浓缩血小板研究进展
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14
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, of approximately 500 nanoseconds pulse width, 4 kV peak voltage and more than 300 A peak current per pulse .[4]
Centrifugation(No.of Commercial System Blood Vol(ml)
Spins)
Cascade ® GPS Ⅲ
9-18
1
60
1
ACP®
9
1
Smart PRP2®
20-120
2
PRGF®
9-72
1
Magellan®
30-60
2
Angel®
40
2
Genesis CS®
12
浓缩血小板的激活
Activated versus Nonactivated
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 ① 影响细胞因子的释放时间 ② 影响细胞因子的释放量
1:10
-
1:10
+
1:10
-
1:10
+
1:10
-
1:10
+
1:10
+/-
1:10
NS
1.6×
Trombin Bovine
1:10
NS
NS
Batroxobin
1:10
+
3-6×
Thrombin
1:10
NS
5-9×
CaCl2 and Thrombin
1:10
+
百度文库
5-6×
CaCl2 and Thrombin
Commonly described protocol for Anitua’s PRGF. [3]
10
PRF
Choukroun’s platelet-rich fibrin (PRF) method . [3]
11
Structural Differences
Schematic illustration of the matrix and cell architecture of the four categories of platelet concentrates. [3]
浓缩血小板研究进展
Platelet ultrastructure. [1]
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
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。
[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
[3]. Dohan Ehrenfest DM, Rasmusson L Albrektsson T. Classification of platelet concentrates: from pure plateletrich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF). Trends Biotechnol 2009 27(3): 158-167
[5]. 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 20151-8
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
3
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；
30-60
1
Sequire®
50
2
Platelex®
50
2
Symphony ⅡPCS®
55-110
1
JP2000®
20
2
GLO PRP®
8.5
2
KYOCERA®
20
2
Selphyl®
8
1
MyCells®
10
1
Dr.Shin’s System®
8.5
1
PCCS®
PRP Vol (ml)
4-9 10 3 3-20 4-32 6 4 4-10 5 4-6 NS
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
[1]. Yeaman MR. The role of platelets in antimicrobial host defense. Clin Infect Dis 1997 25(5): 951-968; quiz 969970
[2]. Dhurat R Sukesh M. Principles and Methods of Preparation of Platelet-Rich Plasma: A Review and Author's Perspective. J Cutan Aesthet Surg 2014 7(4): 189-197
1:10
+
7-8× 0.5×
CaCl2 CaCl2
1:10
+
1:10
+
4-5×
CaCl2 and Thrombin
1:10
+
3-4×
CaCl2 and Thrombin
1:10
+
Flow chart describing preparation of PRP. [2]
L-PRP PRP
P-PRP
临床上常用的浓缩血小板主要通过两种方法制备
①商业的血小板分离系统 ②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：
①患者之间的个体差异 ②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）
③离心次数： 单次离心和二次离心 ④转速/离心力： Hard and Soft centrifugation ⑤离心时间 ⑥离心温度
19
在创面修复中的作用
Growth Factor
Platelet Derived Growth Factor, PDGF-AB/BB Transforming Growth Factor-beta, TGF-β1/2 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) Platelet Derived Endothelial Growth Factor (PDEGF) Platelet Derived Angiogenic Factor(PDAF)
17
In vitro three-dimensional tissue defect assay .[5]
Activated versus Nonactivated
Schematic depiction of the effect of physiologic and unspecific activation of platelets on wound healing .[4]
纳秒脉冲电场（Nanosecond Pulse Electric Field， nsPEF）
15
Non-activated
Diabetic mouse full-thickness wound healing .[5]
16
Non-activated
Effects of platelet treatment on fibroblast cultures .[5]
The End
Thanks For Your Attention
谢谢观赏
1
0.6 2 2 1
1
Platelet Concentration
Activator(+/-)
Activator:PRP Leukocytes(+/-)
1-1.5× 9.3× 2-3× 4-6× 2-3× 3-9× 1-18× 9×
CaCl2 Trombin
None Trombin
CaCl2 CaCl2 None CaCl2
Cytokines/Chemokine
IL-6 IL-8 PF-4 MIP-1α RANTES Fractalkine NAP2 GRO-α ENA-78 sCD40L SDF-1α
20
Schematic illustration of the role of GFs during the different stages of the wound healing process .[6]
浓缩血小板类型
PRGF
L-PRF PRF
R-PRF
PRP
. Classical manual platelet-rich plasma (PRP) protocol using a two-step centrifugation procedure. [3]
9
PRGF
PPGF: plasma poor in growth factors PRGF: plasma rich in growth factors
[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, <http:// www. ncbi. nlm .nih.gov/pubmed/23609458> (2013).
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 gonorrhoeae as well.[7] .
22
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
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, of approximately 500 nanoseconds pulse width, 4 kV peak voltage and more than 300 A peak current per pulse .[4]
Centrifugation(No.of Commercial System Blood Vol(ml)
Spins)
Cascade ® GPS Ⅲ
9-18
1
60
1
ACP®
9
1
Smart PRP2®
20-120
2
PRGF®
9-72
1
Magellan®
30-60
2
Angel®
40
2
Genesis CS®
12
浓缩血小板的激活
Activated versus Nonactivated
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 ① 影响细胞因子的释放时间 ② 影响细胞因子的释放量
1:10
-
1:10
+
1:10
-
1:10
+
1:10
-
1:10
+
1:10
+/-
1:10
NS
1.6×
Trombin Bovine
1:10
NS
NS
Batroxobin
1:10
+
3-6×
Thrombin
1:10
NS
5-9×
CaCl2 and Thrombin
1:10
+
百度文库
5-6×
CaCl2 and Thrombin
Commonly described protocol for Anitua’s PRGF. [3]
10
PRF
Choukroun’s platelet-rich fibrin (PRF) method . [3]
11
Structural Differences
Schematic illustration of the matrix and cell architecture of the four categories of platelet concentrates. [3]
浓缩血小板研究进展
Platelet ultrastructure. [1]
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
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。
[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
[3]. Dohan Ehrenfest DM, Rasmusson L Albrektsson T. Classification of platelet concentrates: from pure plateletrich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF). Trends Biotechnol 2009 27(3): 158-167
[5]. 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 20151-8
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
3
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；
30-60
1
Sequire®
50
2
Platelex®
50
2
Symphony ⅡPCS®
55-110
1
JP2000®
20
2
GLO PRP®
8.5
2
KYOCERA®
20
2
Selphyl®
8
1
MyCells®
10
1
Dr.Shin’s System®
8.5
1
PCCS®
PRP Vol (ml)
4-9 10 3 3-20 4-32 6 4 4-10 5 4-6 NS
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
[1]. Yeaman MR. The role of platelets in antimicrobial host defense. Clin Infect Dis 1997 25(5): 951-968; quiz 969970
[2]. Dhurat R Sukesh M. Principles and Methods of Preparation of Platelet-Rich Plasma: A Review and Author's Perspective. J Cutan Aesthet Surg 2014 7(4): 189-197
1:10
+
7-8× 0.5×
CaCl2 CaCl2
1:10
+
1:10
+
4-5×
CaCl2 and Thrombin
1:10
+
3-4×
CaCl2 and Thrombin
1:10
+
Flow chart describing preparation of PRP. [2]
L-PRP PRP
P-PRP
临床上常用的浓缩血小板主要通过两种方法制备
①商业的血小板分离系统 ②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：
①患者之间的个体差异 ②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）
③离心次数： 单次离心和二次离心 ④转速/离心力： Hard and Soft centrifugation ⑤离心时间 ⑥离心温度
19
在创面修复中的作用
Growth Factor
Platelet Derived Growth Factor, PDGF-AB/BB Transforming Growth Factor-beta, TGF-β1/2 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) Platelet Derived Endothelial Growth Factor (PDEGF) Platelet Derived Angiogenic Factor(PDAF)
17
In vitro three-dimensional tissue defect assay .[5]
Activated versus Nonactivated
Schematic depiction of the effect of physiologic and unspecific activation of platelets on wound healing .[4]
纳秒脉冲电场（Nanosecond Pulse Electric Field， nsPEF）
15
Non-activated
Diabetic mouse full-thickness wound healing .[5]
16
Non-activated
Effects of platelet treatment on fibroblast cultures .[5]
The End
Thanks For Your Attention
谢谢观赏
1
0.6 2 2 1
1
Platelet Concentration
Activator(+/-)
Activator:PRP Leukocytes(+/-)
1-1.5× 9.3× 2-3× 4-6× 2-3× 3-9× 1-18× 9×
CaCl2 Trombin
None Trombin
CaCl2 CaCl2 None CaCl2
Cytokines/Chemokine
IL-6 IL-8 PF-4 MIP-1α RANTES Fractalkine NAP2 GRO-α ENA-78 sCD40L SDF-1α
20
Schematic illustration of the role of GFs during the different stages of the wound healing process .[6]
浓缩血小板类型
PRGF
L-PRF PRF
R-PRF
PRP
. Classical manual platelet-rich plasma (PRP) protocol using a two-step centrifugation procedure. [3]
9
PRGF
PPGF: plasma poor in growth factors PRGF: plasma rich in growth factors
[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, <http:// www. ncbi. nlm .nih.gov/pubmed/23609458> (2013).
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 gonorrhoeae as well.[7] .
22
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