Selective iNOS-inhibition does not influence apoptosis in ruptured canine cranial cruciate ligaments

Summary

Abnormal patterns of cell death, including increased apoptosis, can influence homeostasis of ligaments and could be involved in the pathogenesis of cranial cruciate ligament (CCL) rupture. Increased nitric oxide (NO) production has been implicated as a stimulus to increased apoptosis in articular cartilage. This study investigated apoptotic cell death in ruptured canine CCL (CCL group, n = 15), in ruptured CCL of dogs treated with oral L-N6-(1-iminoethyl)-lysine (L-NIL), a selective NO-synthetase(NOS)-inhibitor, (L-NIL group, n = 15) and compared the results with normal canine CCL (control group, n = 10).

Orally administered L-NIL at a dosage of 25 mg/m² of body surface area was effective in inhibiting NO production in the articular cartilage of dogs in the L-NIL group, but it did not significantly influence the increased quantity of apoptotic cells found in ruptured CCL specimens. The results of this study suggest that apoptosis of ligamentocytes in the canine CCL is not primarily influenced by increased NO production within the stifle joint.

Keywords

Apoptosis - caspase - cranial cruciate ligament - dog - nitric oxide

• References

• 1 Amiel D, Scotto K, Akeson WH. Cruciate ligaments, response to injury. In: Knee ligaments: structure, function, injury and repair. Daniel D, Akeson W, O’Connor J (eds). New York: Raven Press 1990; 365-377.
• 2 De Rooster H, Cox E, van Bree H. Prevalence and relevance of antibodies to type-I and -II collagen in synovial fluid of dogs with cranial cruciate ligament damage. Am J Vet Res 2000; 61: 1456-1462.
• 3 Murakami H, Shinomiya N, Kikuchi T. et al. Differential sensitivity to NO-induced apoptosis between anterior cruciate and medial collateral ligament cells. J Orthop Sci 2005; 10: 84-90.
• 4 Pelletier JP, Jovanovic DV, Lascau-Coman V. et al. Selective inhibition of inducible nitric oxide synthase reduces progression of experimental osteoarthritis in vivo: possible link with the reduction in chondrocyte apoptosis and caspase 3 level. Arthritis Rheum 2000; 43: 1290-1299.
• 5 Murakami H, Shinomiya N, Kikuchi T. et al. Up-regulated expression of inducible nitric oxide synthetase plays a key role in early apoptosis after anterior cruciate ligament injury. J Orthop Res 2006; 24: 1521-1534.
• 6 Kühn K, D’Lima DD, Hashimoto S. et al. Cell death in cartilage. Osteoarthritis Cartilage 2004; 12: 1-16.
• 7 Spreng D, Sigrist N, Jungi T. et al. Nitric oxide meta-bolite production in the cranial cruciate ligament, synovial membrane, and articular cartilage of dogs with cranial cruciate ligament rupture. Am J Vet Res 2000; 61: 530-536.
• 8 Louis E, Remer KA, Doherr MG. et al. Nitric oxide and metalloproteinases in canine articular ligaments: a comparison between the cranial cruciate, the medial genual collateral and the femoral head ligament. Vet J 2006; 172: 466-472.
• 9 Bryk R, Wolff DJ. Mechanism of inducible nitric oxide synthase inactivation by aminoguanidine and L-N6–(1–iminoethyl)lysine. Biochemistry 1998; 37: 4844-4852.
• 10 Moore WM, Webber RK, Jerome GM. et al. L-N6–(1–iminoethyl)lysine: a selective inhibitor of inducible nitric oxide synthetase. J Medicinal Chem 1994; 37: 3886-3888.
• 11 Bidlingmeyer BA, Cohen SA, Tarvin TL. Rapid analysis of amino acids using pre-column derivati -zation. J Chromatogr 1984; 336: 93-104.
• 12 Slocum B, Devine T. Tibial plateau leveling osteo -tomy for repair of cranial cruciate ligament rupture in the canine. Vet Clin North Am: Sm Anim Pract 1993; 23: 777-795.
• 13 Spreng D, Sigrist N, Schweighauser A. et al. Endogenous nitric oxide production in canine osteoarthritis: detection in urine, serum and synovial fluid specimens. Vet Surg 2001; 30: 191-199.
• 14 Granger DL, Miller WC, Hibbs Jr JB. Methods of analyzing nitric oxide production in the immune response. In: Methods in Nitric Oxide Research. Feelisch M, Stammler JS (eds). Chichester: Wiley and Sons 1996; 603-617.
• 15 Gyger O, Botteron C, Doherr M. et al. Detection and distribution of apoptotic cell death in normal and diseased canine cranial cruciate ligaments. Vet J 2007; 174: 371-377.
• 16 Muir P, Hayashi K, Manley PA. et al. Evaluation of tartrate-resistant acid phosphatase and cathepsin K in ruptured canine cranial cruciate ligament in dogs. Am J Vet Res 2002; 63: 1279-1287.
• 17 Muir P, Schamberger GM, Manley PA. et al. Locali -zation of cathepsin K and tartrate-resistant acid phospatase in synovium and cranial cruciate ligament in dogs with cruciate disease. Vet Surg 2005; 34: 239-246.
• 18 Muir P, Danova NA, Argyle DJ. et al. Collagenolytic protease expression in cranial cruciate ligament and stifle synovial fluid in dogs with cranial cruciate ligament rupture. Vet Surg 2005; 34: 482-490.
• 19 Hayashi K, Frank JD, Dubinsky C. et al. Histologic changes in ruptured canine cranial cruciate ligament. Vet Surg 2003; 32: 269-277.
• 20 Lotz M, Hashimoto S, Kühn K. Mechanisms of chondrocyte apoptosis. Osteoarthritis Cartilage 1999; 7: 389-391.
• 21 Diaz-Gallego L, Prieto JG, Coronel P. et al. Apoptosis and nitric oxide in an experimental model of osteoarthritis in rabbit after hyaluronic acid treatment. J Orthop Res 2005; 23: 1370-1376.
• 22 Pelletier JP, Jovanovic D, Fernandes JC. et al. Reduced progression of experimental osteoarthritis in vivo by selective inhibition of inducible nitric oxide synthase. Arthritis Rheum 1998; 41: 1275-1286.
• 23 Blanco FJ, Ochs RL, Schwarz H. et al. Chondrocyte apoptosis induced by nitric oxide. Am J Pathol 1995; 146: 75-85.
• 24 Kim DY, Taylor HW, Moore RM. et al. Articular chondrocyte apoptosis in equine osteoarthritis. Vet J 2003; 166: 52-57.
• 25 Kühn K, Shikhman AR, Lotz M. Role of nitric oxide, reactive oxygen species, and p38 MAP kinase in the regulation of human chondrocyte apoptosis. J Cell Physiol 2003; 197: 379-387.
• 26 Kim HA, Lee KB, Bae SC. The mechanism of low-concentration sodium nitroprusside-mediated protection of chondrocyte death. Arthritis Res Ther 2005; 7: R526-R535.
• 27 DelCarlo Jr M, Loeser RF. Nitric oxide-mediated chondrocyte cell death requires the generation of additional reactive oxygen species. Arthritis Rheum 2002; 46: 394-403.
• 28 Toda N, Toda H, Hatano Y. Nitric oxide. Involvement in the effects of anesthetic agents. Anesthesiology 2007; 107: 822-842.
• 29 Krayer M, Rytz U, Oevermann A. et al. Apoptosis of ligamentous cells of the cranial cruciate ligament from stable stifle joints of dogs with partial cranial cruciate ligament rupture. Am J Vet Res 2008; 69: 625-630.
• 30 Jauernig S, Schweighauser A, Reist M. et al. The effects of doxycycline on nitric oxide and stromelysin production in dogs with cranial cruciate ligament rupture. Vet Surg 2001; 30: 132-139.
• 31 Hayashi K, Frank JD, Hao Z. et al. Evaluation of ligament fibroblast viability in ruptured cranial cruciate ligament of dogs. Am J Vet Res 2003; 64: 1010-1016.
• 32 D’Lima DD, Hashimoto S, Chen PC. et al. Human chondrocyte apoptosis in response to mechanical injury. Osteoarthritis Cartilage 2001; 9: 712-719.
• 33 Dang AC, Warren AP, Kim HT. Beneficial effects of intra-articular caspase inhibition therapy following osteochondral injury. Osteoarthritis Cartilage 2006; 14: 526-532.