Optimization of pure platelet‑rich plasma preparation: A comparative study of pure platelet‑rich plasma obtained using different centrifugal conditions in a single‑donor model

While it has been proved that centrifugal conditions for pure platelet‑rich plasma (P‑PRP) preparation influence the cellular composition of P‑PRP obtained, the optimal centrifugal conditions to prepare P‑PRP have not yet been identified. In the present study, platelet‑containing plasma (PCP) was prepared with the first‑spin of different double‑spin methods and P‑PRP was prepared with different double‑spin methods. Whole‑blood analysis was performed to evaluate the cellular composition of PCP and P‑PRP. The basal and ADP‑induced CD62P expression rates of platelets were assessed by flow cytometry to evaluate the function of platelets in PCP and P‑PRP. Enzyme‑linked immune sorbent assay was performed to quantify interleukin‑1β, tumor necrosis factor‑α, platelet‑derived growth factor AB and transforming growth factor β1 concentrations of PCP and P‑PRP. Correlations between the cellular characteristics and cytokine concentrations of P‑PRP were analyzed by Pearson correlation analysis. Effects of P‑PRP on the proliferation, survival and migration of human bone marrow‑derived mesenchymal stem cells and human articular chondrocytes were evaluated by a Cell Counting Kit‑8 assay, live/dead staining and Transwell assay, respectively. The results showed that centrifugation at 160 x g for 10 min and 250 x g for 15 min successively captured and concentrated platelets and growth factors significantly more efficiently with preservation of platelet function compared with other conditions (P<0.05). The correlation analysis showed that the similar leukocyte concentrations and leukocyte‑reducing efficiencies resulted in similar pro‑inflammatory cytokine concentrations in P‑PRP (P>0.05) and the maximization of platelet concentration, platelet enrichment factor, platelet capture efficiency and platelet function resulted in the maximization of growth factor concentrations in P‑PRP obtained using the optimal conditions (P<0.05). Compared with P‑PRP obtained under other conditions, P‑PRP obtained under the optimal conditions significantly promoted the proliferation and migration of cells (P<0.05) and did not alter cell survival (P>0.05). Therefore, centrifugation at 160 x g for 10 min and 250 x g for 15 min successively with removal of the buffy coat as a crucial step may provide an optimal preparation system of P‑PRP for clinical application.