Platelet-rich plasma lyophilization enables growth factor preservation and functionality when compared with fresh platelet-rich plasma.

AIMS: To compare levels and activity of the growth factors between fresh and lyophilized platelet-rich plasma (PRP). METHODS: Analysis of platelet concentration using fibroblast and human umbilical vein endothelial cell cultures were compared between fresh and lyophilized PRP obtained from peripheral blood. RESULTS: After lyophilization, 54% of platelets were intact whereas the fresh showed no aggregation with agonists (levels under 20%). The concentration of growth factors (VEGF, EGF, TGF-ß and PDGF) in both products were similar. Fresh and lyophilized PRPs induced proliferation in the fibroblasts at 24 h (0.303 vs 0.300, respectively). CONCLUSION: Lyophilized PRP appears to be an alternative to fresh PRP and the results evidenced the role of growth factors as a key element in the activity of this product.

In vitro study of the role of thrombin in platelet rich plasma (PRP) preparation: utility for gel formation and impact in growth factors release.

INTRODUCTION: The use of PRP has been studied for different fields, with promising results in regenerative medicine. Until now, there is no study in the literature evaluating thrombin levels in serum, used as autologous thrombin preparation. Therefore, in the present study we evaluated the role played by different thrombin concentrations in PRP and the impact in the release of growth factors. Also, different activators for PRP gel formation were evaluated. METHODS: Thrombin levels were measured in different autologous preparations: serum, L-PRP (PRP rich in leukocytes) and T-PRP (thrombin produced through PRP added calcium gluconate). L-PRP was prepared according to the literature, with platelets and leukocytes being quantified. The effect of autologous thrombin associated or not with calcium in PRP gel was determined by measuring the time of gel formation. The relationship between thrombin concentration and release of growth factors was determined by growth factors (PDGF-AA, VEGF and EGF) multiplex analysis. RESULTS: A similar concentration of thrombin was observed in serum, L-PRP and T-PRP (8.13 nM, 8.63 nM and 7.56 nM, respectively) with a high variation between individuals (CV%: 35.07, 43 and 58.42, respectively). T-PRP and serum with calcium chloride showed similar results in time to promote gel formation. The increase of thrombin concentrations (2.66, 8 and 24 nM) did not promote an increase in growth factor release. CONCLUSIONS: The technique of using serum as a thrombin source proved to be the most efficient and reproducible for promoting PRP gel formation, with some advantages when compared to other activation methods, as this technique is easier and quicker with no need of consuming part of PRP. Noteworthy, PRP activation using different thrombin concentrations did not promote a higher release of growth factors, appearing not to be necessary when PRP is used as a suspension.