Effect of Centrifugation Methods on the Regenerative Potential of Concentrated Platelet-Rich Fibrin: An In Vitro Study

Objective A new type of platelet concentrate used in regenerative periodontal therapy, called concentrated platelet-rich fibrin (C-PRF), contains more growth factors than previous generations of platelet concentrate. Compared to fixed-angle centrifugation, a recently discovered technique called horizontal centrifugation can produce C-PRF with a more evenly distributed quantity of platelets and improved biological properties. This study aims to determine the effect of horizontal and fixed-angle centrifugation methods on the release of transforming growth factor β1 (TGF-β1) and platelet-derived growth factor AB (PDGF-AB), as well as on the migration and proliferation of osteoblast cells. Materials and Methods This study used two test groups: a horizontal group and a fixed-angle group. The release of growth factors TGF-β1 and PDGF-AB was measured at five time points: the first, third, seventh, 10^th, and 14th days of incubation, using the enzyme-linked immunosorbent assay. To determine the effect of C-PRF type on osteoblast cell migration, a scratch wound healing assay was conducted 24 hours after incubation. Osteoblast cell proliferation was counted using a 450-nm microplate reader to determine the number of cells in the well plate on the first, third, and fifth days, with the Cell Counting Kit-8. The data were examined using two-way ANOVA (analysis of variance) and post hoc LSD (least significant difference). Results The findings indicated a statistically significant increase in the release of TGF-β1 and PDGF-AB in the horizontal group compared to the fixed-angle group over a 14-day period. The horizontal group's TGF-β1 release increased by a factor of 1.5 compared to the fixed-angle group, peaking on day 7. Following a 24-hour incubation period and microscopic examination, the horizontal group exhibited greater osteoblast migratory activity than the control group and the fixed-angle centrifugation group. Horizontal centrifugation C-PRF had a significantly greater effect on osteoblast proliferation than fixed-angle centrifugation. The horizontal group's proliferation rate surpassed that of the fixed-angle group by up to 1.5 times on the third day of observation. Conclusion Compared to fixed-angle centrifugation, the horizontal method of preparing C-PRF has a greater effect on the release of growth factors and the behavior of osteoblasts. © 2025. The Author(s).