An optimized protocol for Platelet Rich Plasma preparation to improve its angiogenic and regenerative properties

ETULAIN J; NEGROTTO, S; MEISS, R; MENA, HA; SCHATTNER, M

Berlin

Congreso; XXVI ISTH Congress and 63rd Annual Scientific and Standardization Committee (SSC) Meeting,; 2017

ISTH

Background. Although platelet rich plasma (PRP) is used for regenerative medicine as source of platelet-derived growth factors, its effectiveness remains controversial partially due to the absence of PRP preparation protocols based on regenerative role of platelets. Aims. We aimed to optimize PRP current protocols analyzing PRP dilution (to reverse the antiangiogenic effect of plasma factors); 4ºC preincubation (as low Tº increases platelet degranulation); and supplement with fibrin cryoprecipitate (as provisional matrix for regeneration) by evaluating angiogenic and regenerative responses. Optimization was compared with current PRP-preparation methods (37ºC, no PRP dilution or supplements). Experimental design. PRP pure or 25% diluted with saline, incubated at 37 or 4ºC 30min, with or without supplementation with cryoprecipitate from frozen plasma were activated with CaCl2 (22mM). Then, supernatants were used to induce: angiogenesis in vitro (HMEC-1 proliferation, migration and tubule formation), in vivo (chorioallantoic membrane, CAM) and skin mice regeneration of full-thickness excisional wounds (perimeter closure and histology). ANOVA, n=6-12, P