PDF herunterladen
Vet Comp Orthop Traumatol 1997; 10(01): 6-11
DOI: 10.1055/s-0038-1632561
Original Research
Schattauer GmbH

Development of an in vitro Model for the Study of the Response of Equine Tendon Fibroblasts to Injury and Medication

L. A. Dahlgren
1   From the Department of Veterinary Clinical Sciences and the College of Veterinary Medicine, Iowa State University, Ames, Iowa, USA
,
R. F. Rosenbusch
2   From the Veterinary Medical Research Institute, College of Veterinary Medicine, Iowa State University, Ames, Iowa, USA
,
L. C. Booth
1   From the Department of Veterinary Clinical Sciences and the College of Veterinary Medicine, Iowa State University, Ames, Iowa, USA
› Institutsangaben
Weitere Informationen

Publikationsverlauf

Received for publication 10. April 1996

Publikationsdatum:
22. Februar 2018 (online)

    Lizenzen und Reprints

Summary

Equine tendon fibroblasts were isolated from explants of superficial digital flexor tendon, subcultured and maintained in monolayers. The cells were characterized by light microscopy, electron microscopy and radiolabel studies for proteoglycan production. Two predominant cell morphologies were identified. The cells dedifferentiated toward a more spindle shape with repeated subcultures. Equine tendon fibroblasts were successfully cryopreserved and subsequently subcultured. The ability to produce proteoglycan was preserved.

The isolated cells were identified as fibroblasts, based on their characteristic shape by light microscopy and ultrastructure and the active production of extracellular matrix proteins. Abundant rough endoplasmic reticulum and the production of extracellular matrix products demonstrated active protein production and export. Proteoglycans were measurable via liquid scintillation counting in both the cell-associated fraction and free in the supernatant. This model is currently being utilized to study the effects of polysulfated glycosaminoglycan on tendon healing. Future uses include studying the effects of other pharmaceuticals, such as hyaluronic acid, on tendon healing.

A model was developed for in vitro investigations into tendon healing. Fibroblasts were isolated from equine superficial digital flexor tendons and maintained in monolayer culture. The tenocytes were characterized via light and electron microscopy. Proteoglycan production was measured, using radio-label techniques. The fibroblasts were cryopreserved and subsequently subcultured. The cells maintained their capacity for proteoglycan production, following repeated subculturing and cryopreservation.

Keywords

Tendinitis - cell culture - tendon fibroblast
 

  • REFERENCES

  • 1 Abrahamsson SO, Lundborg G, Lohmander LS. Recombinant human insulin-like growth factor-1 stimulates in vitro matrix synthesis and cell proliferation in rabbit flexor tendon. J Orthop Res 1991; 9: 495-502.
    CrossrefPubMedGoogle Scholar
  • 2 Becker H, Graham MF, Kelman Cohen I, Diegelmann RF. Intrinsic tendon cell proliferation in tissue culture. J Hand Surg 1981; 6: 616-9.
    CrossrefPubMedGoogle Scholar
  • 3 Birch HL. An investigation into the cellular basis of tendon degeneration (PhD Thesis). University of Bristol, Bristol, England: 1993
    Google Scholar
  • 4 Bramlage LR. Superior check ligament desmotomy as a treatment for superficial digital flexor tendonitis: initial report. Proc Annu Conv Am Assoc Equine Prac 1986; 32: 365-9.
    PubMedGoogle Scholar
  • 5 Caron JP, Eberhart SW, Nachreiner R. Influence of polysulfated glycosaminoglycan on equine articular cartilage in explant culture. Am J Vet Res 1991; 52: 1622-5.
    PubMedGoogle Scholar
  • 6 Caron JP, Toppin DS, Block JA. Effect of polysulfated glycosaminoglycan on osteoarthritic equine articular cartilage in explant culture. Am J Vet Res 1993; 54: 1116-21.
    PubMedGoogle Scholar
  • 7 Chard MD, Wright JK, Hazleman BL. Isolation and growth characteristics of adult human tendon fibroblasts. Ann Rheum Dis 1987; 46: 385-90.
    CrossrefPubMedGoogle Scholar
  • 8 Cohen IK, Diegelmann RF, Lindblad WJ. Wound Healing: Biochemical and Clinical Aspects. Philadelphia: Saunders; 1992
    Google Scholar
  • 9 Daniel JC, Mills DK. Proteoglycan synthesis by cells cultured from regions of the rabbit flexor tendon. Connect Tissue Res 1988; 17: 215-30.
    CrossrefPubMedGoogle Scholar
  • 10 Darnell J, Lodish H, Baltimore D. Molecular Cell Biology, New York: Magazine American Books; 1986
  • 11 Davis EW, Booth LC, Miller-Graber P. Effect of proximal check ligament desmotomy and exercise on healing of core lesions in the superficial digital flexor tendon of horses. Vet Surg 1994; 23: 399
    PubMedGoogle Scholar
  • 12 Fawcett DW. Bloom and Fawcett: A textbook of Histology. 12th ed.. New York: Chapman & Hall; 1994
    Google Scholar
  • 13 Ferraro GL. Lameness diagnosis and treatment in the thoroughbred racehorse. Vet Clin North Am Equine Pract 1990; 6: 63-84.
    CrossrefPubMedGoogle Scholar
  • 14 Foland JW, Trotter GW, Powers BE. et al. Effect of sodium hyaluronate in collagenaseinduced superficial digital flexor tendinitis in horses. Am J Vet Res 1992; 53: 2371-6.
    PubMedGoogle Scholar
  • 15 Freshney RI. Culture of Animal Cells. 3rd ed.. New York: Wiley-Liss; 1994
    Google Scholar
  • 16 Fries KM, Blieden T, Looney RJ. et al. Evidence of fibroblast heterogeneity and the role of fibroblast subpopulations in fibrosis. Clin Immunol Immunopathol 1994; 72: 283-92.
    CrossrefPubMedGoogle Scholar
  • 17 Fulton IC, MacLean AA, O’Reilly JL, Church S. Superior check ligament desmotomy for treatment of superficial digital flexor tendonitis in Thoroughbred and Standardbred horses. Aust Vet J 1994; 71: 233-5.
    CrossrefPubMedGoogle Scholar
  • 18 Gaughan EM, Gift LJ, DeBowes RM. et al. The influence of sequential intratendinous sodium hyaluronate on tendon healing in horses. VCOT 1995; 8: 40-5.
    PubMedGoogle Scholar
  • 19 Gelberman RH, Manske PR, Vande Berg JS. et al. Flexor tendon repair: A comparative histologic study of the rabbit, chicken, dog, and monkey. J Orthop Res 1984; 2: 39-48.
    CrossrefPubMedGoogle Scholar
  • 20 Genovese RL. Prognosis of superficial flexor tendon and suspensory ligament injuries. Proc Ann Conv Am Assoc Equine Pract 1993; 39: 17-9.
    PubMedGoogle Scholar
  • 21 Gift LJ, Gaughan EM, DeBowes RM. et al. The influence of intratendinous sodium hyaluronate on tendon healing in horses. VCOT 1992; 5: 151-7.
    PubMedGoogle Scholar
  • 22 Gillard GC, Reilly HC, Bell-Booth PG, Flint MH. The influence of mechanical forces on the glycosaminoglycan content of the rabbit flexor digitorum profundus tendon. Connect Tissue Res 1979; 7: 37-46.
    CrossrefPubMedGoogle Scholar
  • 23 Goldberg B, Rabinovitch M. Cell and Tissue Biology: A Textbook of histology. 6th ed.. Baltimore: Urban & Scherzenberg; 1988
    Google Scholar
  • 24 Goodship AE, Brown PN, Yeats JJ. et al. An assessment of filamentous carbon fibre for the treatment of tendon injury in the horse. Vet Rec 1980; 106: 217-21.
    CrossrefPubMedGoogle Scholar
  • 25 Goodship AE, Birch HL, Wilson AM. The pathobiology and repair of tendon and ligament injury. Vet Clin North Am: Equine Pract 1994; 10: 323-49.
    CrossrefPubMedGoogle Scholar
  • 26 Graham MF, Becker H, Cohen IK. et al. Intrinsic tendon fibroplasia: Documentation by in vitro studies. J Orthop Res 1984; 1: 251-6.
    PubMedGoogle Scholar
  • 27 Grant BD, Cannon HJ, Rose JA. Equine tendinitis: Results of eleven cases treated with autographs. J Equine Med Surg 1978; 2: 509-12.
    PubMedGoogle Scholar
  • 28 Hayat MA. Principles and Techniques of Electron Microscopy: Biological Applications. 3rd ed.. Boca Raton: CRC Press; 1989
    Google Scholar
  • 29 Henninger RW, Bramlage LR, Bailey M. et al. Effects of tendon splitting on experimentally- induced acute equine tendinitis. VCOT 1992; 55: 1-9.
    PubMedGoogle Scholar
  • 30 Hogan PM, Bramlage LR. Transection of the accessory ligament of the superficial digital flexor tendon for treatment of tendinitis: long term results in 61 Standardbred racehorses (1985-1992). Equine Vet J 1995; 27: 221-6.
    CrossrefPubMedGoogle Scholar
  • 31 Lindsay WK, Birch JR. The fibroblast in flexor tendon healing. Plast Reconstr Surg 1964; 34: 223-32.
    CrossrefPubMedGoogle Scholar
  • 32 Madison JB. Acute and chronic tendinitis in horses. Comp Cont Educ Pract Vet 1995; 17: 853-6.
    PubMedGoogle Scholar
  • 33 Manske PR, Gelberman RH, Vande Berg JS, Lesker PA. Intrinsic flexor-tendon repair: A morphological study in vitro. . J Bone Joint Surg 1984; 66-A: 385-96.
    PubMedGoogle Scholar
  • 34 Murphy DJ, Nixon AJ. Effects of insulin-like growth factor (IGF-1) on intrinsic metabolism of equine tendon in explant culture. Vet Surg 1994; 23: 412
    PubMedGoogle Scholar
  • 35 Pool RR. Pathology of tendons and ligaments. Equine Vet Sci 1992; 12: 145-8.
    CrossrefPubMedGoogle Scholar
  • 36 Oryan A. Experimental tendon injury and repair (PhD Thesis). University of Bristol, Bristol, England: 1989
    Google Scholar
  • 37 Redding WR, Booth LC, Pool RR. The effects of polysulfated glycosaminoglycan on the healing of collagenase induced tendonitis of the equine superficial digital flexor tendon. Vet Surg 1992; 21: 403
    PubMedGoogle Scholar
  • 38 Riederer-Henderson MA, Gauger A, Olson L. et al. Attachment and extracellular matrix differences between tendon and synovial fibroblastic cells. In Vitro 1983; 19: 127-33.
    CrossrefPubMedGoogle Scholar
  • 39 Riley CB, Bailey JV, Archer JA. Development of an in vitro explant culture model of the equine superficial digital flexor tendon. VCOT 1995; 8: 60
    PubMedGoogle Scholar
  • 40 Rooney JR, Genovese RL. A survey and analysis of bowed tendon in Thoroughbred racehorses. Equine Vet Sci 1981; 1: 49-53.
    CrossrefPubMedGoogle Scholar
  • 41 Rossdale PD, Hopes R, Digby NJW. et al. Epidemiological study of wastage among racehorses 1982 and 1983. Vet Rec 1985; 116: 66-9.
    CrossrefPubMedGoogle Scholar
  • 42 Sundqvist H, Forsskahl B, Kvist M. A promising novel therapy for achilles peritendinitis: double-blind comparison of glycosaminoglycan polysulfate and high-dose indomethacin. Int J Sports Med 1987; 8: 298-303.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 43 Taylor CM, Oelbaum RS, Grant ME. The biosynthesis of glycoproteins by cultured bovine tendon fibroblasts. Connect Tissue Res 1982; 10: 319-31.
    CrossrefPubMedGoogle Scholar
  • 44 Vogel KG, Keller EJ, Lenhoff RJ. et al. Proteoglycan synthesis by fibroblast cultures initiated from regions of adult bovine tendon subjected to different mechanical forces. Eur J Cell Biol 1986; 41: 102-12.
    PubMedGoogle Scholar
  • 45 Vogel KG, Peterson DW. Extracellular, surface, and intracellular proteoglycans produced by human embryo lung fibroblasts in culture (IMR-90). J Biol Chem 1981; 256: 13235-42.
    PubMedGoogle Scholar