Objectives: Platelet-rich fibrin (PRF) is an autogenous biomaterial, considered as the second-generation platelet concentrates rich in blood cells and growth factors entrapped in the fibrin matrix, which makes it as an ideal material with wound healing abilities. Studies have reported high harvest of cells using anticoagulants but the present study employs two different protocols to efficiently separate the platelet-rich layer at low centrifugal forces without the use of anticoagulants. Material and Methods: Blood samples were collected with the consent of 20 volunteer donors. Ten blood samples were tested in each of the protocols studied, that is, protocol 1–200 g for 8 min (higher rpm and time) and protocol 2–60 g for 5 min (lower rpm and time). From the 12 ml of blood samples collected, 2 ml of blood was utilized for whole blood study; the remaining 10 ml was transferred into falcon tubes subjected to different rpm. Results: The present study employs a novel method to investigate segregation of cell types following low-grade centrifugation. One milliliter sequential pipetting technique was used to analyze number of leukocytes and platelets following centrifugation at two different g-forces. The protocols followed in the present study had 2–3-fold increase leukocytes concentration and 10–12-fold increases in platelet concentrations in the layers than the whole blood sample without the use of anticoagulants. Conclusion: The study concludes that protocol 1 was more efficient in harvesting platelets with less concentration of leukocytes, which is considered more suitable for various medical applications.
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