Download PDF
Vet Comp Orthop Traumatol 2006; 19(01): 43-47
DOI: 10.1055/s-0038-1632972
Original Research
Schattauer GmbH

Anabolic effects of acellular bone marrow, platelet rich plasma, and serum on equine suspensory ligament fibroblasts in vitro

J. J. Smith
1   Department of Veterinary Clinical Studies, University of Pennsylvania, New Bolton Center, 382 West Street Rd, Kennett Square, Pennsylvania, USA
,
M. W. Ross
1   Department of Veterinary Clinical Studies, University of Pennsylvania, New Bolton Center, 382 West Street Rd, Kennett Square, Pennsylvania, USA
,
R. K. W. Smith
2   Department of Clinical Sciences, Royal Veterinary College, North Mymms, Hertfordshire, UK
› Author Affiliations
Further Information

Publication History

Received 10 March 2005

Accepted 05 July 2005

Publication Date:
08 February 2018 (online)

    Permissions and Reprints

Summary

The purpose of this study was to investigate the response of suspensory ligament fibroblasts (SLF) to in vitro stimulation using acellular bone marrow (ABM), platelet rich plasma (in vitro PRP), and serum as potential treatment modalities for suspensory desmitis. Blood, bone marrow, and suspensory ligaments were collected from five horses. SLF were harvested, grown until confluent, and stimulated with various concentrations of ABM, PRP, equine serum, foetal bovine serum, and medium (control). The responses to the treatments were assessed using a combination of radiolabeling for total protein synthesis and an ELISA for quantification of Cartilage Oligomeric Matrix Protein (COMP) production. Addition of all of the samples resulted in significant increases in COMP and total protein synthesis over controls (P<0.001). ABM caused the greatest increase in both COMP and total protein synthesis by the SLF. Equine ABM, PRP, and serum contain anabolic factors that promote matrix synthesis by SLF in vitro, with ABM having the greatest effect. Application of bone marrow to injured ligaments may enhance healing by providing anabolic factors, other than or in addition to mesenchymal stem cells, which stimulate matrix production.

Keywords

Bone marrow aspirate - platelet rich plasma - serum - growth factor - suspensory desmitis.
 

  • References

  • 1 Aspenberg P, Forslund C. Enhanced healing with GDF 5 and 6. Acta Orthop Scand 1999; 70: 51-4.
    CrossrefPubMedGoogle Scholar
  • 2 Bennett NT, Schultz GS. Growth factors and wound healing: biochemical properties of growth factors and their receptors. Am J Surg 1993; 165: 728-36.
    CrossrefPubMedGoogle Scholar
  • 3 Carter CA, Jolly DG, Worden CE. Platelet rich plasma gel promotes differentiation and regeneration during equine wound healing. Exp Mol Pathol 2003; 74: 244-55.
    CrossrefPubMedGoogle Scholar
  • 4 Chan BP, Fu S, Qin L. Effects of basic fibroblast growth factor on early stages of tendon healing. Acta Orthop Scand 2000; 71: 513-8.
    CrossrefPubMedGoogle Scholar
  • 5 Clemmons RM, Bliss EL, Dorsey-Lee MR. Platelet function, size, and yield in whole blood and in platelet-rich-plasma prepared using differing centrifugation force and time in domestic and food-producing animals. Thromb Haemost 1983; 50: 838-43.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 6 Dahlgren LA, Rosenbusch RF, Booth LC. Development ofanin vitro model for the study of the response of equine tendon fibroblasts to injury and medication. Vet Comp Orthop Traumatol 1997; 10: 6-11.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 7 Dahlgren LA, Nixon AJ, Brower-Toland BD. Effects of ß-aminoproprionitrile on equine tendon metabolism in vitro and on effects of insulin-like growth factor-I on matrix protein production by equine tenocytes. Am J Vet Res 2001; 62: 1557-62.
    CrossrefPubMedGoogle Scholar
  • 8 Dahlgren LA, van der Meulen. MCH, Bertram JEA. Insulin-like growth factor-I improves cellular and molecular aspects ofhealing in a collagenase-induced model of flexor tendonitis. J Orthop Res 2002; 20: 910-9.
    PubMedGoogle Scholar
  • 9 DesRosiers EA, Yahia L, Rivard CH. Proliferative and matrix synthesis response of canine anterior cruciate ligament fibroblasts submitted to combined growth factors. J Orthop Res 1996; 14: 200-8.
    CrossrefPubMedGoogle Scholar
  • 10 Dyson SJ, Genovese RL. The suspensory apparatus. In: Lameness in Horses. Ross MW, Dyson SJ (eds). Philadelphia: Saunders 2003; 654-66.
    PubMedGoogle Scholar
  • 11 Elliott J, Berhane Y, Bailey SR. Effects of monoamines formed in the cecum of horses on equine digital blood vessels and platelets. Am J Vet Res 2003; 64: 1124-31.
    CrossrefPubMedGoogle Scholar
  • 12 Evans CH. Cytokines and the role they play in the healing of tendons and ligaments. Sports Med 1999; 28: 71-6.
    CrossrefPubMedGoogle Scholar
  • 13 Herthel DJ. Enhanced suspensory ligament healing in 100 horses by stem cell and other bone marrow components. Proc Am Assoc Equine Pract 2001; 47: 319-21.
    PubMedGoogle Scholar
  • 14 Kang HJ, Kang ES. Ideal concentration of growth factors in rabbit's flexor tendon culture. Yonsei Med J 1999; 40: 26-9.
    CrossrefPubMedGoogle Scholar
  • 15 Kingston JK, Bayly WM, Sellon DC. Effects of sodium citrate, low molecular weight heparin, and prostaglandin E1 on aggregation, fibrinogen binding, and enumeration of platelets. Am J Vet Res 2001; 62: 547-54.
    CrossrefPubMedGoogle Scholar
  • 16 Letson AK, Dahners LE. The effect of combinations of growth factors on ligament healing. Clin Orthop 1994; 308: 207-12.
    PubMedGoogle Scholar
  • 17 Marui T, Niyibizi C, Georgescu HI. Effect of growth factors on matrix synthesis by ligament fibroblasts. J Orthop Res 1997; 15: 18-22.
    CrossrefPubMedGoogle Scholar
  • 18 Marx RE. Platelet-Rich Plasma: A source of multiple autologous growth factors for bone grafts. In: Tissue Engineering: Application in Maxillofacial Surgery and Periodontics. Lynch SE, Genco RJ, Marx RE (eds). Chicago: Quintessence 1999; 71-82.
    PubMedGoogle Scholar
  • 19 Munn DH, Shafizadeh E, Attwood JT. Inhibition of T cell proliferation by macrophage tryptophan catabolism. J Exp Med 1999; 189: 1363-72.
    CrossrefPubMedGoogle Scholar
  • 20 Murphy DJ, Nixon AJ. Biochemical and site-specific effects of insulin-like growth factor-I on intrinsic tenocyte activity in equine flexor tendons. Am JVet Res 1997; 58: 103-9.
    PubMedGoogle Scholar
  • 21 Smith RKW, Zunino L, Webbon PM. The Distribution of Cartilage Oligomeric Matrix Protein (COMP) in tendon and its variation with tendon site, age and load. Matrix Biol 1997; 16: 255-71.
    CrossrefPubMedGoogle Scholar
  • 22 Smith RKW, Birch HL, Goodman S. The influence of ageing and exercise on tendon growth and degeneration-hypotheses for the initiation and prevention of strain-induced tendinopathies. Comp Biochem Physiol A Mol Integr Physiol 2002; 13: 1039-50.
    PubMedGoogle Scholar
  • 23 Smith RKW, Gerard M, Dowling B. Correlation of Cartilage Oligomeric Matrix Protein (COMP) levels in equine tendon with mechanical properties – a proposed role for COMP in determining function-specific mechanical characteristics of locomotor tendons. Equine Vet J Suppl 2002; 34: 241-4.
    PubMedGoogle Scholar
  • 24 Smith RKW. Pathophysiology of tendon injury. In: Lameness in Horses. Ross MW, Dyson SJ. (eds) Philadelphia: Saunders; 2003: 616-26.
    Google Scholar
  • 25 Smith RKW, Korda M, Blunn GW. Isolation and implication of autologous equine mesenchymal stem cells from bone marrow into the superficial digital flexor tendon as a potential novel treatment. Equine Vet J 2003; 35: 99-102.
    PubMedGoogle Scholar
  • 26 Spindler KP, Imro AK, Mayes CE. Patellar tendon and anterior cruciate ligament have different mitogenic responses to platelet-derived growth factor and transforming growth factor-fi. J Orthop Res 1996; 14: 542-6.
    CrossrefPubMedGoogle Scholar
  • 27 Woo SL, Hildebrand K, Watanabe N. Tissue engineering of ligament and tendon healing. Clin Orthop 1999; 367S: S312-S323.
    PubMedGoogle Scholar