Optimization of leukocyte-poor platelet-rich plasma preparation: a validation study of leukocyte-poor platelet-rich plasma obtained using different preparer, storage, and activation methods.

BACKGROUND: Alternative methods of platelet-rich plasma (PRP) preparation, storage, and activation that can be stably reproduced are needed to improve PRP production. The purpose of this study was to investigate the effect of the preparer's experience on the quality of prepared PRP, chronological changes occurring in PRP, and the effect of the activation procedures on the release of several growth factors from PRP, using PRP prepared with the PRGF-Endoret Kit. METHODS: Leukocyte-poor PRP samples from seventeen healthy volunteers were prepared using the PRGF-Endoret Kit and the PRGF IV System Centrifuge. The platelet and leukocyte concentrations were compared based on the preparer's experience. The concentrations of platelets, hepatocyte growth factor (HGF), platelet-derived growth factor-BB (PDGF-BB), and insulin-like growth factor-1 (IGF-1) were determined at 0 and 60 min after PRP preparation, and compared. Concentrations of the above growth factors from PRP activated by freeze-thaw cycling and by calcium chloride (CaCl(2)) were also compared. RESULTS: No significant difference was observed in the platelet concentrations and leukocyte contamination rates, based on the preparer's experience. At 60 min after PRP preparation, the platelet concentration decreased significantly, while the HGF, PDGF-BB, and IGF-1 concentrations remained unchanged. Activation with CaCl(2) resulted in a significant increase in the PDGF-BB levels, although the HGF and IGF-1 concentrations remained unchanged. CONCLUSIONS: The results of this study show that leukocyte-poor PRP prepared using the PRGF-Endoret Kit did not result in any qualitative difference that depended on the experience of the preparer. However, PRP preparation required standardization in terms of the time of blood count measurement. Growth factor concentrations in PRP differed according to the platelet-activation method used.