Influencing factors for preparation of platelet-rich plasma in horses

Ana Carolina Barros da Rosa Pedroso, Andréia da Costa Peixoto, Evelyn de Oliveira, Helena Tavares Dutra, Roberta Carvalho Basile, Luciana Ramos Gaston Brandstetter, Veridiana Maria Brianezi Dignani de Moura


There is no consensus in the literature as to the best platelet-rich plasma (PRP) acquisition method or the ideal platelet concentration to stimulate tissue repair. Besides that, most studies do not provide a clear and replicable description of the methods used, which makes standardization and result comparison difficult. Thus, this study aimed to accurately describe the method and evaluate factors influencing PRP preparation in equines. In this regard, two protocols were used, P1 and P2, based on two centrifugation methods, which differed in blood volume, speed and time, and platelet pipetting and resuspension. In conclusion, factors such as centrifugation strength and time are essential to obtain PRP with adequate platelet count. However, other factors such as tube type and plasma pipetting and resuspension methods directly interfere with the replicability of the technique, and hence influencing PRP applicability.


Platelet concentration; Double centrifugation.;Standardization.

Full Text:



Anitua, E., Andí, I., Sanchez, M., Azofra, J., del Mar Zalduendo, M., de la Fuente, M., & Nurden, A. T. (2005). Autologous preparations rich in growth factors promote proliferation and induce VEGF and HGF production by human tendon cells in culture. Journal of Orthopaedic Research, 23(2), 281-286. doi: 10.1016/j.orthres.2004.08.015

Anitua, E., Aguirre, J. J., Algorta, J., Ayerdi, E., Cabezas, A. I., Orive, G., & Andia, I. (2008). Effectiveness of autologous preparation rich in growth factors for the treatment of chronic cutaneous ulcers. Journal of Biomedical Materials Research Part B: Applied Biomaterials: An Official Journal of The Society for Biomaterials, The Japanese Society for Biomaterials, and The Australian Society for Biomaterials and the Korean Society for Biomaterials, 84(2), 415-421. doi: 10.1002/jbm.b.30886

Broggini, N., Hofstetter, W., Hunziker, E., Bosshardt, D. D., Bornstein, M. M., Seto, I., & Buser, D. (2011). The influence of PRP on early bone formation in membrane protected defects. A histological and histomorphometric study in the rabbit calvaria. Clinical Implant Dentistry and Related Research, 13(1), 1-12. doi: 10.1111/j.1708-8208.2009.00266.x

Carmona, J. (2006). Use of autologous platelet concentrates for the treatment of musculoskeletal injuries in the horse. Doctoral thesis en Medicina y Sanidad Animales, Universitat Autonoma de Barcelona. Barcelona, Espanha.

Cattaneo, M., Lecchi, A., Zighetti, M. L., & Lussana, F. (2007). Platelet aggregation studies: autologous platelet-poor plasma inhibits platelet aggregation when added to platelet-rich plasma to normalize platelet count. Haematologica, 92(5), 694-697. doi: 10.3324/haematol.10999

Chahla, J., Cinque, M. E., Piuzzi, N. S., Mannava, S., Geeslin, A. G., Murray, I. R., & LaPrade, R. F. (2017). A call for standardization in platelet-rich plasma preparation protocols and composition reporting: a systematic review of the clinical orthopaedic literature. JBJS, 99(20), 1769-1779. doi: 10.2106/JBJS.16. 01374

Choi, B. H., Zhu, S. J., Kim, B. Y., Huh, J. Y., Lee, S. H., & Jung, J. H. (2005). Effect of platelet-rich plasma (PRP) concentration on the viability and proliferation of alveolar bone cells: an in vitro study. International Journal of Oral and Maxillofacial Surgery, 34(4), 420-424. doi: 10.1016/j.ijom.2004.10. 018

Davis, V. L., Abukabda, A. B., Radio, N. M., Witt-Enderby, P. A., Clafshenkel, W. P., Cairone, J. V., & Rutkowski, J. L. (2014). Platelet-rich preparations to improve healing. Part I: workable options for every size practice. Journal of Oral Implantology, 40(4), 500-510. doi: 10.1563/AAID-JOI-D-12-00104

DeLong, J. M., Beitzel, K., Mazzocca, A. D., Shepard, D., Roller, B. L., & Hanypsiak, B. T. (2011). Update on platelet-rich plasma. Current Orthopaedic Practice, 22(6), 514-523. doi: 10.1097/BCO.0b013e318 23 bd55

DeLong, J. M., Russell, R. P., & Mazzocca, A. D. (2012). Platelet-rich plasma: the PAW classification system. Arthroscopy: The Journal of Arthroscopic & Related Surgery, 28(7), 998-1009. doi: 10.1016/j. arthro.2012.04.148

Dhurat, R., & Sukesh, M. S. (2014). Principles and methods of preparation of platelet-rich plasma: a review and author's perspective. Journal of Cutaneous and Aesthetic Surgery, 7(4), 189. doi: 10.4103/0974-2077.150734

Dragoo, J. L., Wasterlain, A. S., Braun, H. J., & Nead, K. T. (2014). Platelet-rich plasma as a treatment for patellar tendinopathy: a double-blind, randomized controlled trial. The American Journal of Sports Medicine, 42(3), 610-618. doi: 10.1177/0363546513518416

Everts, P. A., Knape, J. T., Weibrich, G., Schönberger, J. P., Hoffmann, J., Overdevest, E. P., & van Zundert, A. (2006). Platelet-rich plasma and platelet gel: a review. The Journal of Extra-Corporeal Technology, 38(2), 174. PMCID:PMC4680757

Ferrari, M., Zia, S., Valbonesi, M., Henriquet, F., Venere, G., Spagnolo, S., & Panzani, I. (1987). A new technique for hemodilution, preparation of autologous platelet-rich plasma and intraoperative blood salvage in cardiac surgery. The International Journal of Artificial Organs, 10(1), 47-50. doi: 10.1177/0 39139888701000111

Fusegawa, Y., Goto, S., Handa, S., Kawada, T., & Ando, Y. (1999). Platelet spontaneous aggregation in platelet-rich plasma is increased in habitual smokers. Thrombosis Research, 93(6), 271-278. doi: 10.10 16/s0049-3848(98)001’84-4

Giusti, I., Rughetti, A., D'Ascenzo, S., Millimaggi, D., Pavan, A., Dell'Orso, L., & Dolo, V. (2009). Identification of an optimal concentration of platelet gel for promoting angiogenesis in human endothelial cells. Transfusion, 49(4), 771-778. doi: 10.1111/j.1537-2995.2008.02033.x

Graziani, F., Ivanovski, S., Cei, S., Ducci, F., Tonetti, M., & Gabriele, M. (2006). The in vitro effect of different PRP concentrations on osteoblasts and fibroblasts. Clinical Oral Implants Research, 17(2), 212-219. doi: 10.1111/j.1600-0501.2005.01203.x

Gruber, R., Varga, F., Fischer, M. B., & Watzek, G. (2002). Platelets stimulate proliferation of bone cells: involvement of platelet derived growth factor, microparticles and membranes. Clinical Oral Implants Research, 13(5), 529-535. doi: 10.1034/j.1600-0501.2002.130513.x

Haynesworth, S. E., Kadiyala, S., Liang, L. N., & Bruder, S. P. (2002). Mitogenic stimulation of human mesenchymal stem cells by platelet release suggest a mechanism for enhancement of bone repair by platelet concentrates. Proceeding of the Meeting of the Orthopedic Research Society, Boston, MA, USA, 48.

Herter, J. M., Rossaint, J., & Zarbock, A. (2014). Platelets in inflammation and immunity. Journal of Thrombosis and Haemostasis, 12(11), 1764-1775. doi: 10.1111/jth.12730

Lima, F. L. M. (2009). Efeitos do plasma rico em plaquetas no processo de reparação de feridas dérmicas padronizadas em ratos. Tese de doutorado em Ciências Odontológicas, Universidade de São Paulo, São Paulo. doi: 10.11606/T.23.2009.tde-19122009-114243

Ljungkvist, M., Olofsson, H., Funding, E., Berntorp, E., & Zetterberg, E. (2019). Coagulation factor VIII is vital for increasing global coagulation after physical exercise. Haemophilia, 25(2), e86-e93. doi: 10.11 11/hae.13652

Malanga, G. A., & Goldin, M. (2014). PRP: review of the current evidence for musculoskeletal conditions. Current Physical Medicine and Rehabilitation Reports, 2(1), 1-15. doi: 10.1007/s40141-013-0039-5

Marx, R. E. (2001). Platelet-rich plasma (PRP): what is PRP and what is not PRP?. Implant Dentistry, 10(4), 225-228. doi: 10.1097/00008505-200110000-00002

Marx, R. E., Carlson, E. R., Eichstaedt, R. M., Schimmele, S. R., Strauss, J. E., & Georgeff, K. R. (1998). Platelet-rich plasma: growth factor enhancement for bone grafts. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology, 85(6), 638-646. doi: 10.1016/s1079-2104(98)90029-4

Michelson, A. (2007). Platelets. San Diego: Elsevier/Academic Press.

Nguyen, R. T., Borg-Stein, J., & McInnis, K. (2011). Applications of platelet-rich plasma in musculoskeletal and sports medicine: an evidence-based approach. PM&R, 3(3), 226-250. doi: 10.1016/j.pmrj.2010.11. 007

Pereira, R. C. F., Zacarias, G. V. F., Cantarelli, C., Corrêa, M. M. B., Silva, G. B. da, Barbosa, A. L. T., & De La Côrte, F. D. (2013). Avaliação de sete protocolos para obtenção de plasma rico em plaquetas na espécie equina. Ciência Rural, 43(6), 1122-1127. doi: 10.1590/S0103-84782013005000052

Peter, I., Wu, K., Diaz, R., & Borg-Stein, J. (2016). Platelet-rich plasma. Physical Medicine and Rehabilitation Clinics, 27(4), 825-853. doi: 10.1016/j.pmr.2016.06.002

Redler, L. H., Thompson, S. A., Hsu, S. H., Ahmad, C. S., & Levine, W. N. (2011). Platelet-rich plasma therapy: a systematic literature review and evidence for clinical use. The Physician and Sportsmedicine, 39(1), 42-51. doi: 10.3810/psm.2011.02.1861

Sampson, S., Gerhardt, M., & Mandelbaum, B. (2008). Platelet rich plasma injection grafts for musculoskeletal injuries: a review. Current Reviews in Musculoskeletal Medicine, 1(3-4), 165-174. doi: 10.1007/s12178-008-9032-5

Sánchez, M., Anitua, E., Azofra, J., Andía, I., Padilla, S., & Mujika, I. (2007). Comparison of surgically repaired Achilles tendon tears using platelet-rich fibrin matrices. The American Journal of Sports Medicine, 35(2), 245-251. doi: 10.1177/0363546506294078

Sánchez, M., Anitua, E., Azofra, J., Prado, R., Muruzabal, F., & Andia, I. (2010). Ligamentization of tendon grafts treated with an endogenous preparation rich in growth factors: gross morphology and histology. Arthroscopy: The Journal of Arthroscopic & Related Surgery, 26(4), 470-480. doi: 10.1016/j.arthro. 2009.08.019

Schlegel, K. A., Kloss, F. R., Schultze-Mosgau, S., Neukam, F. W., & Wiltfang, J. (2003). Osseous defect regeneration using autogenous bone alone or combined with Biogran or Algipore with and without added thrombocytes. A microradiologic evaluation. Mund-, Kiefer-und Gesichtschirurgie: MKG, 7(2), 112-118. doi: 10.1007/s10006-003-0454-3

Torricelli, P., Fini, M., Filardo, G., Tschon, M., Pischedda, M., Pacorini, A., & Giardino, R. (2011). Regenerative medicine for the treatment of musculoskeletal overuse injuries in competition horses. International Orthopaedics, 35(10), 1569-1576. doi: 10.1007/s00264-011-1237-3

Vendruscolo, C. P., Carvalho, A. D. M., Moraes, L. F., Maia, L., Queiroz, D. L., Watanabe, M. J.,... Alves, A. L. G. (2012). Avaliação da eficácia de diferentes protocolos de preparo do Plasma Rico em Plaquetas para uso em Medicina Equina. Pesquisa Veterinária Brasileira, 32(2), 106-110. doi: 10.1590/S0100-73 6X2012000200002

Weibrich, G., Hansen, T., Kleis, W., Buch, R., & Hitzler, W. E. (2004). Effect of platelet concentration in platelet-rich plasma on peri-implant bone regeneration. Bone, 34(4), 665-671. doi: 10.1016/j.bone.2003. 12.010

Woodell-May, J. E., Ridderman, D. N., Swift, M. J., & Higgins, J. (2005). Producing accurate platelet counts for platelet rich plasma: validation of a hematology analyzer and preparation techniques for counting. Journal of Craniofacial Surgery, 16(5), 749-756. doi: 10.1097/01.scs.0000180007.30115.fa

Zucker-Franklin, D. (1996). Megakaryocyte and platelet structure in thrombocytopoiesis: the effect of cytokines. Stem Cells, 14(S1), 1-17. doi: 10.1002/stem.5530140702


Semina: Ciênc. Agrár.
Londrina - PR
E-ISSN 1679-0359
DOI: 10.5433 / 1679-0359
Este obra está licenciado com uma Licença  Creative Commons Atribuição-NãoComercial 4.0 Internacional