Background: Full consensus optimizing the PRF preparation protocols could not be established yet. This study aimed to compare two commercially available centrifuges at various g and time protocols in terms of their influence on growth factor and cytokine values. Methods: Blood samples from 10 individuals were centrifuged either with PRF Duo® or Intra-Spin® device for PRF preparation. Values for PRF Duo® were adjusted to ~200g, ~400g, ~600g, ~800g and 10, 12, 14min while ~400g, 12min (control) was set for Intra-Spin®. PRF clots were embedded into well plates containing Dulbecco’s Modified Eagle’s Medium and PDGF-AB, TGF-β1 and IL-1β quantities at days 1, 3 and 7 were calculated by using ELISA kits. Results: Control and ~600g groups demonstrated prolonged PDGF-AB release compared to others whereas ~200g showed a tendency of burst release. A high and prolonged release pattern similar to control was detected in the 10min group. ~200g and ~600g groups showed high TGF-β1 release at day 1 and then, their release amounts reduced while an increase from day 1 to 3 was observed at only control group. Apart from the higher release at ~800g group, IL-1β release was similar for all study groups. Conclusions: In terms of growth factor and cytokine release, Intra-Spin® might be preferred instead of PRF Duo®. The recommended protocol of Intra-Spin® may give the best release outcomes for PRF Duo®. In order to confirm the results, further studies are needed.