Activation of Equine Platelet-Rich Plasma: Comparison of Methods and Characterization of Equine Autologous Thrombin
Activation of Equine Platelet-Rich Plasma: Comparison of Methods and Characterization of Equine Autologous Thrombin
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DOI：
10.1111/j.1532-950x.2012.01016.x
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发表时间：
2012-10-01
影响因子：
1.8
通讯作者：
Tablin, Fern
中科院分区：
文献类型：
--
作者：
Textor, Jamie A.;Tablin, Fern
目的探讨并比较临床上适用的富血小板血浆（PRP）激活方法。研究设计实验。方法制备6只马的PRP。通过4种方法（自体凝血酶、牛凝血酶、氯化钙（CaCl 2）或冻融）激活PRP。通过酶联免疫吸附测定（ELISA）定量PRP释放物中所得的PDGF-BB（其中PDGF是血小板衍生生长因子）和TGF β 1（其中TGF β是转化生长因子β）水平并进行比较。生长因子的含量也进行了比较富含血小板的凝块凝血酶或氯化钙。采用Western印迹分析和血小板聚集测定法对马自体凝血酶的组成和功能进行了表征。结果CaCl 2（23 mM）激活PRP产生显著更大的PDGF释放比任何其他方法。CaCl 2、牛凝血酶和冻融激活PRP后，TGF β释放相当。自体凝血酶在引发血小板生长因子释放方面的有效性显著低于所有其他活化方法，并且在5 U/mL时诱导的血小板聚集显著低于牛凝血酶。凝块保留了大量浓度的生长因子，并且释放物与凝块中的量在激活方法之间存在差异。结论PRP激活方法在生长因子输出以及后勤考虑方面有所不同。不推荐自体凝血酶用于PRP激活。CaCl 2（23 mM）是一种有效且廉价的PRP激活方法。由CaCl 2活化产生的PRP释放物含有80%的总PDGF含量，并且易于生产，使其成为临床使用的方便产品。
Objective To investigate and compare clinically relevant Platelet-rich plasma (PRP) activation methods. Study Design Experimental. Methods PRP was prepared from 6 equine subjects. Activation of the PRP was performed by 4 methods (autologous thrombin, bovine thrombin, calcium chloride (CaCl2), or freeze-thaw). The resultant PDGF-BB (where PDGF is platelet-derived growth factor) and TGF beta 1 (where TGF beta is transforming growth factor beta) levels in PRP releasates were quantified by Enzyme-linked immunosorbent assay (ELISA) and compared. Growth factor contents were also compared between platelet-rich clots produced by thrombin or CaCl2. The composition and function of equine autologous thrombin were characterized by Western blot analysis and platelet aggregometry. Results CaCl2 (23 mM) activation of PRP yielded significantly greater PDGF release than did any other method. TGF beta release was comparable after PRP activation by CaCl2, bovine thrombin, and freeze thaw. Autologous thrombin was significantly less effective than all other activation methods in eliciting platelet growth factor release and induced significantly less platelet aggregation than bovine thrombin at 5 U/mL. Clots retained substantial concentrations of growth factor, and the amount in the releasate versus the clot differed between activation methods. Conclusions PRP activation methods differ in terms of growth factor output as well as logistical considerations. Autologous thrombin is not recommended for PRP activation. CaCl2 (23 mM) is an effective and inexpensive method of PRP activation. The PRP releasate derived from CaCl2 activation contains 80% of the total PDGF content and is easily produced, making it a convenient product for clinical use.