Does Double Centrifugation Lead to Premature Platelet Aggregation and Decreased TGF-β1 Concentrations in Equine Platelet-Rich Plasma?
Abstract: Blood-derived autologous products are frequently used in both human and equine medicine to treat musculoskeletal disorders. These products, especially the platelet-rich plasma (PRP), may contain high concentrations of growth factors (GFs), and thus improve healing in several tissues. Nevertheless, the procedures for preparation of PRP are currently non-standardized. Several protocols, which are based on distinct centrifugation patterns (rotation speed and time), result in PRPs with different characteristics, concerning platelet and GFs concentrations, as well as platelet activation. The aim of the present study was to compare two different protocols for PRP preparation: protocol (A) that is based on a single-centrifugation step; protocol (B), which included two sequential centrifugation steps (double-centrifugation). The results here reported show that the double-centrifugation protocol resulted in higher platelet concentration, while leukocytes were not concentrated by this procedure. Although platelet activation and aggregation were increased in this protocol in comparison to the single-centrifugation one, the TGF-β1 concentration was also higher. Pearson's correlation coefficients gave a significant, positive correlation between the platelet counts and TGF-β1 concentration. In conclusion, although the double-centrifugation protocol caused premature platelet aggregation, it seems to be an effective method for preparation of PRP with high platelet and TGF-β1 concentrations.
Publication Date: 2019-08-21 PubMed ID: 31438534PubMed Central: PMC6789863DOI: 10.3390/vetsci6030068Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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This research examines the effectiveness of two different preparation methods for platelet-rich plasma (PRP), a treatment used in human and equine medicine for musculoskeletal disorders. It indicates that the double-centrifugation protocol, although leading to premature platelet aggregation, resulted in a higher concentration of platelets and TGF-β1 compared to the single-centrifugation method.
Introduction and Aim
- The researchers aimed to compare two different protocols for preparing PRP: a single-centrifugation step (protocol A) and a double-centrifugation step (protocol B).
- PRP, frequently used in human and equine medicine to treat musculoskeletal disorders, is important for its high concentration of growth factors which improves tissue healing.
- The researchers note a lack of standardization in PRP preparation procedures, with different centrifugation patterns leading to varying results in platelet activation and growth factor concentration.
Results and Analysis
- The study findings indicated that protocol B, the double-centrifugation method, produced a higher platelet concentration than protocol A.
- This protocol did not result in a concentration of leukocytes, white cells involved in protecting the body against infectious disease and foreign invaders.
- Despite causing increased platelet activation and aggregation, this method also resulted in a higher TGF-β1 concentration, a type of growth factor.
- The correlation analysis (Pearson’s correlation) showed a significant positive correlation between platelet counts and TGF-β1 concentration, suggesting that the more platelets, the higher the TGF-β1 concentration.
Conclusion
- While acknowledging that the double-centrifugation method does influence premature platelet aggregation, the researchers concluded it as an effective method for creating PRP with high concentrations of platelets and TGF-β1.
- This could have implications for the efficiency of PRP preparation and its effectiveness in treating patients, given the importance of platelets and growth factors in tissue healing.
Cite This Article
APA
Seidel SRT, Vendruscolo CP, Moreira JJ, Fu00fclber J, Ottaiano TF, Oliva MLV, Michelacci YM, Baccarin RYA.
(2019).
Does Double Centrifugation Lead to Premature Platelet Aggregation and Decreased TGF-β1 Concentrations in Equine Platelet-Rich Plasma?
Vet Sci, 6(3), 68.
https://doi.org/10.3390/vetsci6030068 Publication
Researcher Affiliations
- Departamento de Clu00ednica Mu00e9dica, Faculdade de Medicina Veterinu00e1ria e Zootecnia, Universidade de Su00e3o Paulo, Su00e3o Paulo 05508-270, Brazil. sarahseidel@usp.br.
- Departamento de Clu00ednica Mu00e9dica, Faculdade de Medicina Veterinu00e1ria e Zootecnia, Universidade de Su00e3o Paulo, Su00e3o Paulo 05508-270, Brazil.
- Departamento de Clu00ednica Mu00e9dica, Faculdade de Medicina Veterinu00e1ria e Zootecnia, Universidade de Su00e3o Paulo, Su00e3o Paulo 05508-270, Brazil.
- Departamento de Clu00ednica Mu00e9dica, Faculdade de Medicina Veterinu00e1ria e Zootecnia, Universidade de Su00e3o Paulo, Su00e3o Paulo 05508-270, Brazil.
- Departamento de Bioquu00edmica, Escola Paulista de Medicina, UNIFESP, Su00e3o Paulo 04023-062, Brazil.
- Departamento de Bioquu00edmica, Escola Paulista de Medicina, UNIFESP, Su00e3o Paulo 04023-062, Brazil.
- Departamento de Bioquu00edmica, Escola Paulista de Medicina, UNIFESP, Su00e3o Paulo 04023-062, Brazil.
- Departamento de Clu00ednica Mu00e9dica, Faculdade de Medicina Veterinu00e1ria e Zootecnia, Universidade de Su00e3o Paulo, Su00e3o Paulo 05508-270, Brazil. baccarin@usp.br.
Conflict of Interest Statement
The authors declare that have no conflict of interest.
References
This article includes 43 references
- Brossi PM, Moreira JJ, Machado TS, Baccarin RY. Platelet-rich plasma in orthopedic therapy: a comparative systematic review of clinical and experimental data in equine and human musculoskeletal lesions.. BMC Vet Res 2015 Apr 22;11:98.
- Campbell KA, Saltzman BM, Mascarenhas R, Khair MM, Verma NN, Bach BR Jr, Cole BJ. Does Intra-articular Platelet-Rich Plasma Injection Provide Clinically Superior Outcomes Compared With Other Therapies in the Treatment of Knee Osteoarthritis? A Systematic Review of Overlapping Meta-analyses.. Arthroscopy 2015 Nov;31(11):2213-21.
- Filardo G, Kon E, Roffi A, Di Matteo B, Merli ML, Marcacci M. Platelet-rich plasma: why intra-articular? A systematic review of preclinical studies and clinical evidence on PRP for joint degeneration.. Knee Surg Sports Traumatol Arthrosc 2015 Sep;23(9):2459-74.
- Vendruscolo C.P., Alves A.L.G., Brossi P.M., Baccarin R.Y.A. Uso do soro autu00f3logo condicionado e do plasma rico em plaquetas na terapia ortopu00e9dica de equinos. Semina. 2014;35:2607u20132624. doi: 10.5433/1679-0359.2014v35n5p2607.
- Fortier LA, Barker JU, Strauss EJ, McCarrel TM, Cole BJ. The role of growth factors in cartilage repair.. Clin Orthop Relat Res 2011 Oct;469(10):2706-15.
- Arora S, Doda V, Kotwal U, Dogra M. Quantification of platelets and platelet derived growth factors from platelet-rich-plasma (PRP) prepared at different centrifugal force (g) and time.. Transfus Apher Sci 2016 Feb;54(1):103-10.
- Argu00fcelles D, Carmona JU, Pastor J, Iborra A, Viu00f1als L, Martu00ednez P, Bach E, Prades M. Evaluation of single and double centrifugation tube methods for concentrating equine platelets.. Res Vet Sci 2006 Oct;81(2):237-45.
- Sutter WW, Kaneps AJ, Bertone AL. Comparison of hematologic values and transforming growth factor-beta and insulin-like growth factor concentrations in platelet concentrates obtained by use of buffy coat and apheresis methods from equine blood.. Am J Vet Res 2004 Jul;65(7):924-30.
- Lubkowska A, Dolegowska B, Banfi G. Growth factor content in PRP and their applicability in medicine.. J Biol Regul Homeost Agents 2012 Apr-Jun;26(2 Suppl 1):3S-22S.
- van Beuningen HM, van der Kraan PM, Arntz OJ, van den Berg WB. Protection from interleukin 1 induced destruction of articular cartilage by transforming growth factor beta: studies in anatomically intact cartilage in vitro and in vivo.. Ann Rheum Dis 1993 Mar;52(3):185-91.
- van der Kraan PM, Blaney Davidson EN, Blom A, van den Berg WB. TGF-beta signaling in chondrocyte terminal differentiation and osteoarthritis: modulation and integration of signaling pathways through receptor-Smads.. Osteoarthritis Cartilage 2009 Dec;17(12):1539-45.
- Freyria AM, Mallein-Gerin F. Chondrocytes or adult stem cells for cartilage repair: the indisputable role of growth factors.. Injury 2012 Mar;43(3):259-65.
- Morales TI, Roberts AB. Transforming growth factor beta regulates the metabolism of proteoglycans in bovine cartilage organ cultures.. J Biol Chem 1988 Sep 15;263(26):12828-31.
- Imler SM, Doshi AN, Levenston ME. Combined effects of growth factors and static mechanical compression on meniscus explant biosynthesis.. Osteoarthritis Cartilage 2004 Sep;12(9):736-44.
- Baugu00e9 C, Girard N, Lhuissier E, Bazille C, Boumediene K. Regulation and Role of TGFu03b2 Signaling Pathway in Aging and Osteoarthritis Joints.. Aging Dis 2014 Dec;5(6):394-405.
- Denker AE, Nicoll SB, Tuan RS. Formation of cartilage-like spheroids by micromass cultures of murine C3H10T1/2 cells upon treatment with transforming growth factor-beta 1.. Differentiation 1995 Jul;59(1):25-34.
- van Beuningen HM, Glansbeek HL, van der Kraan PM, van den Berg WB. Osteoarthritis-like changes in the murine knee joint resulting from intra-articular transforming growth factor-beta injections.. Osteoarthritis Cartilage 2000 Jan;8(1):25-33.
- Blaney Davidson EN, van der Kraan PM, van den Berg WB. TGF-beta and osteoarthritis.. Osteoarthritis Cartilage 2007 Jun;15(6):597-604.
- Fortier L.A., Hackett C.H., Cole B.J. The Effects of Platelet-Rich Plasma on Cartilage: Basic Science and Clinical Application. Oper. Tech. Sports Med. 2011;19:154u2013159. doi: 10.1053/j.otsm.2011.03.004.
- Floryan KM, Berghoff WJ. Intraoperative use of autologous platelet-rich and platelet-poor plasma for orthopedic surgery patients.. AORN J 2004 Oct;80(4):668-74, quiz 675-8.
- Zavadil DP, Satterlee CC, Costigan JM, Holt DW, Shostrom VK. Autologous platelet gel and platelet-poor plasma reduce pain with total shoulder arthroplasty.. J Extra Corpor Technol 2007 Sep;39(3):177-82.
- Malahias MA, Mavrogenis AF, Nikolaou VS, Megaloikonomos PD, Kazas ST, Chronopoulos E, Babis GC. Similar effect of ultrasound-guided platelet-rich plasma versus platelet-poor plasma injections for chronic plantar fasciitis.. Foot (Edinb) 2019 Mar;38:30-33.
- Hatakeyama I, Marukawa E, Takahashi Y, Omura K. Effects of platelet-poor plasma, platelet-rich plasma, and platelet-rich fibrin on healing of extraction sockets with buccal dehiscence in dogs.. Tissue Eng Part A 2014 Feb;20(3-4):874-82.
- Ottaiano TF, Andrade SS, de Oliveira C, Silva MCC, Buri MV, Juliano MA, Giru00e3o MJBC, Sampaio MU, Schmaier AH, Wlodawer A, Maffei FHA, Oliva MLV. Plasma kallikrein enhances platelet aggregation response by subthreshold doses of ADP.. Biochimie 2017 Apr;135:72-81.
- BORN GV, CROSS MJ. THE AGGREGATION OF BLOOD PLATELETS.. J Physiol 1963 Aug;168(1):178-95.
- Tablin F, Walker NJ, Hogle SE, Pratt SM, Norris JW. Assessment of platelet growth factors in supernatants from rehydrated freeze-dried equine platelets and their effects on fibroblasts in vitro.. Am J Vet Res 2008 Nov;69(11):1512-9.
- Textor JA, Willits NH, Tablin F. Synovial fluid growth factor and cytokine concentrations after intra-articular injection of a platelet-rich product in horses.. Vet J 2013 Oct;198(1):217-23.
- Fontenot RL, Sink CA, Werre SR, Weinstein NM, Dahlgren LA. Simple tube centrifugation for processing platelet-rich plasma in the horse.. Can Vet J 2012 Dec;53(12):1266-72.
- Vendruscolo C.P., De Carvalho A.M., Moraes L.F., Maia L., Queiroz D.L., Watanabe M.J., Yamada A.L.M., Alves A.L.G. Avaliau00e7u00e3o da eficu00e1cia de diferentes protocolos de preparo do Plasma Rico em Plaquetas para uso em Medicina Equina. Pesqui. Vet. Bras. 2012;32:106u2013110. doi: 10.1590/S0100-736X2012000200002.
- Hessel LN, Bosch G, van Weeren PR, Ionita JC. Equine autologous platelet concentrates: A comparative study between different available systems.. Equine Vet J 2015 May;47(3):319-25.
- Carmona J.U., Argu00fcelles D., Climent F., Prades M. Autologous Platelet Concentrates as a Treatment of Horses with Osteoarthritis: A Preliminary Pilot Clinical Study. J. Equine Vet. Sci. 2007;27:7u201310. doi: 10.1016/j.jevs.2007.02.007.
- Textor JA, Norris JW, Tablin F. Effects of preparation method, shear force, and exposure to collagen on release of growth factors from equine platelet-rich plasma.. Am J Vet Res 2011 Feb;72(2):271-8.
- McCarrel T, Fortier L. Temporal growth factor release from platelet-rich plasma, trehalose lyophilized platelets, and bone marrow aspirate and their effect on tendon and ligament gene expression.. J Orthop Res 2009 Aug;27(8):1033-42.
- Sundman EA, Cole BJ, Fortier LA. Growth factor and catabolic cytokine concentrations are influenced by the cellular composition of platelet-rich plasma.. Am J Sports Med 2011 Oct;39(10):2135-40.
- Pereira R.C.D.F., Zacarias G.V.F., Cantarelli C., Corru00eaa M.M.B., Silva G.B., Barbosa A.L.T., Brass K.E., De La Cu00f4rte F.D. Avaliau00e7u00e3o De Sete Protocolos De Obtenu00e7u00e3o De Plasma Rico Em Plaquetas. Ciu00eancia Rural. 2012;43:1122u20131127. doi: 10.1590/S0103-84782013005000052.
- McLellan J., Plevin S. Temporal release of growth factors from platelet-rich Fibrin (PRF) and Platelet-rich Rlasma (PRP) in the horse: A comparative in vitro analysis. Int. J. Appl. Res. Vet. Med. 2014;12:48u201357.
- Textor JA, Tablin F. Activation of equine platelet-rich plasma: comparison of methods and characterization of equine autologous thrombin.. Vet Surg 2012 Oct;41(7):784-94.
- Lansdown D.A., Fortier L.A. Platelet Rich Plasma: Formulations, Preparations, Constituents, and Their Effects. Oper. Tech. Sports Med. 2016;25:7u201312. doi: 10.1053/j.otsm.2016.12.002.
- Filardo G, Kon E, Pereira Ruiz MT, Vaccaro F, Guitaldi R, Di Martino A, Cenacchi A, Fornasari PM, Marcacci M. Platelet-rich plasma intra-articular injections for cartilage degeneration and osteoarthritis: single- versus double-spinning approach.. Knee Surg Sports Traumatol Arthrosc 2012 Oct;20(10):2082-91.
- Bausset O, Giraudo L, Veran J, Magalon J, Coudreuse JM, Magalon G, Dubois C, Serratrice N, Dignat-George F, Sabatier F. Formulation and storage of platelet-rich plasma homemade product.. Biores Open Access 2012 Jun;1(3):115-23.
- Kingston JK, Bayly WM, Sellon DC, Meyers KM, Wardrop KJ. Effects of sodium citrate, low molecular weight heparin, and prostaglandin E1 on aggregation, fibrinogen binding, and enumeration of equine platelets.. Am J Vet Res 2001 Apr;62(4):547-54.
- Segura D, Monreal L, Espada Y, Pastor J, Mayu00f3s I, Homedes J. Assessment of a platelet function analyser in horses: reference range and influence of a platelet aggregation inhibitor.. Vet J 2005 Jul;170(1):108-12.
- Harrison S, Vavken P, Kevy S, Jacobson M, Zurakowski D, Murray MM. Platelet activation by collagen provides sustained release of anabolic cytokines.. Am J Sports Med 2011 Apr;39(4):729-34.
Citations
This article has been cited 4 times.- Even KM, Gaesser AM, Ciamillo SA, Linardi RL, Ortved KF. Comparing the immunomodulatory properties of equine BM-MSCs culture expanded in autologous platelet lysate, pooled platelet lysate, equine serum and fetal bovine serum supplemented culture media.. Front Vet Sci 2022;9:958724.
- Seidel SRT, de Souza AF, Fu00fclber J, Bogossian PM, Rodrigues NNP, Baccarin RYA. Evaluation of platelet-rich plasma applied in the coronary band of healthy equine hooves.. Can Vet J 2021 Jul;62(7):729-735.
- Liu B, Jeong HJ, Yeo JH, Oh JH. Efficacy of Intraoperative Platelet-Rich Plasma Augmentation and Postoperative Platelet-Rich Plasma Booster Injection for Rotator Cuff Healing: A Randomized Controlled Clinical Trial.. Orthop J Sports Med 2021 Jun;9(6):23259671211006100.
- Bianchi E, Ruggeri M, Rossi S, Vigani B, Miele D, Bonferoni MC, Sandri G, Ferrari F. Innovative Strategies in Tendon Tissue Engineering.. Pharmaceutics 2021 Jan 11;13(1).