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.
Methods and Materials: 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.


  1. Chow TW, McIntire LV, Peterson DM. Importance of plasma fibronectin in determining PFP and PRP clot mechanical properties. Thromb Res. 1983;29:243-4.
  2. Delaini F, Poggi A, Donati MB. Enhanced affinity for arachidonic acid in platelet-rich plasma from rats with adriamycin-induced nephrotic syndrome. Thromb Haemost. 1982;48:260-2.
  3. Anfossi G, Trovati M, Mularoni E, Massucco P, Calcamuggi G, G Emanuelli. Influence of propranolol on platelet aggregation and thromboxane B2 production from platelet-rich plasma and whole blood. Prostaglandins Leukot Essent Fatty Acids. 1989;36:1-7.
  4. Singh B, Lynne JG. Autologous platelet-rich plasma for the treatment of pattern hair loss. Am J Clin Dermatol. 2016;17:359-67.
  5. Miron RJ, Fujioka-Kobayashi M, Bishara M, Zhang Y, Hernandez M, Choukroun J. Platelet-rich fibrin and soft tissue wound healing: A systematic review. Tissue Eng Part B Rev. 2017;23:83-99.
  6. Choukroun J, Adda F, Schoeffer C, Vervelle A. PRF: An opportunity in perio-implantology. Implantodontie. 2000;42:55-62.
  7. Ajwani H, Shetty S, Gopalakrishnan D, Kathariya R, Kulloli A, Dolas RS, et al. Comparative evaluation of platelet-rich fibrin biomaterial and open flap debridement in the treatment of two and three wall intrabony defects. J Int Oral Health. 2015;7:32-7.
  8. Choukron J, Adda F, Schoeffler C, Vervelle A. The opportunities en paro implantologie: Le PRF. Implantodontie. 2001;42:55-62.
  9. Femminella B, Iaconi MC, Di Tullio M, Romano L, Sinjari B, D'Arcangelo C, et al. Clinical comparison of platelet-rich fibrin and a gelatin sponge in the management of palatal wounds after epithelialized free gingival graft harvest: A randomized clinical trial. J Periodontol. 2016;87:103-13.
  10. Jain V, Triveni MG, Kumar AB, Mehta DS. Role of platelet-rich-fibrin in enhancing palatal wound healing after free graft. Contemp Clin Dent. 2012;3:S240-3.
  11. Sammartino G, Ehrenfest DM, Carile F, Tia M, Bucci P. Prevention of hemorrhagic complications after dental extractions into open heart surgery patients under anticoagulant therapy: The use of leukocyte-and platelet-rich fibrin. J Oral Implantol. 2011;37:681-90.
  12. Miron RJ, Fujioka-Kobayashi M, Hernandez M, Kandalam U, Zhang Y, Ghanaati S, et al. Injectable platelet rich fibrin (i-PRF): Opportunities in regenerative dentistry? Clin Oral Investig. 2017;21:2619-27.
  13. Kobayashi E, Flückiger L, Fujioka-Kobayashi M, Sawada K, Sculean A, Schaller B, et al. Comparative release of growth factors from PRP, PRF, and advanced-PRF. Clin Oral Investig. 2016;20:2353-60.
  14. Mourao CF, Valiense H, Melo ER, Mourão NB, Maia MD. Obtention of injectable platelets rich-fibrin (i-PRF) and its polymerization with bone graft: Technical Note. Rev Col Bras Cir. 2015;42:421-3.
  15. Al-Maawi S, Vorakulpipat C, Orlowska A, Zrnc TA, Sader RA, Kirkpatrick CJ, et al. In vivo implantation of a bovine-derived collagen membrane leads to changes in the physiological cellular pattern of wound healing by the induction of multinucleated giant cells: An adverse reaction? Front Bioeng Biotechnol. 2018;6:104.
  16. Castro AB, Cortellini S, Temmerman A, Li X, Pinto N, Teughels W, et al. Characterization of the leukocyte-and platelet-rich fibrin block: Release of growth factors, cellular content, and structure. Int J Oral Maxillofac Implants. 2019;34:855-64.
  17. Cortellini S, Castro AB, Temmerman A, van Dessel J, Pinto N, Jacobs R, et al. Leucocyte-and platelet-rich fibrin block for bone augmentation procedure: A proof-of-concept study. J Clin Periodontol. 2018;45:624-34.
  18. Miron RJ, Chai J, Zheng S, Feng M, Sculean A, Zhang Y. A novel method for evaluating and quantifying cell types in platelet rich fibrin and an introduction to horizontal centrifugation. J Biomed Mater Res A. 2019;107:2257-71.
  19. Lourenco ES, Mourão CF, Leite PE, Granjeiro JM, CalasansMaia MD, Alves GG. The in vitro release of cytokines and growth factors from fibrin membranes produced through horizontal centrifugation. J Biomed Mater Res A. 2018;106:1373-80.
  20. Nunes CR, Roedersheimer MT, Simske SJ, Luttges MW. Effect of microgravity, temperature, and concentration on fibrin and collagen assembly. Microgravity Sci Technol. 1995;8:125-30.
  21. Miron RJ, Chai J, Fujioka-Kobayashi M, Sculean A, Zhang Y. Evaluation of 24 protocols for the production of platelet-rich fibrin. BMC Oral Health. 2020;20:310.
  22. Fitzpatrick J, Bulsara MK, McCrory PR, Richardson MD, Zheng MH. Analysis of platelet-rich plasma extraction: Variations in platelet and blood components between 4 common commercial kits. Orthop J Sports Med. 2017;5:232596711667527.
  23. Moojen DJ, Everts PA, Schure RM, Overdevest EP, van Zundert A, Knape JT, et al. Antimicrobial activity of platelet-leukocyte gel against Staphylococcus aureus. J Orthop Res. 2008;26:404-10.
  24. Sundman EA, Cole BJ, Fortier A. Growth factor and catabolic cytokine concentrations are influenced by the cellular composition of platelet-rich plasma. Am J Sports Med. 2011;39:2135-40.
  25. Filardo G, Kon E, Ruiz MT, Vaccaro F, Guitaldi R, Di Martino A, et al. Platelet-rich plasma intra-articular injections for cartilage degeneration and osteoarthritis: Single-versus double-spinning approach. Knee Surg Sports Traumatol Arthrosc. 2012;20:2082-91.
  26. Kenmochi M. Clinical outcomes following injections of leukocyte-rich platelet-rich plasma in osteoarthritis patients. J Orthop. 2019;18:143-9.
  27. Mariani E, Canella V, Cattini L, Kon E, Marcacci M, Di Matteo B, et al. Leukocyte-rich platelet-rich plasma injections do not up-modulate intra-articular pro-inflammatory cytokines in the osteoarthritic knee. PLoS One. 2016;11:e0156137.

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