Anitua Eduardo, Prado Roberto, Troya María, Zalduendo Mar, de la Fuente María, Pino Ander, Muruzabal Francisco, Orive Gorka
a BTI-Biotechnology Institute , Vitoria , Spain.
b Eduardo Anitua Foundation for Biomedical Research , Vitoria , Spain.
Platelets. 2016 Jul;27(5):459-66. doi: 10.3109/09537104.2016.1143921. Epub 2016 Mar 4.
Plasma rich in growth factors (PRGF) is a biological therapy that uses patient's own growth factors for promoting tissue regeneration. Given the current European regulatory framework in which anticoagulant solution in blood extraction tubes could be considered as a medicinal product, a new PRGF protocol has been developed. The actual protocol (PRGF-A) and the new one (PRGF-B) have been performed and compared under Good Laboratory Practices. PRGF-A protocol uses extraction tubes with 0.9 mL of trisodium citrate as anticoagulant and 50 μL of calcium chloride/mL PRGF to activate it. The PRGF-B reduces the amount of sodium citrate and calcium chloride to 0.4 mL and to 20 μL, respectively. Basic hematological parameters, platelet function, the scaffold obtaining process, growth factors content, and the biological effect were compared between both PRGF obtaining protocols.
PRGF-B protocol led to a statistically significant higher enrichment and recovery of platelets regarding to the PRGF-A. Hypotonic stress response by platelets was significantly better in the new protocol. A statistically significant decrease in the basal platelet activation status of PRGF-B compared to PRGF-A was also observed. The duration of the lag phase in the platelet aggregation assay was statistically lower for the PRGF-B protocol. Both the clotting and the clot retraction time were significantly reduced in the B protocol. A higher growth factor concentration was detected in the plasma obtained using the PRGF-B protocol. The new PRGF obtaining protocol, with a reduction in the amount of anticoagulant and activator, has even improved the actual one.
富含生长因子的血浆(PRGF)是一种生物疗法,利用患者自身的生长因子促进组织再生。鉴于当前欧洲监管框架中血液采集管中的抗凝溶液可被视为药品,已开发出一种新的PRGF方案。实际方案(PRGF-A)和新方案(PRGF-B)已在良好实验室规范下进行并比较。PRGF-A方案使用含有0.9 mL柠檬酸钠作为抗凝剂的采集管,并加入50 μL氯化钙/mL PRGF以激活它。PRGF-B方案分别将柠檬酸钠和氯化钙的量减少至0.4 mL和20 μL。比较了两种PRGF获取方案之间的基本血液学参数、血小板功能、支架获取过程、生长因子含量和生物学效应。
与PRGF-A相比,PRGF-B方案导致血小板的富集和回收率在统计学上显著更高。新方案中血小板的低渗应激反应明显更好。与PRGF-A相比,还观察到PRGF-B的基础血小板活化状态在统计学上显著降低。PRGF-B方案在血小板聚集试验中的滞后相持续时间在统计学上更低。B方案中的凝血和凝块回缩时间均显著缩短。使用PRGF-B方案获得的血浆中检测到更高的生长因子浓度。新的PRGF获取方案减少了抗凝剂和激活剂的用量,甚至改进了实际方案。