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Immunohistochemical Measurement of TGF-β1 and Factor VIII in the Skin of Horses Treated with Leukocyte-poor Platelet-rich Plasma: A Randomized Controlled Trial

1Departamento de Veterinária, Universidade Federal de Viçosa, Viçosa, Minas Gerais, Brazil
2Laboratório de Processamento de Tecido, Universidade Federal de São João del-Rei, Divinópolis, MG, Brazil
3Laboratório de Patologia Experimental, Universidade Federal de São João del-Rei, Divinópolis, MG, Brazil

Special Issues: Regenerative Medicine: Hot Topics and Developing Trends

Platelet-rich plasma (PRP) is a treatment used for several diseases of various species and different body regions. However, there is controversy about its therapeutic efficacy. The objective of this experiment was to immunohistochemically evaluate the expression of transforming growth factor beta 1 (TGF-β1) and factor VIII during the healing period of skin treated with leukocyte-poor platelet-rich plasma (LP-PRP) in seven healthy gelding crossbred horses. This study was a randomized controlled clinical trial. Wounds were surgically induced in gluteal region (treated group), and LP-PRP (0.5 mL) was injected into all borders of the wounds 12 h later. The contralateral region (untreated) was adopted as the control group. Five skin samples were obtained from the treated and untreated groups for immunohistochemical analysis. The last one was collected upon wound closure. We hypothesized that the treated wounds would exhibit higher expression of both TGF-β1 and factor VIII than the untreated wounds. There was no difference between groups in TGF-β1 immunostaining. However, a difference was observed between groups in immunostaining of blood vessels for factor VIII. This difference (P < 0.05) occurred after macroscopic wound closure, with larger numbers of vessels stained in the treated wounds. The administration of a single dose of LP-PRP 12 h after wound induction in horses is not effective compared with the physiological healing of horse skin. Further research should not only take into consideration the association of the platelet-rich component with stem cells, which, in general, have provided adequate responses, but also evaluate other proteins present in PRP besides those typically studied.
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Keywords equine; growth factors; platelet-rich concentrate; wound; transforming growth factor beta 1; von Willebrand

Citation: Maria V. Souza, Mariana B. Silva, Hélio B. Santos, Rosy I. M. A. Ribeiro, Thayne O. Silva, Ralph G. Thomé. Immunohistochemical Measurement of TGF-β1 and Factor VIII in the Skin of Horses Treated with Leukocyte-poor Platelet-rich Plasma: A Randomized Controlled Trial. AIMS Cell and Tissue Engineering, 2017, 1(2): 84-103. doi: 10.3934/celltissue.2017.2.84


  • 1. Sanchez-Casanova RE, Masri-Daba M, Alonso-Díaz MÁ, et al. (2014) Prevalence of cutaneous pathological condition and factors associated with the presence of skin wounds in working equids in tropical regions of Veracruz, Mexico. Trop Anim Health Prod 46: 555-561.    
  • 2. Pessoa AFA, Pessoa CRM, Neto EGM, et al. (2014) Doenças de pele em equídeos no semiárido brasileiro. Pesq Vet Bras 34: 743-748.    
  • 3. Enoch S, Leaper DJ (2007) Basic science of wound healing. Surgery 26: 31-37.
  • 4. Harper D. Young A, McNaugh C-E (2011) The physiology of wound healing. Surgery 32: 445-450.
  • 5. Schultz GS, Ladwig G, Wysocki A (2005) Extracellular matrix: review of its roles in acute and chronic wounds. World Wide Wounds 1-12.
  • 6. Mei-Dan O, Lippi G, Sánchez M, et al. (2010) Autologous platelet-rich plasma: a revolution in soft tissue sports injury management? Phys Sportsmed 38: 127-135.    
  • 7. Sutter WW, Kaneps AJ, Bertone AL, et al. (2004) 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 65: 924-930.
  • 8. Dhurat R, Sukesh MS (2014) Principles and methods of preparation of platelet rich plasma: a review and author's perpective. J Cutan Aesthet Surg 4: 189-197.
  • 9. Fitzpatrick J, Bulsara MK, McCrory PR, et al. (2017) Analysis of platelet-Rich plasma extraction. Variations in platelet and blood components 4 common commercial kits. Orthop J Sport Med 5: 1-8.
  • 10. Argüelles D, Carmona JU, Climent F, et al. (2008) Autologous platelet concentrates as a treatment for musculoskeletal lesions in 5 horses. Vet Rec 162: 208-211.
  • 11. Engebretsen L, Steffen K, Alsousou J, et al. (2010) IOC consensus paper on the use of platelet-rich plasma in sports medicine. Br J Sports Med 44: 1072-1081.    
  • 12. Zandim BM, Souza MV, Frassy L, et al. (2013) Immunohistochemistry of fator VIII, histology and morphometry in equine tendon treated with platelet-rich plasma. Rev Bras Med Vet 35: 169-184.
  • 13. Moraes APL, Moreira JJ, Brossi PM, et al. (2015) Short- and long-term effects of platelet-rich plasma upon healthy equine joints: Clinical and laboratory aspects. Can Vet journal La Rev Vétérinaire Can 56: 831-838.
  • 14. Bembo F, Eraud J, Philandrianos C, et al. (2016) Combined use of platelet rich plasma & micro-fat in sport and race horses with degenerative joint disease: preliminary clinical study in eight horses. Muscles Ligaments Tendons J 6: 198-204
  • 15. Yümün G, Kahraman C, Kahraman N, et al. (2016) Effects of hyperbaric oxygen therapy combined with platelet-rich plasma on diabetic wounds: an experimental rat model. Arch Med Sci 12: 1370-1376.
  • 16. Anitua E, Alkhraisat MH, Orive G (2012) Perspectives and challenges in regenerative medicine using plasma rich in growth factors. J Control Release 157: 29-38.
  • 17. Boswell SG, Schnabel LV, Mohammed OH (2014) Increasing platelet concentrations in leukocyte-reduced platelet-rich plasma decrease collagen gene synthesis in tendons. Am J Sports Med 42: 42-49.
  • 18. Souza MV, Pinto JO, Costa MBM, et al. (2014) Expressão gênica do colágeno em ferida cutânea de equinos tratada com plasma rico em plaquetas. Pesq Vet Bras 34: 233-240.    
  • 19. Ehrenfest DM, Andia I, Zumstein MA, et al. (2014) Classification of platelet concentrates (platelet-rich plasma-PRP, platelet-rich fibrin-PRF) for topical and infiltrative use in orthopedic and sports medicine: current consensus, clinical implications and perspectives. Muscles Ligaments Tendons J 8: 3-9.
  • 20. Dragoo JL, Braun HJ, Durham JL, et al. (2012) Comparison of the acute inflammatory response of two commercial platelet-rich plasma systems in healthy rabbit tendons. Am J Sports Med 40: 1274-1281.    
  • 21. Fitzpatrick J, Bulsara M, Zheng MF (2016) The effectiveness of platelet-rich plasma in the treatment of tendinopathy. A meta-analysis of randomized controlled clinical trials. Am J Sports Med 45: 226-233.
  • 22. Jubert NJ, Rodriguez L, Reverté-Vinaixa MM, et al. (2017) Platelet-rich plasma injections for advanced knee osteoarthritis: a prospective, randomized, bouble-blinded clinical trial. Orthop J Sports Med 5. In Press.
  • 23. Bennell KL, Hunter DJ, Paterson KL (2017) Platelet-rich plasma for the management of hip and knee osteoarthritis. Curr Rheumatol Rep 24: 1-10.
  • 24. Souza MV, Silva MB, Pinto JO, et al. (2015) Immunohistochemical expression of collagens in the skin of horses treated with leukocyte-poor platelet-rich plasma. BioMed Res Internat 1-12.
  • 25. Kraeutler MJ, Garabekyan T, Mei-Dan O (2017) The use of platelet-rich plasma to augment conservative and surgical treatment of hip and pelvic disorders. Muscles Ligaments Tendons J 6: 409-419.
  • 26. Bhushan M, Young HS, Brenchley PE, et al. (2002) Recent advances in cutaneous angiogenesis. Br J Dermatol 147: 418-425.
  • 27. Theoret CL, Wilmink JM (2013) Aberrant wound healing in the horse: naturally occurring conditions reminiscent of those observed in man. Wound Repair Regen 2: 365-371.
  • 28. Pufe T, Petersen WJ, Mentlein R, et al. (2005) The role of vasculature and angiogenesis for the pathogenesis of degenerative tendons disease. Scand J Med Sci Sports 15: 211-222.
  • 29. Theoret CL, Barber SM, Moyana TN (2002) Preliminary observations on expression of transforming growth factors β1 and β3 in equine full-thickness skin wounds healing normally or with exuberant granulation tissue. Vet Surg 31: 266-773.    
  • 30. van den Boom R, Wilmink JM, O'Kane S, et al. (2002) Transforming growth factor-b levels during second intention healing are related to the different course of wound contraction in horses and ponies. Wound Repair Regen 10: 188-194.    
  • 31. Eppley BL, Woodel JE, Higgins J (2004) Platelet quantification and growth factor analysis from platelet-rich plasma: implications for wound healing. Plastic Reconstr Surg 114: 1502-1508.
  • 32. Bosch G, Moleman M, Barneveld A, et al. (2011) The effect of platelet-rich plasma on the neovascularization of surgically created equine superficial digital flexor tendon lesions. Scand J Med Sci Sport 21: 554-561.
  • 33. Rosenberg JB, Greengard JS, Montgomery RR (2000) Genetic induction of a releasable pool of factor VIII in human endothelial cells. Arterioscler Thromb Vasc Biol 20: 2689-2695.    
  • 34. DeRossi R, Coelho AC, Mello GS, et al. (2009) Effects of platelet-rich plasma gel on skin healing in surgical wound in horses. Acta Cir Bras 24: 276-281.    
  • 35. Monteiro SO, Lepage OM, Theoret CL (2009) Effects of platelet-rich plasma on the repair of wounds on the distal aspect of the forelimb in horses. Am J Vet Res 70: 277-282.    
  • 36. Souza MV, Pinto JO, Costa MM, et al. (2014) Quantificação de fatores de crescimento na pele de equinos tratada com plasma rico em plaquetas. Pesq Vet Bras 34: 599-612.
  • 37. Deschene K, Celeste C, Boerboom D, et al. (2011) Constitutive expression of hypoxia-inducible factor-1 a in keratinocytes during the repair of skin wounds in horses. Wound Repair Regen 19: 250-259.    
  • 38. Theoret CL, Barber SM, Gordon JR (2002) Temporal localization of immunoreactive transforming growth factor β1 in normal equine skin and in full-thickness dermal wounds. Vet Surg 31: 274-280.
  • 39. Borena BM, Martens A, Broeckx SY, et al. (2015) Regenerative skin wound healing in mammals: state-of-the-art on growth factor and stem cell based treatments. Cell Physiol Biochem 36: 1-23.    
  • 40. Pruna R (2016) Influence of single nucleotide polymorphisms on the ffficacy of platelet-rich plasma in the treatment of injuries. J Ost Arth 1: 1-2.
  • 41. Hessel LN, Bosch G, van Weeren, et al. (2015) Equine autologous platelet concentrates: a comparative study between different available systems. Equine Vet J 47: 319-325.    
  • 42. Maia L, Souza MV, Júnior JIR, et al. (2009) Platelet-rich plasma in the treatment of induced tendinopathy in horses: histologic evaluation. J Equine Vet Sci 29: 618-626.
  • 43. Anitua E, Andia I, Sanchez M, et al. (2004) Autologous platelet source of proteins for healing and tissue regeneration. J Thromb Haemost 91: 4-15.
  • 44. Graziani F, Ivanovski S, Cei S, et al. (2006) The in vitro effect of different PRP concentrations on osteoblasts and fibroblasts. Clin Oral Implants Res 17: 212-219.    
  • 45. Andia I, Sánchez M, Maffulli N (2012) Joint pathology and platelet-rich plasma therapies. Expert Opin Biol Ther 12: 7-22.    
  • 46. Farghali HA, AbdEIKader NA, Khattab MS, et al. (2017) Evaluation of subcutaneous infiltration of autologous platelet-rich plasma on skin-wound healing in dogs. Biosci Rep 37: 1-13.
  • 47. Karayannopoulou M, Psalla D, Kazakos G, et al. (2015) Effect of locally injected autologous platelet-rich plasma on second intention wound healing of acute full-thickness skin defects in dogs. Vet Comp Orthop Traumatol 28: 172-178.
  • 48. Marques MEM, Laposy CB, Silva MLS, et al. (2017) Collagen quantification in rabbit dermal wound treated with heterologous platelet-rich plasma gel. Semina: Cienc Agrar 38: 249-258.
  • 49. Patruno M, Martinello T (2014) Treatments of the injured tendon in veterinary medicine: From scaffolds to adult stem cells. Histol Histopathol 29: 417-422.
  • 50. Fisher MB, Mauck RL (2013) Tissue engineering and regenerative medicine: recent innovations and the transition to translation. Tissue Eng Part B 19: 1-13.
  • 51. Morgan DD, McClure S, Yaeger MJ, et al. (2009) Effects of extracorporeal shock wave therapy on wounds of the distal portion of the limbs in horses. J Am Vet Med Assoc 234: 1154-1161.    
  • 52. Schwartz AJ, Wilson DA, Keegan KG, et al. (2002) Factors regulating collagen synthesis and degradation during second-intention healing of wounds in the thoracic region and the distal aspect of the forelimb of horses. Am J Vet Res 63: 1564-1570.
  • 53. Beldon P (2010) Basic science of wound healing. Surgery (Oxford) 28: 409-412.    
  • 54. Schultz GS, Davidson JM, Kirsner RS, et al. (2011) Dynamic reciprocity in the wound microenvironment. Wound Repair Regen 19: 134-148.
  • 55. Yamauchi M, Mechanic GL (1988) Cross-linking of collagen. In: NIMNI, M. E. Collagen: Biochemistry, Biomechanics, Biotechnology. Boca Raton, Flórida: C.R.C. Press, 1: 157-172.
  • 56. Theoret C (2009) Tissue engineering in wound repair: the three 'R's--repair, replace, regenerate. Vet Surg 38: 905-913.    
  • 57. O'Kane S, Ferguson MWJ (1997) Transforming growth factor βs and wound healing. Int J Biochem Cell Biol 29: 63-78.
  • 58. Broeckx S, Maes S, Martinello T, et al. (2014) Equine epidermis: a source of epithelial-like stem/progenitor cells with in vitro and in vivo regenerative capacities. Stem Cells Dev 23: 1134-1148.
  • 59. Roubelakis MG, Trohatou O, Roubelakis A, et al. (2014) Platelet-rich plasma (PRP) promotes fetal mesenchymal stem/stromal cell migration and wound healing process. Stem Cell Rev 10: 417-428.    
  • 60. Baksh N, Hannon CP, Murawski CD, et al. (2013) Platelet-rich plasma in tendon models: a systematic review of basic science literature. Artrosc J Arthrosc Rel Surg 29: 596-607.    
  • 61. Brossi PM, Moreira JJ, Machado TS, et al. (2015) Platelet-rich plasma in orthopedic therapy: a comparative systematic review of clinical and experimental data in equine and human musculoskeletal lesions. BMC Vet Res 11: 1-17.    
  • 62. Grambart ST (2015) Sports medicine and platelet-rich plasma: nonsurgical therapy. Clin Podiatr Med Surg 32: 99-107.    
  • 63. Witte S, Dedman C, Harriss F, et al. (2016) Comparison of treatment outcomes for superficial digital flexor tendonitis in National Hunt racehorses. Vet J 216: d157-163.    
  • 64. Lynch MD, Bashir S (2016) Applications of platelet-rich plasma in dermatology: a critical appraisal of the literature. J Dermatolog Treat 1-5.
  • 65. Fraria R, Berta L (2011) Biological effects of extracorporeal shock waves on fibroblasts. A review. Muscles Ligaments Tendons J 1: 138-147.
  • 66. Wasterlain AS, Braun HJ, Dragoo JL (2012) Contents and formulations of platelet-rich plasma. Oper Tech Orthop 22: 33-42.    
  • 67. Boswell SG, Cole BJ, Sundman EA, et al. (2012) Platelet-rich plasma: a milieu of bioactive factors. Arthroscopy 28: 429-439.


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Copyright Info: 2017, Maria V. Souza, et al., licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution Licese (http://creativecommons.org/licenses/by/4.0)

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