Evaluation of 24 protocols for the production of platelet-rich fibrin

BACKGROUND: The aim of this study was to evaluate 24 protocols for the production of platelet rich fibrin (PRF) produced via horizontal centrifugation to better understand cell separation following protocols at various times and speeds. METHODS: All protocols were compared utilizing a recent method to quantify cells in PRF in 1 mL sequential layers pipetted from the upper layer downwards until all 10 mL were harvested. In total, 960 complete blood counts (CBCs) were investigated. Both solid and liquid-based PRF protocols were investigated following 24 protocols involving 6 relative centrifugal force (RCF) values (100, 200, 400, 700, 1000 and 1200g) at 4 centrifugation times (3, 5, 8 and 12 min). RESULTS: In general, platelets could more easily accumulate in the upper 4 layers when compared to leukocytes owing to their lower cellular density. Protocol time seemed to have a greater impact on the final cell layer separation when compared to the effect of speed. Protocols of greater than 8 min at 400g led to no leukocyte accumulation in the upper PRF layers (found specifically within the buffy coat). Protocols at or below 200g were unable to effectively accumulate platelets/leukocytes. The optimal centrifugation speed and time for solid-PRF ranged between 400 and 700g for 8 min. It was noted that variability in patient baseline platelet/leukocyte/erythrocyte counts (hematocrit) significantly affected cell layer separation. This finding was more pronounced at lower centrifugation speeds. CONCLUSIONS: Within the investigated ranges, a protocol of 700g for 8 min presented the highest yield of platelets/leukocytes evenly distributed throughout the upper PRF layers.