濃縮血小板的研究進(jìn)展培訓(xùn)課件

1、濃縮血小板的研究進(jìn)展?jié)饪s血小板的研究進(jìn)展血小板簡(jiǎn)介及功能血小板是血細(xì)胞中的一種，它是從骨髓巨核細(xì)胞的細(xì)胞質(zhì)脫落的的具有生物活性的小塊胞質(zhì)，體積小，無(wú)細(xì)胞核，呈雙面微凸的圓盤(pán)狀。生理特性：粘附、聚集、釋放、收縮基本功能：止血凝血、維持血管內(nèi)皮的完整性 Platelet ultrastructure. 12濃縮血小板的研究進(jìn)展血小板簡(jiǎn)介及功能 Platelet ultrastructu3濃縮血小板濃縮血小板（Platelets Concentrate）根據(jù)其臨床用途可分為兩類(lèi)：第一類(lèi)：體內(nèi)，是外科常用的成分輸血制品，濃縮血小板一般用于血小板減少，失血癥狀明顯而又需手術(shù)者。第二類(lèi)：體外，常用于局部軟

2、組織和骨組織的修復(fù)，主要是應(yīng)用血小板內(nèi)還有豐富的細(xì)胞因子，它們?cè)趨⑴c創(chuàng)面的修復(fù)過(guò)程中發(fā)揮重要的作用。目前文獻(xiàn)中常見(jiàn)的有四種濃縮血小板類(lèi)型，也可稱(chēng)為Platelet-Rich Preparation3濃縮血小板的研究進(jìn)展3濃縮血小板濃縮血小板（Platelets Concentr應(yīng)用人群的選擇 明確有傳染病的患者，如乙肝、梅毒、艾滋等； 腫瘤患者； 服用血小板抑制劑的或血小板功能存在障礙的患者；如口服阿司匹 林、氯吡格雷等； 血小板減少的患者；4濃縮血小板的研究進(jìn)展應(yīng)用人群的選擇 明確有傳染病的患者，如乙肝、梅毒、艾滋等；4制備臨床上常用的濃縮血小板主要通過(guò)兩種方法制備商業(yè)的血小板分離系統(tǒng)手工分

3、離制備過(guò)程中的影響因素眾多，主要包括以下幾方面：患者之間的個(gè)體差異血液的采集：無(wú)菌原則、抗凝劑的選擇（ACD-A/CPD）、抗凝劑和血液的比例（1:10）離心次數(shù)： 單次離心和二次離心轉(zhuǎn)速/離心力： Hard and Soft centrifugation 離心時(shí)間離心溫度 5濃縮血小板的研究進(jìn)展制備臨床上常用的濃縮血小板主要通過(guò)兩種方法制備5濃縮血小板的商業(yè)分離系統(tǒng)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濃縮血小板的研究進(jìn)展商業(yè)分離系統(tǒng)Commercial SystemBlood V手工

6、分離 Flow chart describing preparation of PRP. 27濃縮血小板的研究進(jìn)展手工分離 Flow chart de濃縮血小板分類(lèi) L-PRP PRP P-PRP 濃縮血小板類(lèi)型 PRGF L-PRF PRF R-PRF8濃縮血小板的研究進(jìn)展?jié)饪s血小板分類(lèi)8濃縮血小板的研究進(jìn)展9PRP. Classical manual platelet-rich plasma (PRP) protocol using a two-step centrifugation procedure. 39濃縮血小板的研究進(jìn)展9PRP. Classical manual platele

7、10PRGF Commonly described protocol for Anituas PRGF. 3PPGF: plasma poor in growth factorsPRGF: plasma rich in growth factors10濃縮血小板的研究進(jìn)展10PRGF Commonly described prot濃縮血小板的研究進(jìn)展培訓(xùn)課件12StructuralDifferencesSchematic illustration of the matrix and cell architecture of the four categories of platelet con

8、centrates. 312濃縮血小板的研究進(jìn)展12StructuralDifferencesSchemaActivated versus Nonactivated濃縮血小板的激活13濃縮血小板的研究進(jìn)展Activated versus Nonactivate14 Activated濃縮血小板的激活方式應(yīng)該考慮以下三個(gè)方面：血小板激活劑的種類(lèi) 牛凝血酶、人凝血酶、鈣劑、凝血酶受體激活肽（TRAP）、膠原、類(lèi)凝血酶-巴曲酶、反復(fù)凍融、殼聚糖、納秒脈沖電場(chǎng)（nsPEF） ；激活劑的濃度的選擇激活劑和濃縮血小板的比例影響細(xì)胞因子的釋放時(shí)間影響細(xì)胞因子的釋放量14濃縮血小板的研究進(jìn)展14 Activa

9、ted濃縮血小板的激活方式應(yīng)該考慮以下三個(gè)15納秒脈沖電場(chǎng)（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濃縮血小板的研究進(jìn)展15納秒脈沖電場(chǎng)（Nanosecond Pulse Elec16Non-activatedDiabetic mouse full-thickness wound healing .516濃縮血小板的研究進(jìn)展16Non-activatedDiabetic mouse 17

11、Non-activatedEffects of platelet treatment on fibroblast cultures .517濃縮血小板的研究進(jìn)展17Non-activatedEffects of platNon-activatedIn vitro three-dimensional tissue defect assay .518濃縮血小板的研究進(jìn)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濃縮血小板的研究進(jìn)展19Activated versus Nonactivat20在創(chuàng)面修復(fù)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濃縮血小板的研究進(jìn)展20在創(chuàng)面修復(fù)中的作用Cytokines/Chemokine生長(zhǎng)因子在創(chuàng)面修復(fù)中的作用Schematic illustration of the role of GFs during the different stages of the wound healing process .621濃縮血小板的研究進(jìn)展生長(zhǎng)因子在創(chuàng)面修復(fù)中的作用Schematic illustr22細(xì)胞因子在創(chuàng)面修復(fù)中的作用This is true for bacteria like MSSA, Group A Streptococcus, and Neisseria gono

15、rrhoeae as well.7. 22濃縮血小板的研究進(jìn)展22細(xì)胞因子在創(chuàng)面修復(fù)中的作用This is true fo目前存在的問(wèn)題缺乏統(tǒng)一的濃縮血小板的制備標(biāo)準(zhǔn)臨床應(yīng)用時(shí)能否激活仍存在爭(zhēng)議，若激活，激活劑種類(lèi)、濃度及其與濃縮血小板的比例如何確定無(wú)法確定有臨床療效的濃縮血小板中最適的血小板的濃度23濃縮血小板的研究進(jìn)展目前存在的問(wèn)題缺乏統(tǒng)一的濃縮血小板的制備標(biāo)準(zhǔn)23濃縮血小板的參考文獻(xiàn)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