A Cycle of Freezing and Thawing as a Modified Method for Activating Platelets in Platelet-rich Plasma to Use in Regenerative Medicine
İbrahim Eker, Soner Yilmaz, Rıza A Çetinkaya, Aytekin Ünlü, Aysel Pekel, Rahşan I Sağkan, Zerrin Ertaş, Orhan Gürsel, Hacı U Muşabak, Ertuğrul Yazici, Sebahattin Yilmaz, Cengizhan Açikel, İsmail Y Avci, Ferit Avcu, Emin Kürekçi, Patrizio Petrone
Citation Information :
Eker İ, Yilmaz S, Çetinkaya RA, Ünlü A, Pekel A, Sağkan RI, Ertaş Z, Gürsel O, Muşabak HU, Yazici E, Yilmaz S, Açikel C, Avci İY, Avcu F, Kürekçi E, Petrone P. A Cycle of Freezing and Thawing as a Modified Method for Activating Platelets in Platelet-rich Plasma to Use in Regenerative Medicine. Panam J Trauma Crit Care Emerg Surg 2020; 9 (2):101-104.
Background: Platelet-rich plasma (PRP) has been widely used to improve wound healing and tissue repair. The objective of this study was to assess a cycle of freezing/thawing and its effect in the amount of growth factors in PRP. Materials and methods: After the preparation of PRP using standard methods, the samples were equally divided into two groups: The first group was activated by adding 10% calcium gluconate, while the second group was cryopreserved at −80°C for 24 hours. Enzyme-linked immunosorbent assay was used for aliquots of PRP to measure concentrations of insulin-like growth factor-1, platelet-derived growth factor, and basic fibroblast growth factor. Results: 15 mL of whole blood was obtained from 20 volunteers and collected into citrated tubes. The mean platelet count of the donors and the autologous PRP were 238.5 ± 44.7 × 103/μL and 544.7 ± 161.5 × 103/μL, respectively. There were no significant differences between the growth factor levels of freeze-thawed and calcium-activated PRP. A cycle of freezing/thawing was the only independent factor associated with the growth factor yield in the multivariate model. Conclusion: With its features of being simple, inexpensive, and easy for standardization, a cycle of freezing/thawing may be the method of choice for PRP activation procedure, without inducing fibrin matrix. This study was performed in a laboratory setting; therefore, future clinical trials are recommended.
Nurden AT, Nurden P, Sanchez M, et al. Platelets and wound healing. Front Biosci 2008;13(13):3532–3548. DOI: 10.2741/2947.
Blair P, Flaumenhaft R. Platelet alpha-granules: basic biology and clinical correlates. Blood Rev 2009;23(4):177–189. DOI: 10.1016/j.blre.2009.04.001.
Montmayeur JP, Valius M, Vandenheede J, et al. The platelet-derived growth factor beta receptor triggers multiple cytoplasmic signaling cascades that arrive at the nucleus as distinguishable inputs. J Biol Chem 1997;272(51):32670–32678. DOI: 10.1074/jbc.272.51.32670.
De Pascale MR, Sommese L, Casamassimi A, et al. Platelet derivatives in regenerative medicine: an update. Transfus Med Rev 2015;29(1): 52–61. DOI: 10.1016/j.tmrv.2014.11.001.
Cetinkaya RA, Yilmaz S, Ünlü A, et al. The efficacy of platelet-rich plasma gel in MRSA-related surgical wound infection treatment: an experimental study in an animal model. Eur J Trauma Emerg Surg 2018;44(6):859–867. DOI: 10.1007/s00068-017-0852-0.
Çetinkaya RA, Yenilmez E, Petrone P, et al. Platelet-rich plasma as an additional therapeutic option for infected wounds with multi-drug resistant bacteria: in vitro antibacterial activity study. Eur J Trauma Emerg Surg 2019;45(3):555–565. DOI: 10.1007/s00068-018- 0957-0.
Russell RP, Apostolakos J, Hirose T, et al. Variability of platelet-rich plasma preparations. Sports Med Arthrosc Rev 2013;21(4):186–190. DOI: 10.1097/JSA.0000000000000007.
Marques LF, Stessuk T, Camargo IC, et al. Platelet-rich plasma (PRP): methodological aspects and clinical applications. Platelets 2015;26(2):101–113. DOI: 10.3109/09537104.2014.881991.
Moshiri A, Oryan A. Role of platelet rich plasma in soft and hard connective tissue healing: an evidence based review from basic to clinical application. Hard Tissue 2013;2(1):6. DOI: 10.13172/2050-2303-2-1-326.
Zhu Y, Yuan M, Meng HY, et al. Basic science and clinical application of platelet-rich plasma for cartilage defects and osteoarthritis: a review. Osteoarthritis Cartilage 2013;21(11):1627–1637. DOI: 10.1016/j.joca.2013.07.017.
Holme S. Storage and quality assessment of platelets. Vox Sang 1998;74(Suppl 2):207–216. DOI: 10.1111/j.1423-0410.1998. tb05422.x.
Barrientos S, Stojadinovic O, Golinko MS, et al. Growth factors and cytokines in wound healing. Wound Repair Regen 2008;16(5):585–601. DOI: 10.1111/j.1524-475X.2008.00410.x.
Sekido Y, Morishima Y, Ohya K. Activity of platelet-derived growth factor (PDGF) in platelet concentrates and cryopreserved platelets determined by PDGF bioassay. Vox Sang 1987;52(1-2):27–30. DOI: 10.1111/j.1423-0410.1987.tb02984.x.
Roffi A, Filardo G, Assirelli E, et al. Does platelet-rich plasma freeze-thawing influence growth factor release and their effects on chondrocytes and synoviocytes? Biomed Res Int 2014;2014:692913. DOI: 10.1155/2014/692913.
Ronci C, Ferraro AS, Lanti A, et al. Platelet-rich plasma as treatment for persistent ocular epithelial defects. Transfus Apher Sci 2015;52(3):300–304. DOI: 10.1016/j.transci.2014.12.027.
Weed B, Davis MDP, Felty CL, et al. Autologous platelet lysate product versus placebo in patients with chronic leg ulcerations: a pilot study using a randomized, double-blind, placebo-controlled trial. Wounds 2004;16(9):273–282.
Sonker A, Dubey A. Determining the effect of preparation and storage: an effort to streamline platelet components as a source of growth factors for clinical application. Transfus Med Hemother 2015;42(3):174–180. DOI: 10.1159/000371504.