Biochemical and site-specific effects of insulin-like growth factor I on intrinsic tenocyte activity in equine flexor tendons.
OBJECTIVE
To examine the site-specific and dose-dependent effects of insulin-like growth factor I (IGF-I) on normal equine tendon in vitro.


METHODS
Superficial digital flexor tendon explants derived from a euthanatized 3-year-old horse.


METHODS
Explants in culture were treated with 0, 100, 250, or 500 ng of IGF-I/ml for 14 days with an end-stage radiolabel of 20 microCi of [3H]proline/ml or 5 microCi of [3H]thymidine/ml. The tendon tissues were then analyzed biochemically for hydroxyproline content by reverse-phase high-performance liquid chromatography, DNA content by fluorometry, and glycosaminoglycan content by the dimethylmethylene blue dye-binding assay. In addition, morphologic analysis of the explants comprised histologic examination, autoradiography, and immunohistochemistry.


RESULTS
Hydroxyproline content was significantly increased in explants treated with 100 and 250 ng of IGF-I/ml. Additionally, the collagen synthetic rate, measured by incorporation of [3H]proline into hydroxyproline, was significantly increased for all treatment groups. On the basis of autoradiograms, fibroblast proliferation and collagen synthesis were predominantly confined to the endcap and adjacent endotenon of the explants. Enhanced immunoreactivity for type-I collagen, compared with type-III collagen, was evident in the treated explants, an observation supported by positive staining for type-I collagen with picrosirius red. Histologically, treated explants contained greater numbers of larger and more metabolically active fibroblasts, compared with untreated controls.


CONCLUSIONS
IGF-I enhances collagen synthesis in normal equine flexor tendon in a dose-dependent manner. IGF-I also exerts its primary effect on cell proliferation and collagen synthesis in the epitenon and adjacent endotenon and accompanying perivascular connective tissues, consistent with enhancement of intrinsic tendon metabolism.


CONCLUSIONS
IGF-I may have a potential role in the treatment of tendinitis in horses.
Murphy
D J
DJ
Comparative Orthopaedics Laboratory, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
Nixon
A J
AJ
D000818:Animals; D001345:Autoradiography / Q000379:methods / Q000662:veterinary; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002851:Chromatography, High Pressure Liquid / Q000379:methods / Q000662:veterinary; D003094:Collagen / Q000378:metabolism; D004247:DNA / Q000032:analysis / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D006025:Glycosaminoglycans / Q000032:analysis / Q000378:metabolism; D006736:Horses / Q000378:metabolism* / Q000502:physiology; D006909:Hydroxyproline / Q000032:analysis / Q000378:metabolism; D007150:Immunohistochemistry; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology*; D009924:Organ Culture Techniques; D011392:Proline / Q000032:analysis / Q000378:metabolism; D013710:Tendons / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D013936:Thymidine / Q000032:analysis / Q000378:metabolism; D014316:Tritium
D006025:Glycosaminoglycans; D014316:Tritium; D007334:Insulin-Like Growth Factor I; D003094:Collagen; D004247:DNA; D011392:Proline; D006909:Hydroxyproline; D013936:Thymidine