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Characteristics of autologous platelet rich plasma concentrates obtained by the buffy coat technique using different protocols

Nikoleta Ivanova, Galina Yaneva, Dobri Ivanov



In the last decade, platelet-rich concentrates, including the so-called platelet-rich plasma (PRP), have been gaining extreme popularity and are widely used in variety of clinical fields of medicine—dermatology, orthopedics, traumatology, plastic and reconstructive surgery, maxillofacial surgery, and others.


The aim of the article was to analyze the quantitative characteristics of the blood components in autologous platelet-rich plasma concentrates obtained by the buffy coat technique using different protocols.

Materials and Methods

Thirty participants (avg. age 42.8) were included in the present study. Venous blood (35 mL) was drawn from each participant—3 mL were separated for control group to determine average blood cell level. Four protocols for PRP obtaining (4 x 8 mL) were performed by using vacutainer tube with separating gel and monovette (S-Monovette) without a separating gel. The concentration factor (%) for each protocol was calculated relative to the average baseline blood values of the control group.


Protocol I showed increase in platelet concentrations by 91.55% and the leukocyte level was 107% higher compared to the control group. The second protocol (separating gel) demonstrated increase in platelet count by 337.06% and in leukocytes by 82.37%. The third protocol, based on double centrifugation, showed 352.08% increase in platelets and 40.54% decrease in white blood cell compared to controls. The fourth protocol demonstrated 389.84% increase in platelet level and 74.36% decrease in leukocyte number.


The use of separating gels into various techniques for obtaining PRP facilitates blood cell aspiration and makes the procedure faster and safer in ambulatory practice.


autologous platelet products; platelets; growth factors

Full Text


Bielecki T, Gazdzik TS, Szczepanski T. Re: "The effects of local platelet rich plasma delivery on diabetic fracture healing". What do we use: Platelet-rich plasma or platelet-rich gel? Bone. 2006;39(6):1388; author reply 1389. doi: 10.1016/j.bone.2006.06.015.

Cieslik-Bielecka A, Bielecki T, Gazdzik TS, Cieslik T, Szczepanski T. Improved treatment of mandibular odontogenic cysts with platelet-rich gel. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2008;105(4):423-9. doi: 10.1016/j.tripleo.2007.07.039.

Gandhi A, Doumas C, O'Connor JP, Parsons JR, Lin SS. The effects of local platelet rich plasma delivery on diabetic fracture healing. Bone. 2006;38(4):540-6. doi: 10.1016/j.bone.2005.10.019.

Kitoh H, Kitakoji T, Tsuchiya H, Mitsuyama H, Nakamura H, Katoh M, et al. Transplantation of marrow-derived mesenchymal stem cells and platelet-rich plasma during distraction osteogenesis-a preliminary result of three cases. Bone. 2004;35(4):892-8. doi: 10.1016/j.bone.2004.06.013.

Ivanov S. Cannulated screw fixation for dislocated fractures of the calcaneus. Scr Sci Med. 2019;51(2):23-9. doi: 10.14748/ssm.v51i1.5882.

Lowery GL, Kulkarni S, Pennisi AE. Use of autologous growth factors in lumbar spinal fusion. Bone. 1999;25(2 Suppl):47S-50S. doi: 10.1016/s8756-3282(99)00132-5.

Lynch SE, Buser D, Hernandez RA, Weber HP, Stich H, Fox CH, et al. Effects of the platelet-derived growth factor/insulin-like growth factor-i combination on bone regeneration around titanium dental implants. Results of a pilot study in beagle dogs. J Periodontol. 1991;62(11):710-6. doi: 10.1902/jop.1991.62.11.710.

Marx RE, Carlson ER, Eichstaedt RM, Schimmele SR, Strauss JE, Georgeff KR. Platelet-rich plasma: Growth factor enhancement for bone grafts. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 1998;85(6):638-46. doi: 10.1016/s1079-2104(98)90029-4.

Rutkowski JL, Fennell JW, Kern JC, Madison DE, Johnson DA. Inhibition of alveolar osteitis in mandibular tooth extraction sites using platelet-rich plasma. J Oral Impl. 2007;33(3):116-21. doi: 10.1563/1548-1336(2007)33[116:IOAOIM]2.0.CO;2.

Marx RE. Platelet-rich plasma (PRP): What is PRP and what is not PRP? Implant Dent. 2001;10(4):225-8. doi: 10.1097/00008505-200110000-00002.

Murphy S, Heaton WA, Rebulla P. Platelet production in the old world--and the new. Transfusion. 1996;36(8):751-4. doi: 10.1046/j.1537-2995.1996.36896374382.x

Dhurat Rachita and Sukesh MS. Principles and methods of preparation of platelet-rich plasma: A review and author's perspective. J Cutan Aesthet Surg. 2014;7(4):189–97. doi: 10.4103/0974-2077.150734.

Mazzucco L, Balbo V, Cattana E, Guaschino R, Borzini P. Not every PRP-gel is born equal. Evaluation of growth factor availability for tissues through four PRP-gel preparations: Fibrinet, RegenPRP-Kit, Plateltex and one manual procedure. Vox Sang. 2009;97(2):110-8. doi: 10.1111/j.1423-0410.2009.01188.x.

Margolis DJ, Kantor J, Santanna J, Strom BL, Berlin JA. Effectiveness of platelet releasate for the treatment of diabetic neuropathic foot ulcers. Diabetes Care. 2001;24(3):483-8. doi: 10.2337/diacare.24.3.483.

Hsu WK, Mishra A, Rodeo SR, Fu F, Terry MA, Randelli P. Platelet-rich plasma in orthopaedic applications: evidence-based recommendations for treatment. J Am Acad Orthop Surg. 2013;21(12):739-48. doi: 10.5435/JAAOS-21-12-739.

Kevy SV, Jacobson MS. Comparison of methods for point of care preparation of autologous platelet gel. J Extra Corpor Technol. 2004;36(1):28-35.

Slater M, Patava J, Kingham K, Mason RS. Involvement of platelets in stimulating osteogenic activity. J Orthop Res. 1995;13(5):655-63. doi: 10.1002/jor.1100130504.

Gurtner GC, Werner S, Barrandon Y, Longaker MT. Wound repair and regeneration. Nature. 2008;453(7193):314–21. doi: 10.1038/nature07039.

Berlanga-Acosta J, Gavilondo-Cowley J, Lopez-Saura P, Gonzalez-Lopez T, Castro-Santana MD, Lopez-Mola E, et al. Epidermal growth factor in clinical practice - a review of its biological actions, clinical indications and safety implications. Int Wound J. 2009;6(5):331-46. doi: 10.1111/j.1742-481X.2009.00622.x.

Marx RE. Platelet-rich plasma: evidence to support its use. J Oral Maxillofac Surg. 2004;62(4):489-96. doi: 10.1016/j.joms.2003.12.003.

Alsousou J, Thompson M, Hulley P, Noble A, Willett K. The biology of platelet-rich plasma and its application in trauma and orthopaedic surgery: a review of the literature. J Bone Joint Surg Br. 2009;91(8):987-96. doi: 10.1302/0301-620X.91B8.22546.

Cieslik-Bielecka A, Bielecki T, Gazdzik TS, Arendt J, Król W, Szczepanski T. Autologous platelets and leukocytes can improve healing of infected high-energy soft tissue injury. Transfus Apher Sci. 2009;41(1):9-12. doi: 10.1016/j.transci.2009.05.006.

McCarrel TM, Minas T, Fortier LA. Optimization of leukocyte concentration in platelet-rich plasma for the treatment of tendinopathy. J Bone Joint Surg Am. 2012;94(19):e143(1-8). doi: 10.2106/JBJS.L.00019.

Dragoo JL, Braun HJ, Durham JL, Ridley BA, Odegaard JI, Luong R, et al., Comparison of the acute inflammatory response of two commercial platelet-rich plasma systems in healthy rabbit tendons. Am J Sports Med. 2012:40(6):1274-81. doi: 10.1177/0363546512442334.

Dhillon RS, Schwarz EM, Maloney MD. Platelet-rich plasma therapy - future or trend? Arthritis Res Ther. 2012;14(4):219. doi: 10.1186/ar3914.

Lichtman MA, Beutler E, Kaushansky K, Kipps TJ, Seilgsohn U, Prchal J, editors. Williams hematology. 7th edition. New York McGraw-Hill; 2006.



About The Authors

Nikoleta Ivanova
Medical University of Varna

Department of Biology, Faculty of Pharmacy

Galina Yaneva
Medical University of Varna

Department of Biology, Faculty of Pharmacy

Dobri Ivanov
Medical University of Varna

Department of Biology, Faculty of Pharmacy

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