Abstract
BackgroundAlternative 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.MethodsLeukocyte-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 (CaCl2) were also compared.ResultsNo 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 CaCl2 resulted in a significant increase in the PDGF-BB levels, although the HGF and IGF-1 concentrations remained unchanged.ConclusionsThe 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.
Highlights
Alternative methods of platelet-rich plasma (PRP) preparation, storage, and activation that can be stably reproduced are needed to improve PRP production
To accurately assess the outcomes of future basic and clinical studies, we considered the necessity of a PRP preparation, storage, and activation method that can be stably reproduced
A significant positive correlation was observed between hepatocyte growth factor (HGF) and platelet concentrations (r = 0.60, p = 0.002), as well as between Platelet-derived growth factor (PDGF)-BB and platelet concentrations (r = 0.83, p < 0.0001)
Summary
Alternative methods of platelet-rich plasma (PRP) preparation, storage, and activation that can be stably reproduced are needed to improve PRP production. We prepared PRP using the PRGF-Endoret Kit (BTI Biotechnology Institute, Vitoria-Gasteiz, Spain) and PRGF system IV centrifuge, and used the PRP obtained for basic and clinical research (Aoto et al, 2014; Taniguchi et al, 2018; Taniguchi et al, 2019; Yoshioka et al, 2013). Using this system, preparation is conducted with a single centrifugal separation (2100 rpm, 8 min), followed by manual separation using a dedicated pipette, which results in the isolation of leukocyte-poor, PRP (LP-PRP). To accurately assess the outcomes of future basic and clinical studies, we considered the necessity of a PRP preparation, storage, and activation method that can be stably reproduced
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