Veterinary Research Communications

, Volume 32, Issue 3, pp 243–253 | Cite as

The effects of enrofloxacin on canine tendon cells and chondrocytes proliferation in vitro

  • S. Lim
  • M. A. Hossain
  • J. Park
  • S. H. Choi
  • G. KimEmail author
Original Article


Enrofloxacin, a fluoroquinolone antibiotic has been used widely in humans and domestic animals, including dogs, because of its broad-spectrum activity and relative safety. The side effects of fluoroquinolone, induced tendinopathy, tendonitis, spontaneous tendon rupture and cartilage damage, remain incompletely understood. In the present study, we investigated the in vitro effects of enrofloxacin on cell proliferation and induction of apoptosis in canine Achilles tendon cells and chondrocytes. Cell growth and proliferation after treating with enrofloxacin for 2–6 days was quantified by a colorimetric 2,3-bis{2-methoxy-4-nitro-5-sulfophenyl}-2H-tetrazolium-5-carboxyanilide inner salt (XTT) assay. The results showed that enrofloxacin could inhibit the proliferation of canine tendon cells and chondrocytes at increasing concentrations (10–200 μg/ml). The inhibition of proliferation of canine tendon cells and chondrocytes after exposure to enrofloxacin were associated with induction of apoptosis, as evidenced by the typical nuclear apoptotic condensed nuclei found using Hoechst 33258 staining. It was demonstrated that canine tendon cells and chondrocytes treated with 200 μg/ml enrofloxacin for 4 days exhibited apoptotic features and fragmentation of DNA. Enrofloxacin also increased the apoptosis of canine tendon cells and chondrocytes in a dose and time-dependent manner. The results indicate that enrofloxacin inhibits cell proliferation, induces apoptosis and DNA fragmentation, which might explain enrofloxacin-induced tendinopathy and cartilage damage.


Apoptosis Canine tendon cells Cell proliferation Chondrocytes Enrofloxacin 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Beluche, L. A., Bertone, A. L., Anderson, D. E., Kohn, C. W. and Weisbrode, S. E., 1999. In vitro dose-dependent effects of enrofloxacin on equine articular cartilage. American Journal of Veterinary Research, 60, 577-583.PubMedGoogle Scholar
  2. Corps, A. N., Harrall, R. L., Curry, V. A., Fenwick, S. A., Hazleman, B. L and Riley, G. P., 2002. Ciprofloxacin enhances the stimulation of Matrix metalloproteinase 3 expression by interleukin-1beta in human tendon-derived cells. A potential mechanism of fluoroquinolone-induced tendinopathy. Arthritis and Rheumatism, 46, 3034-3040.PubMedCrossRefGoogle Scholar
  3. Davenport, C. L., Boston, R. C. and Richardson, D. W., 2001. Effects of enrofloxacin and magnesium deficiency on matrix metabolism in equine articular cartilage. American Journal of Veterinary Research, 62,160-166.PubMedCrossRefGoogle Scholar
  4. Eastman, A. and Barry, M. A., 1992. The origins of DNA breaks: a consequesnce of DNA damage, DNA repair or apoptosis? Cancer Investigation, 10, 229-240.PubMedCrossRefGoogle Scholar
  5. Egerbacher, M., Edinger, J. and Tschulenk, W., 2001. Effects of enrofloxacin and ciprofloxacin hydrochloride on canine and equine chondrocytes in culture. American Journal of Veterinary Research, 62, 704-708.PubMedCrossRefGoogle Scholar
  6. Greene, C. E. and Budsberg, S. G., 1993. Veterinary use of quinolones. In Hooper D. C. and Wolfson, J. S. (eds): Quinolone antimicrobial agents, 2nd edn. pp. 473-488. Washington, DC: American Society for Microbiology, 1993.Google Scholar
  7. Haddow, L. J., Chandra Sekhar, M., Hajela, V. and Gopal Rao, G., 2003. Spontaneous Achilles tendon rupture in patients treated with levofloxacin. Journal of Antimicrobial Chemotherapy (London), 51, 747-748.CrossRefGoogle Scholar
  8. Hunziker, E. B., 1999. Biologic repair of articular cartilage. Defect models in experimental animals and matrix requirements. Clinical Orthopaedics and related Research, 367, 135-146.CrossRefGoogle Scholar
  9. Kashida, Y. and Kato, M., 1997. Characterization of fluoroquinolone-induced Achilles tendon toxicity in rats: comparism of toxicities of 10 fluoroquinolones and the effects of anti-inflammatory compounds. Antimicrobial Agents Chemotherapy, 41, 2389-2393.Google Scholar
  10. Kato, M., Takada, S., Kashida, Y. and Nomura, M., 1995. Histological examination on Achilles tendon lesions induced by quinolone antibacterial agents in juvenile rats. Toxicologic Pathology, 23, 385-392.PubMedCrossRefGoogle Scholar
  11. Khaliq, Y. and Zhanel, G. G., 2003. Fluoroquinolone-associated tendinopathy: a critical review of the literature. Clinical Infectious Diseases, 36, 1404-1410.PubMedCrossRefGoogle Scholar
  12. Le Huec, J. C., Schaeverbeke, T., Chauveaux, D., Rivel, J., Dehais, J. and LeRebeller, A. 1995. Epicondylitis after treatment with fluoroquinolone antibiotics. Journal of Bone Joint Surgery. British (London), 77, 293-295.Google Scholar
  13. Lewis, J. R., Gums, J. G. and Dickensheets, D. L., 1999. Levofloxacin-induced bilateral Achilles tendonitis. Annals of Pharmacotherapy, 33, 792-795.PubMedCrossRefGoogle Scholar
  14. Lipsky, B. A. and Baker, C. A., 1999. Fluoroquinolone toxicity profiles: a review focusing on newer agents. Clinical Infectious Diseases, 28, 352-364.PubMedCrossRefGoogle Scholar
  15. O’Driscoll, S. W., 1998. The healing and regeneration of articular cartilage. Journal of Bone Joint Surgery. America (Boston MA), 80, 1795-1812.Google Scholar
  16. Owens, R. C. Jr. and Ambrose, P. G., 2005. Antimicrocrobial safety: focus on fluoroquinolones: focus on fluoroquinolones. Clinical Infectious Diseases, 15, 144-157.CrossRefGoogle Scholar
  17. Pouzaud, F., Bernard-Beaubois, K., Thevenin, M., Warnet, J. M., Hayem, G. and Rat, P., 2004. In vitro discrimination of fluoroquinolones toxicity on tendon cells; Involvement of oxidative stress. Journal of Pharmacology and Experimental Therapeutics, 308, 394-402.PubMedCrossRefGoogle Scholar
  18. Riley, G. P., Curry, V., DeGroot, J., van El, B., Verzijl, N., Hazleman, B. L. and Bank, R. A., 2002. Matrix metalloproteinase activities and their relationship with collagen remodeling in tendon pathology. Matrix Biology, 21, 185-195.PubMedCrossRefGoogle Scholar
  19. Sendzik, J., Shakibaei, M., Schafer-Korting, M. and Stahlmann, R., 2005. Fluoroquinolones cause changes in extracellular matrix, signaling proteins, metalloproteinase and caspase-3 in cultured human tendons cells. Toxicology, 212, 24-36.PubMedCrossRefGoogle Scholar
  20. Shakibaei, M. and Stahlmann, R., 2001. Ultrastructure of Achilles tendon from rats after treatment with fleroxacin. Archives of Toxicology, 75, 97-102.PubMedCrossRefGoogle Scholar
  21. Stahlmann, R. and Lode, H., 1999. Toxicity of quinolones. Drugs, 58, 37-42.PubMedCrossRefGoogle Scholar
  22. Stahlmann, R., Kuhner, S., Shakibaei, M., Schwabe, R., Flores, J., Evander, S. A. and van Sickle, D. C., 2000. Chondrotoxicity of ciprofloxacin in mature beagle dogs: immunohistochemistry, electron microscopy and drug plasma concentrations. Archives of Toxicology, 73, 564-572.PubMedCrossRefGoogle Scholar
  23. Stewart, M. C., Saunders, K. M., Burton-Wurster, N. and Macleod, J. N., 2000. Phenotypic stability of articular chondrocytes in vitro: the effects of culture models, bone morphogenetic protein 2, and serum supplementation. Journal of Bone Mineral Research, 15, 166-174.CrossRefGoogle Scholar
  24. Van der Linden, P. D., van de Lei, J., Nab, H. W., Knol, A. and Stricker, B. H., 1999. Achilles tendinitis associated with fluoroquinolones. British Journal of Clinical Pharmacology, 48, 433-437.PubMedCrossRefGoogle Scholar
  25. Van der Linden, P. D., van Puijenbroek, E. P., Feenstra J., Veld, B. A., 2001. Sturkenboom, M. C., Herings, R. M., Leufkens, H. G. and Stricker, B. H., 2001. Tendon disorders attributed to fluoroquinolones: a study on 42 spontaneous reports in the period 1988 to 1998. Arthritis and Rheumatism, 45, 235-239.PubMedCrossRefGoogle Scholar
  26. Van der Linden, P. D., Sturkenboom, M. C., Herings, R. M., Leufkens, H. G. and Stricker, B. H., 2002. Fluoroquinolones and risk of Achilles tendon disorders: case-control study. BMJ (Clinical Research education, London), 324, 1306-1307.CrossRefGoogle Scholar
  27. Van der Linden, P. D., Sturkenboom, M. C., Herings, R. M., Leufkens, H. M., Rowlands, S., Stricker, B. H., 2003. Increased risk of Achilles tendon rupture with quinolone antimicrobial use, especially in elderly patients taking oral corticosteroids. Archives Internal Medicine, 163, 1802-1807.Google Scholar
  28. Williams, R. J., Attia, E., Wickiewicz, T. L. and Hannafin, J. A., 2000. The effect of ciprofloxacin on tendon, paratenon, and capsular fibroblast metabolism. American Journal of Sports Medicine, 28, 364-369.PubMedGoogle Scholar
  29. Yoon, J. H., Brooks, R. L. Jr., Zhao, J. Z., Isaacs, D. and Halper, J., 2004a. The effects of enrofloxacin on decorin and glycosaminoglycans in avian tendon cell cultures. Archives Toxicology, 78, 599-608.CrossRefGoogle Scholar
  30. Yoon, J. H., Brooks, R. L. Jr., Khan, A., Pan, H., Bryan, J., Zhang, J., Budsberg, S. C., Mueller, P. O. and Halper, J., 2004b. The effect of enrofloxacin on cell proliferation and proteoglycans in horse tendon cells. Cell Biology Toxicology, 20, 41-54.CrossRefGoogle Scholar
  31. Yu, C. and Giuffre, B., 2005. Achilles tendinopathy after treatment with fluoroquinolone. Australasian Radiology, 49, 407-410.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • S. Lim
    • 1
  • M. A. Hossain
    • 1
    • 2
  • J. Park
    • 1
  • S. H. Choi
    • 1
  • G. Kim
    • 1
    Email author
  1. 1.Laboratory of Veterinary Surgery, College of Veterinary MedicineChungbuk National UniversityCheongjuSouth Korea
  2. 2.Department of Pathology and ParasitologyChittagong Veterinary and Animal Sciences UniversityChittagongBangladesh

Personalised recommendations