-968; quiz 969-970 2. Dhurat R Sukesh M. Principles and Methods of Preparation
12、of Platelet-Rich Plasma: A Review and Authors Perspective. J Cutan Aesthet Surg 2014 7(4): 189-197 3. Dohan Ehrenfest DM, Rasmusson L Albrektsson T. Classification of platelet concentrates: from pure platelet-rich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF). Trends Biotechnol 2009
13、27(3): 158-167 4. 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-54e 5. Frelinger Iii AL, Torres AS, Caiafa A et al. Platelet-rich plas
14、ma stimulated by pulse electric fields: Platelet activation, procoagulant markers, growth factor release and cell proliferation. Platelets 20151-8 6. Everts PA, Knape JT, Weibrich G et al. Platelet-rich plasma and platelet gel: a review. J Extra Corpor Technol 2006 38(2): 174-187 7. Li H Li B. PRP as a new approach to prevent infection: preparation and in vitro antimicrobial properties of PRP, (2013).,The End Thanks For Your Attention,
