Shi Linying, Li Rongjuan, Wei Shuzhen, Zhou Mou, Li Lei, Lin Fang, Li Yanhui, Guo Zixuan, Zhang Wei, Chen Mingliang, Shan Guiqiu
Department of Blood Transfusion, General Hospital of Southern Theatre Command of PLA.
Department of Chemical Defense, School of Military Preventive Medicine, Amy Medical University, Chongqing, China.
Blood Coagul Fibrinolysis. 2019 Mar;30(2):58-65. doi: 10.1097/MBC.0000000000000796.
: Freeze-drying is an effective means of storing platelets. In this study, we investigated the effects of a protective agent on freeze-dried platelet-rich plasma (FD-PRP) after a 12-week preservation period. Platelet structure was measured by transmission electron microscopy (TEM), and the expression levels of procaspase activating compound (PAC)-1 and CD62P were measured by flow cytometry. The levels of transforming growth factor-beta (TGF-β), platelet-derived growth factor (PDGF) and vascular endothelial growth factor (VEGF) were determined by ELISA. The effect of FD-PRP on cell proliferation was measured by cell counting. TEM revealed that most platelets were intact, and their internal structure was evident. The expression levels of the platelet activation marker CD62P in FD-PRP and fresh PRP were 36.83% ± 8.21 and 35.47% ± 4.11, respectively, without a significant difference (P > 0.05). The expression levels of PAC-1 in FD-PRP and fresh PRP were 3.23% ± 0.49 and 2.83% ± 0.44, respectively, without a significant difference (P > 0.05). Upon activation of FD-PRP and fresh PRP by thrombin, the levels of TGF-β, PDGF and VEGF were not significantly decreased in FD-PRP. Moreover, FD-PRP promoted cell proliferation in a manner similar to that of fresh PRP. The protective agent maintained the biological activity of FD-PRP after a 12-week preservation period.
冷冻干燥是储存血小板的一种有效方法。在本研究中，我们调查了一种保护剂在12周保存期后对冻干富血小板血浆（FD-PRP）的影响。通过透射电子显微镜（TEM）测量血小板结构，通过流式细胞术测量procaspase激活化合物（PAC）-1和CD62P的表达水平。通过酶联免疫吸附测定（ELISA）确定转化生长因子-β（TGF-β）、血小板衍生生长因子（PDGF）和血管内皮生长因子（VEGF）的水平。通过细胞计数测量FD-PRP对细胞增殖的影响。TEM显示大多数血小板完整，其内部结构清晰可见。FD-PRP和新鲜PRP中血小板活化标志物CD62P的表达水平分别为36.83%±8.21和35.47%±4.11，无显著差异（P>0.05）。FD-PRP和新鲜PRP中PAC-1的表达水平分别为3.23%±0.49和2.83%±0.44，无显著差异（P>0.05）。用凝血酶激活FD-PRP和新鲜PRP后，FD-PRP中TGF-β、PDGF和VEGF的水平没有显著降低。此外，FD-PRP促进细胞增殖的方式与新鲜PRP相似。该保护剂在12周保存期后维持了FD-PRP的生物活性。
