Download PDF
Vet Comp Orthop Traumatol 2010; 23(06): 417-423
DOI: 10.3415/VCOT-10-03-0044
Original Research
Schattauer GmbH

Investigation of the effects of prostaglandin E2 on equine superficial digital flexor tendon fibroblasts in vitro

J. M. Cissell
1   Comparative Musculoskeletal Research Laboratory, Department of Large Animal Clinical Sciences, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, Virginia, USA
,
S. C. Milton
1   Comparative Musculoskeletal Research Laboratory, Department of Large Animal Clinical Sciences, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, Virginia, USA
,
L. A. Dahlgren
1   Comparative Musculoskeletal Research Laboratory, Department of Large Animal Clinical Sciences, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, Virginia, USA
› Author Affiliations
Further Information

Publication History

Received: 20 March 2010

Accepted: 13 June 2010

Publication Date:
19 December 2017 (online)

Also available at
    Permissions and Reprints

Summary

Objectives: To evaluate the effects of pros-taglandin E2 (PGE2) treatment on the metabolism of equine tendon fibroblasts in vitro to aid in investigating the response of tendon fibroblasts to injury and novel therapeutics.

Methods: Superficial digital flexor tendon fibroblasts isolated via collagenase digestion from six young adult horses were grown in monolayer in four concentrations of PGE2 (0, 10, 50, 100 ng/ml) for 48 hours. Cells and medium were harvested for gene expression (collagen types I and III, cartilage oligomeric matrix protein [COMP], decorin, and matrix metalloproteinase-1, –3, and –13), biochemical analysis (glycosaminoglycan, DNA, and collagen content), and cytological staining.

Results: Gene expression for collagen type I was significantly increased at 100 ng/ml PGE2 compared to 10 and 50 ng/ml. There were not any significant differences detected for gene expression of collagen type III, COMP or dec-orin or for biochemical content and cell morphology.

Clinical significance: Under the conditions investigated, exogenous treatment of equine tendon fibroblasts with PGE2 failed to alter cell metabolism in a manner useful as a model of tendon injury. A model that applies cyclic strain to a three dimensional construct seeded with tendon fibroblasts may prove to be a more useful model and merits further investigation for this purpose. The ability to assess cellular responses in an environment where the cells are supported within the extracellular matrix may prove beneficial.

Keywords

Tendon - horse - prostaglandin - fibroblast
 

  • References

  • 1 Goodship AE, Birch HL, Wilson AM. The pathobiology and repair of tendon and ligament injury. Vet Clin North Am Equine Pract 1994; 10: 323-349.
    CrossrefPubMedGoogle Scholar
  • 2 Genovese RL. Prognosis of superficial digital flexor tendon and suspensory ligament injuries. In: Proceedngs of the 39th Annual Convention of the American Association of American Equine Practitioners San Antonio, Texas, USA: 1993. December 5-8 17-19.
    Google Scholar
  • 3 Dyson S. Treatment of superficial digital flexor tendinitis: A comparison of conservative management, sodium hyaluronate, and glycosaminoglycan polysulfate. In: Proceedngs of the 39th Annual Convention of the American Association of American Equine Practitioners Texas, USA: San Antonio; 1993. December 5-8 gs. 297-300.
    Google Scholar
  • 4 Genovese RL, Reef VB, Longo KL. et al. Superficial digital flexor tendonitis: Long term sonographic and clinical study of racehorses. In: Rantanen NW, Hauser ML (eds). Proceedings of the Dubai International Equine Symposium. Bonsall, CA, USA: Matthew R. Rantanen Design; 1996. March 27-30 gs. 187-205.
    Google Scholar
  • 5 Schnabel LV, Mohammed HO, Miller BJ. et al. Platelet rich plasma (PRP) enhances anabolic gene expression patterns in flexor digitorum superficialis tendons. J Orthop Res 2007; 25: 230-240.
    CrossrefPubMedGoogle Scholar
  • 6 Waselau M, Sutter WW, Genovese RL. et al. Intralesional injection of platelet-rich plasma followed by controlled exercise for treatment of midbody suspensory ligament desmitis in Standardbred race-horses. J Am Vet Med Assoc 2008; 232: 1515-1520.
    CrossrefPubMedGoogle Scholar
  • 7 Stewart AA, Barrett JG, Byron CR. et al. Comparison of equine tendon-, muscle-, and bone marrow-derived cells cultured on tendon matrix. Am J Vet Res 2009; 70: 750-757.
    CrossrefPubMedGoogle Scholar
  • 8 Schnabel LV, Lynch ME, van der Meulen MC. et al. Mesenchymal stem cells and insulin-like growth factor-I gene-enhanced mesenchymal stem cells improve structural aspects of healing in equine flex-or digitorum superficialis tendons. J Orthop Res 2009; 27: 1392-1398.
    CrossrefPubMedGoogle Scholar
  • 9 Mitchell RD. Treatment of tendon and ligament injuries with UBM powder (ACell-Vet®). In: Proceedngs of the 39th Annual Symposium of the American College of Veterinary Surgeons Denver; Colorado, USA: 2004. October 7-9 gs. 190-193.
    Google Scholar
  • 10 Bolt DM, Ishihara A, Weisbrode SE. et al. Effects of triamcinolone acetonide, sodium hyaluronate, amikacin sulfate, and mepivacaine hydrochloride, alone and in combination, on morphology and matrix composition of lipopolysaccharide-challenged and unchallenged equine articular cartilage ex-plants. Am J Vet Res 2008. 69 861-867.
    Google Scholar
  • 11 Taintor J, Schumacher J, DeGraves F. Comparison of amikacin concentrations in normal and inflamed joints of horses following intra-articular administration. Equine Vet J 2006; 38: 189-191.
    PubMedGoogle Scholar
  • 12 Khan MH, Li Z, Wang JH. Repeated exposure of tendon to prostaglandin-E2 leads to localized tendon degeneration. Clin J Sport Med 2005; 15: 27-33.
    CrossrefPubMedGoogle Scholar
  • 13 Teitz CC, Garrett Jr WE, Miniaci A. et al. Tendon problems in athletic individuals. Instr Course Lect 1997; 46: 569-582.
    PubMedGoogle Scholar
  • 14 Flick J, Devkota A, Tsuzaki M. et al. Cyclic loading alters biomechanical properties and secretion of PGE2 and NO from tendon explants. Clin Biomech (Bristol, Avon) 2006; 21: 99-106.
    CrossrefPubMedGoogle Scholar
  • 15 Almekinders LC, Banes AJ, Ballenger CA. Effects of repetitive motion on human fibroblasts. Med Sci Sports Exerc 1993; 25: 603-607.
    PubMedGoogle Scholar
  • 16 Li Z, Yang G, Khan M. et al. Inflammatory response of human tendon fibroblasts to cyclic mechanical stretching. Am J Sports Med 2004; 32: 435-440.
    CrossrefPubMedGoogle Scholar
  • 17 Lo IK, Marchuk LL, Hollinshead R. et al. Matrix metalloproteinase and tissue inhibitor of matrix metalloproteinase mRNA levels are specifically altered in torn rotator cuff tendons. Am J Sports Med 2004; 32: 1223-1229.
    CrossrefPubMedGoogle Scholar
  • 18 Tsuzaki M, Guyton G, Garrett W. et al. IL-1 beta induces COX-2, MMP-1, –3 and –13, ADAMTS-4, IL-1 beta and IL-6 in human tendon cells. J Orthop Res 2003; 21: 256-264.
    CrossrefPubMedGoogle Scholar
  • 19 Wang JH, Jia F, Yang G. et al. Cyclic mechanical stretching of human tendon fibroblasts increases the production of prostaglandin E2 and levels of cyclooxygenase expression: a novel in vitro model study. Connect Tissue Res 2003; 44: 128-133.
    CrossrefPubMedGoogle Scholar
  • 20 Archambault JM, Wiley JP, Bray RC. Exercise loading of tendons and the development of overuse injuries. A review of current literature. Sports Med 1995; 20: 77-89.
    CrossrefPubMedGoogle Scholar
  • 21 Yang G, Im HJ, Wang JH. Repetitive mechanical stretching modulates IL-1beta induced COX-2, MMP-1 expression, and PGE2 production in human patellar tendon fibroblasts. Gene 2005; 363: 166-172.
    CrossrefPubMedGoogle Scholar
  • 22 Yamaguchi M, Shimizu N, Goseki T. et al. Effect of different magnitudes of tension force on prostaglandin E2 production by human periodontal ligament cells. Arch Oral Biol 1994; 39: 877-884.
    CrossrefPubMedGoogle Scholar
  • 23 Cilli F, Khan M, Fu F. et al. Prostaglandin E2 affects proliferation and collagen synthesis by human patellar tendon fibroblasts. Clin J Sport Med 2004; 14: 232-236.
    CrossrefPubMedGoogle Scholar
  • 24 Kuehl Jr FA, Humes JL, Ham EA. et al. Inflammation: the role of peroxidase-derived products. Adv Prostaglandin Thromboxane Res 1980; 6: 77-86.
    PubMedGoogle Scholar
  • 25 Davies P, Bailey PJ, Goldenberg MM. et al. The role of arachidonic acid oxygenation products in pain and inflammation. Annu Rev Immunol 1984; 2: 335-357.
    CrossrefPubMedGoogle Scholar
  • 26 Thampatty BP, Im HJ, Wang JH. Leukotriene B4 at low dosage negates the catabolic effect of prostaglandin E2 in human patellar tendon fibroblasts. Gene 2006; 372: 103-109.
    CrossrefPubMedGoogle Scholar
  • 27 Nixon AJ, Lust G, Vernier-Singer M. Isolation, propagation, and cryopreservation of equine articular chondrocytes. Am J Vet Res 1992; 53: 2364-2370.
    PubMedGoogle Scholar
  • 28 Farndale RW, Buttle DJ, Barrett AJ. Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. Biochim Biophys Acta 1986; 883: 173-177.
    CrossrefPubMedGoogle Scholar
  • 29 Dahlgren LA, Nixon AJ, Brower-Toland BD. Effects of beta-aminopropionitrile on equine tendon metabolism in vitro and on effects of insulin-likegrowth factor-I on matrix production by equine tenocytes. Am J Vet Res 2001; 62: 1557-1562.
    CrossrefPubMedGoogle Scholar
  • 30 Rao J, Otto WR. Fluorimetric DNA assay for cell growth estimation. Anal Biochem 1992; 207: 186-192.
    CrossrefPubMedGoogle Scholar
  • 31 Dahlgren LA, Mohammed HO, Nixon AJ. Temporal expression of growth factors and matrix molecules in healing tendon lesions. J Orthop Res 2005; 23: 84-92.
    CrossrefPubMedGoogle Scholar
  • 32 Nixon AJ, Dahlgren LA, Haupt JL. et al. Effect of adipose-derived nucleated cell fractions on tendon repair in horses with collagenase-induced tendinitis. Am J Vet Res 2008; 69: 928-937.
    CrossrefPubMedGoogle Scholar
  • 33 Leutenegger CM, Mislin CN, Sigrist B. et al. Quantitative real-time PCR for the measurement of feline cytokine mRNA. Vet Immunol Immunopathol 1999; 71: 291-305.
    CrossrefPubMedGoogle Scholar
  • 34 Arocho A, Chen B, Ladanyi M. et al. Validation of the 2–DeltaDeltaCt calculation as an alternate method of data analysis for quantitative PCR of BCR-ABL P210 transcripts. Diagn Mol Pathol 2006; 15: 56-61.
    CrossrefPubMedGoogle Scholar
  • 35 Langberg H, Boushel R, Skovgaard D. et al. Cyclooxygenase-2 mediated prostaglandin release regulates blood flow in connective tissue during mechanical loading in humans. J Physiol 2003; 551: 683-689.
    CrossrefPubMedGoogle Scholar
  • 36 Koshima H, Kondo S, Mishima S. et al. Expression of interleukin-1beta, cyclooxygenase-2, and pros-taglandin E2 in a rotator cuff tear in rabbits. J Orthop Res 2007; 25: 92-97.
    CrossrefPubMedGoogle Scholar
  • 37 Zhang J, Wang JH. Production of PGE(2) increases in tendons subjected to repetitive mechanical loading and induces differentiation of tendon stem cells into non-tenocytes. J Orthop Res 2010; 28: 198-203.
    PubMedGoogle Scholar
  • 38 Zhou D, Lee HS, Villarreal F. et al. Differential MMP-2 activity of ligament cells under mechanical stretch injury: an in vitro study on human ACL and MCL fibroblasts. J Orthop Res 2005; 23: 949-957.
    CrossrefPubMedGoogle Scholar
  • 39 Wiig ME, Amiel D, Ivarsson M. et al. Type I procollagen gene expression in normal and early healing of the medial collateral and anterior cruciate ligaments in rabbits: an in situ hybridization study. J Orthop Res 1991; 9: 374-382.
    CrossrefPubMedGoogle Scholar
  • 40 Lee J, Harwood FL, Akeson WH. et al. Growth factor expression in healing rabbit medial collateral and anterior cruciate ligaments. Iowa Orthop J 1998; 18: 19-25.
    PubMedGoogle Scholar
  • 41 Tung JT, Arnold CE, Alexander LH. et al. Evaluation of the influence of prostaglandin E2 on recombinant equine interleukin-1beta-stimulated matrix metalloproteinases 1, 3, and 13 and tissue inhibitor of matrix metalloproteinase 1 expression in equine chondrocyte cultures. Am J Vet Res 2002; 63: 987-993.
    CrossrefPubMedGoogle Scholar