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Veterinary sciences2019; 6(3); doi: 10.3390/vetsci6030068

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.
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Publication Date: 2019-08-21 PubMed ID: 31438534PubMed Central: PMC6789863DOI: 10.3390/vetsci6030068Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

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, Fülber 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). https://doi.org/10.3390/vetsci6030068

Publication

Veterinary sciences
ISSN: 2306-7381
NlmUniqueID: 101680127
Country: Switzerland
Language: English
Volume: 6
Issue: 3

Researcher Affiliations

Seidel, Sarah R T
  • Departamento de Clínica Médica, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo 05508-270, Brazil. sarahseidel@usp.br.
Vendruscolo, Cynthia P
  • Departamento de Clínica Médica, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo 05508-270, Brazil.
Moreira, Juliana J
  • Departamento de Clínica Médica, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo 05508-270, Brazil.
Fülber, Joice
  • Departamento de Clínica Médica, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo 05508-270, Brazil.
Ottaiano, Tatiana F
  • Departamento de Bioquímica, Escola Paulista de Medicina, UNIFESP, São Paulo 04023-062, Brazil.
Oliva, Maria L V
  • Departamento de Bioquímica, Escola Paulista de Medicina, UNIFESP, São Paulo 04023-062, Brazil.
Michelacci, Yara M
  • Departamento de Bioquímica, Escola Paulista de Medicina, UNIFESP, São Paulo 04023-062, Brazil.
Baccarin, Raquel Y A
  • Departamento de Clínica Médica, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São 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
  1. Brossi PM, Moreira JJ, Machado TSL, Baccarin RYA. 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;11:98.
    doi: 10.1186/s12917-015-0403-zpmc: PMC4449579pubmed: 25896610google scholar: lookup
  2. Campbell KA, Saltzman BM, Mascarenhas R, Khair MM, Verma NN, Bach BR, 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;31:2213–2221.
    doi: 10.1016/j.arthro.2015.03.041pubmed: 26033459google scholar: lookup
  3. 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. Sport Traumatol. Arthrosc. 2015;23:2459–2474.
    doi: 10.1007/s00167-013-2743-1pmc: PMC4541701pubmed: 24275957google scholar: lookup
  4. Vendruscolo CP, Alves ALG, Brossi PM, Baccarin RYA. Uso do soro autólogo condicionado e do plasma rico em plaquetas na terapia ortopédica de equinos.. Semina 2014;35:2607–2624.
    doi: 10.5433/1679-0359.2014v35n5p2607google scholar: lookup
  5. Fortier LA, Barker JU, Strauss EJ, McCarrel TM, Cole BJ. The Role of Growth Factors in Cartilage Repair.. Clin. Orthop. Relat. Res. 2011;469:2706–2715.
    doi: 10.1007/s11999-011-1857-3pmc: PMC3171543pubmed: 21403984google scholar: lookup
  6. 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;54:103–110.
    doi: 10.1016/j.transci.2016.01.028pubmed: 26883885google scholar: lookup
  7. Argüelles D, Carmona JU, Pastor J, Iborra A, Viñals L, Martínez P, Bach P, Prades M. Evaluation of single and double centrifugation tube methods for concentrating equine platelets.. Res. Vet. Sci. 2006;81:237–245.
    doi: 10.1016/j.rvsc.2005.12.008pubmed: 16969921google scholar: lookup
  8. Sutter WW, Kaneps AJ, Bertone AL. Comparison of hematologic values and transforming growth factor-β 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;65:924–930.
    doi: 10.2460/ajvr.2004.65.924pubmed: 15281650google scholar: lookup
  9. Lubkowska A, Dolegowska B, Banfi G. Growth factor content in PRP and their applicability in medicine.. J. Biol. Regul. Homeost. Agents. 2012;26:3–22.
    pubmed: 23648195
  10. 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 β: Studies in anatomically intact cartilage in vitro and in vivo.. Ann. Rheum. Dis. 1993;52:185–191.
    doi: 10.1136/ard.52.3.185pmc: PMC1005015pubmed: 8484670google scholar: lookup
  11. 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.. Osteoarthr. Cartilage. 2009;17:1539–1545.
    doi: 10.1016/j.joca.2009.06.008pubmed: 19583961google scholar: lookup
  12. Freyria AM, Mallein-Gerin F. Chondrocytes or adult stem cells for cartilage repair: The indisputable role of growth factors.. Injury 2012;43:259–265.
    doi: 10.1016/j.injury.2011.05.035pubmed: 21696723google scholar: lookup
  13. Morales TI, Roberts AB. Transforming growth factor β regulates the metabolism of proteoglycans in bovine cartilage organ cultures.. J. Biol. Chem. 1988;263:12828–12831.
    pubmed: 3166454
  14. Imler SM, Doshi AN, Levenston ME. Combined effects of growth factors and static mechanical compression on meniscus explant biosynthesis.. Osteoarthr. Cartil. 2004;12:736–744.
    doi: 10.1016/j.joca.2004.05.007pubmed: 15325640google scholar: lookup
  15. Baugé C, Girard N, Lhuissier E, Bazille C, Boumediene K. Regulation and Role of TGFβ Signaling Pathway in Aging and Osteoarthritis Joints.. Aging Dis. 2014;5:394–405.
    doi: 10.14336/AD.2014.0500394pmc: PMC4249809pubmed: 25489490google scholar: lookup
  16. 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-β1.. Differentiation 1995;59:25–34.
    doi: 10.1046/j.1432-0436.1995.5910025.xpubmed: 7589892google scholar: lookup
  17. 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-β injections.. Osteoarthr. Cartil. 2000;8:25–33.
    doi: 10.1053/joca.1999.0267pubmed: 10607496google scholar: lookup
  18. Blaney Davidson EN, Van der Kraan PM, Van den Berg WB. TGF-β and osteoarthritis.. Osteoarthr. Cartil. 2007;15:597–604.
    doi: 10.1016/j.joca.2007.02.005pubmed: 17391995google scholar: lookup
  19. Fortier LA, Hackett CH, Cole BJ. The Effects of Platelet-Rich Plasma on Cartilage: Basic Science and Clinical Application.. Oper. Tech. Sports Med. 2011;19:154–159.
    doi: 10.1053/j.otsm.2011.03.004google scholar: lookup
  20. Floryan K, Berghoff W. Intraoperative use of autologous platelet-rich and platelet-poor plasma for orthopedic surgery patients.. AORN J. 2004;80:667–674.
    doi: 10.1016/S0001-2092(06)61320-3pubmed: 15526700google scholar: lookup
  21. 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;39:177–182.
    pmc: PMC4680728pubmed: 17972452
  22. 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. 2019;38:30–33.
    doi: 10.1016/j.foot.2018.11.003pubmed: 30572281google scholar: lookup
  23. 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. A. 2014;20:874–882.
    doi: 10.1089/ten.tea.2013.0058pmc: PMC3926147pubmed: 24098948google scholar: lookup
  24. Ottaiano TF, Andrade SS, De Oliveira C, Silva MCC, Buri MV, Juliano MA, Girão MJBC, Sampaio MU, Schmaier AH, Wlodawer A. Plasma kallikrein enhances platelet aggregation response by subthreshold doses of ADP.. Biochimie 2017;135:72–81.
    doi: 10.1016/j.biochi.2017.01.010pmc: PMC5346445pubmed: 28115185google scholar: lookup
  25. Born GVR, Cross MJ. The aggregation of blood platelets.. J. Physiol. 1963;168:178–195.
    doi: 10.1113/jphysiol.1963.sp007185pmc: PMC1359417pubmed: 14056485google scholar: lookup
  26. Tablin F, Walker NJ, Hogle SE, Pratt S, 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;69:1512–1519.
    doi: 10.2460/ajvr.69.11.1512pubmed: 18980435google scholar: lookup
  27. 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;198:217–223.
    doi: 10.1016/j.tvjl.2013.07.020pubmed: 23992870google scholar: lookup
  28. Fontenot RL, Sink CA, Were SR, Weinstein NM, Dahlgren LA. Simple tube centrifugation for processing platelet-rich plasma in the horse.. Can. Vet. J. 2012;53:1266–1272.
    pmc: PMC3500116pubmed: 23729823
  29. Vendruscolo CP, De Carvalho AM, Moraes LF, Maia L, Queiroz DL, Watanabe MJ, Yamada ALM, Alves ALG. Avaliação da eficácia de diferentes protocolos de preparo do Plasma Rico em Plaquetas para uso em Medicina Equina.. Pesqui. Vet. Bras. 2012;32:106–110.
    doi: 10.1590/S0100-736X2012000200002google scholar: lookup
  30. Hessel LN, Bosch G, Van Weeren PR, Ionita JC. Equine autologous platelet concentrates: A comparative study between different available systems.. Equine Vet. J. 2015;47:319–325.
    doi: 10.1111/evj.12288pubmed: 24773596google scholar: lookup
  31. Carmona JU, Argüelles 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;27:7–10.
    doi: 10.1016/j.jevs.2007.02.007google scholar: lookup
  32. 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;72:271–278.
    doi: 10.2460/ajvr.72.2.271pubmed: 21281204google scholar: lookup
  33. 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;27:1033–1042.
    doi: 10.1002/jor.20853pubmed: 19170097google scholar: lookup
  34. 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;39:2135–2140.
    doi: 10.1177/0363546511417792pubmed: 21846925google scholar: lookup
  35. Pereira RCDF, Zacarias GVF, Cantarelli C, Corrêa MMB, Silva GB, Barbosa ALT, Brass KE, De La Côrte FD. Avaliação De Sete Protocolos De Obtenção De Plasma Rico Em Plaquetas.. Ciência Rural. 2012;43:1122–1127.
    doi: 10.1590/S0103-84782013005000052google scholar: lookup
  36. 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;12:48–57.
  37. Textor JA, Tablin F. Activation of equine platelet-rich plasma: Comparison of methods and characterization of equine autologous thrombin.. Vet. Surg. 2012;41:784–794.
    doi: 10.1111/j.1532-950X.2012.01016.xpubmed: 22742830google scholar: lookup
  38. Lansdown DA, Fortier LA. Platelet Rich Plasma: Formulations, Preparations, Constituents, and Their Effects.. Oper. Tech. Sports Med. 2016;25:7–12.
    doi: 10.1053/j.otsm.2016.12.002google scholar: lookup
  39. Filardo G, Kon E, Pereira Ruiz MT, Vaccaro F, Guitaldi R, Di Martino A, Cennachi A, Fornasari PM, Marcacci M. Platelet-rich plasma intra-articular injections for cartilage degeneration and osteoarthritis: Single- versus double-spinning approach.. Knee Surg. Sport Traumatol. Arthrosc. 2012;20:2082–2091.
    doi: 10.1007/s00167-011-1837-xpubmed: 22203046google scholar: lookup
  40. 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;1:115–123.
    doi: 10.1089/biores.2012.0225pmc: PMC3559222pubmed: 23516671google scholar: lookup
  41. 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;62:547–554.
    doi: 10.2460/ajvr.2001.62.547pubmed: 11327462google scholar: lookup
  42. Segura D, Monreal L, Espada Y, Pastor J, Mayós I, Homedes J. Assessment of a platelet function analyser in horses: Reference range and influence of a platelet aggregation inhibitor.. Vet. J. 2005;170:108–112.
    doi: 10.1016/j.tvjl.2004.05.013pubmed: 15993794google scholar: lookup
  43. 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;39:729–734.
    doi: 10.1177/0363546511401576pmc: PMC3176726pubmed: 21398575google scholar: lookup

Citations

This article has been cited 9 times.
  1. Seidel SRT, Fülber J, Barbosa ÂP, Penatti NMA, Demasi M, Baccarin RYA. Medium-term storage of platelet-derived orthobiologics: a feasible alternative for equine practice. Front Vet Sci 2025;12:1720164.
    doi: 10.3389/fvets.2025.1720164pubmed: 41602624google scholar: lookup
  2. Carmona JU, López C, Argüelles D. Addressing Heterogeneity in Equine PRP Therapies: A Scoping Review of Methods, Evidence, and Commercial Validation. Animals (Basel) 2025 Dec 13;15(24).
    doi: 10.3390/ani15243586pubmed: 41463871google scholar: lookup
  3. Zhao YF, Pan Y, Liu HQ, Zhu FP, Wu Q. Differential Therapeutic Efficacy of Autologous Platelet-Rich Plasma at Varying Concentrations in the Treatment of Deep Second-Degree Burn and Its Underlying Molecular Mechanisms. J Cosmet Dermatol 2026 Jan;25(1):e70633.
    doi: 10.1111/jocd.70633pubmed: 41457485google scholar: lookup
  4. Yuan C, Ang SP, Hasoon JJ, Tolba R, Ruan QZ, Lam CM, Lo Bianco G, Christo PJ, Robinson CL. Dual-Action Regenerative Therapies: Regeneration and Antimicrobial Effects of Platelet- and Marrow-Derived Biologics. Biomedicines 2025 Nov 20;13(11).
    doi: 10.3390/biomedicines13112832pubmed: 41301922google scholar: lookup
  5. Mercader-Ruiz J, Beitia M, Delgado D, Sánchez P, Porras B, Gimeno I, González S, Benito-Lopez F, Basabe-Desmonts L, Sánchez M. Current Challenges in the Development of Platelet-Rich Plasma-Based Therapies. Biomed Res Int 2024;2024:6444120.
    doi: 10.1155/2024/6444120pubmed: 39157212google scholar: lookup
  6. 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.
    doi: 10.3389/fvets.2022.958724pubmed: 36090170google scholar: lookup
  7. Seidel SRT, de Souza AF, Fülber 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.
    pubmed: 34219782
  8. 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.
    doi: 10.1177/23259671211006100pubmed: 34159208google scholar: lookup
  9. 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).
    doi: 10.3390/pharmaceutics13010089pubmed: 33440840google scholar: lookup
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