Archives of Toxicology

, Volume 78, Issue 10, pp 599–608 | Cite as

The effects of enrofloxacin on decorin and glycosaminoglycans in avian tendon cell cultures

  • Jung Hae Yoon
  • Randolph L. BrooksJr.
  • Jian Zeng Zhao
  • David Isaacs
  • Jaroslava HalperEmail author
Organ Toxicity and Mechanisms


Tendonitis and tendon rupture have been reported to occur during or following therapy with fluoroquinolone antibiotics. Though the pathogenesis is unknown, several studies suggest that fluoroquinolone antibiotics alter proteoglycan content in soft tissues, including tendons, and thereby alter collagen fibrillogenesis. To better understand the mechanism of action of fluoroquinolones, we studied the effects of enrofloxacin, a widely used fluoroquinolone in veterinary medicine, on avian tendon cell cultures established from gastrocnemius tendons from 18-day-old chicken embryos. We found that cell proliferation was progressively inhibited with increasing concentrations of enrofloxacin. This was accompanied by changes in morphology, extracellular matrix content and collagen fibril formation as detected by electron microscopy. We also observed a 35% decrease in the content of total monosaccharides in enrofloxacin-treated cells. The ratio of individual monosaccharides was also altered in enrofloxacin-treated cells. Enrofloxacin also induced the synthesis of small amounts of keratan sulfate in tendon cells. Moreover we observed enrofloxacin-induced changes in glycosylation of decorin, the most abundant tendon proteoglycan, resulting in the emergence of multiple lower molecular bands that were identifiable as decorin after chondroitinase ABC and N-glycanase treatment of extracts from enrofloxacin-treated cells. Medium conditioned by enrofloxacin-treated cells contained less decorin than did medium conditioned by control cells. We hypothesize that enrofloxacin induces either changes in the number of N-linked oligosaccharides attached to the core protein of decorin or changes in decorin degradation process. In conclusion, our data suggest that enrofloxacin affects cell proliferation and extracellular matrix through changes in glycosylation.


Cell proliferation Decorin Enrofloxacin Monosaccharides Morphological changes Tendon cells 



This study was supported by a grant from The University of Georgia Research Foundation. We thank Dr. W.L. Steffens and Ms. Mary Ard for performing electron microscopy, and Dr. S.C. Budsberg for helpful discussions. Special thanks have to go to Ms. Trina Abbeny for help with GC-MS analysis and to Dr. L.S. Leshin for help with statistical analysis. The experiments comply with the current US laws.


  1. Bailey RR, Kirk JA, Peddie A (1983) Norfloxacin-induced rheumatic disease. N Z Med J 96:590PubMedGoogle Scholar
  2. Bernard-Beaubois K, Hecquet C, Hayem G, Rat P, Adolphe M (1998) In vitro study of cytotoxicity of quinolones on rabbit tenocytes. Cell Biol Toxicol 14:283–292CrossRefPubMedGoogle Scholar
  3. Blumenkrantz N, Asboe-Hansen G (1973) New method for quantitative determination of uronic acids. Anal Biochem 54:484–489PubMedGoogle Scholar
  4. Brenneisen P, Briviba K, Wlaschek M, Wenk J, Scharffetter-Kochanek K (1997) Hydrogen peroxide (H2O2) increases the steady-state mRNA levels of collagenase/MMP-1 in human dermal fibroblasts. Free Radic Biol Med 22:515–524CrossRefPubMedGoogle Scholar
  5. Burkhardt JE, Hill MA, Carlton WW, Kesterson JW (1990) Histologic and histochemical changes in articular cartilages of immature beagle dog dosed with difloxacin, a fluoroquinolone. Vet Pathol 27:162–170PubMedGoogle Scholar
  6. Campbell MA, Winter AD, Ilic MZ, Handley CJ (1996) Catabolism and loss of proteoglycans from cultures of bovine collateral ligament. Arch Biochem Biophys 328:64–72CrossRefPubMedGoogle Scholar
  7. Danielson KG, Baribault H, Holmes DF, Graham H, Kadler KE, Iozzo RV (1997) Targeted disruption of decorin leads to abnormal collagen fibril morphology and skin fragility. J Cell Biol 136:729–743CrossRefPubMedGoogle Scholar
  8. Egerbacher M, Edinger J, Tschulenk W (2001) Effects of enrofloxacin and ciprofloxacin hydrochloride on canine and equine chondrocytes in culture. Am J Vet Res 62:704–708PubMedGoogle Scholar
  9. Endtz HP, Ruijs GJ, van Klingeren B, Jansen WH, van der Reyden T, Mouton RP (1991) Quinolone resistance in Campylobacter isolated from man and poultry following the introduction of fluoroquinolones in veterinary medicine. J Antimicrob Chemother 27:199–208PubMedGoogle Scholar
  10. Garcia Ovando H, Gorla N, Luders C, Poloni G, Errecalde C, Prieto G, Puelles I (1999) Comparative pharmacokinetics of enrofloxacin and ciprofloxacin in chickens. J Vet Pharmacol Therap 22:209–212CrossRefGoogle Scholar
  11. Hahn-Danton EA, Aimes RT, Quigley JP (2000) The isolation, characterization, and molecular cloning of a 75-kDa gelatinase B-like enzyme, a member of the matrix metalloproteinase (MMP) family. An avian enzyme that is MMP-9 like in its cell expression pattern but diverges from mammalian gelatinase B in sequence and biochemical properties. J Biol Chem 275:40827–40838CrossRefPubMedGoogle Scholar
  12. Halper J, Hu W, Kisaalita WS, Griffin A, Rowland GN (2000) Immunohistochemical detection of fibrillar collagens in tissue sections and in cultured cells. J Histotechnol 23:333–400Google Scholar
  13. Hascall VC, Kimura JH (1982) Proteoglycans: isolation and characterization. Meth Enzymol 82:769–799PubMedGoogle Scholar
  14. Hayem G, Carbon C (1995) A reappraisal of quinolone tolerability. Drug Safety 13:338–342PubMedGoogle Scholar
  15. Hayem G, Petit PX, Levacher M, Gaudin C, Kahn MF, Pocidalo JJ (1994) Cytofluorometric analysis of chondrotoxicity of fluoroquinolones antimicrobial agents. Antimicrob Agents Chemother 38:243–247PubMedGoogle Scholar
  16. Holtom PD, Pavkovic SA, Bravos PD, Patzakis MJ, Shepherd LE, Frenkel B (2000) Inhibitory effects of the quinolone antibiotics trovafloxacin, ciprofloxacin, and levofloxacin on oesteoblastic cells in vitro. J Orthop Res 18:721–727PubMedGoogle Scholar
  17. Huec JC, Schaeverbeke T, Chauveaux D (1995) Epicondylitis after treatment with fluoroquinolone antibiotics. J Bone Joint Surg 77-B:293–295Google Scholar
  18. Intorre L, Mengozzi G, Bertini S, Bagliacca M, Luchetti E, Soldani G (1997) The plasma kinetics and tissue distribution of enrofloxacin and its metabolite ciprofloxacin in the Muscovy duck. Vet Res Commun 21:127–136CrossRefPubMedGoogle Scholar
  19. Kashida Y, Kato M (1997) Characterization of fluoroquinolone-induced Achilles tendon toxicity in rats: comparison of toxicities of 10 fluoroquinolones and effects of anti-inflammatory compounds. Antimicrob Agents Chemother 41:2389–2393PubMedGoogle Scholar
  20. Kato M, Takada S, Kashida Y, Nomura M (1995) Histological examination on Achilles tendon lesions induced by quinolone antibacterial agents in juvenile rats. Toxicol Pathol 23:385–392PubMedGoogle Scholar
  21. Lennon DP, Carrino DA, Baber MA, Caplan AI (1991) Generation of a monoclonal antibody against avian small dermatan sulfate proteoglycan: immunolocalization and tissue distribution of PG-II (decorin) in embryonic tissues. Matrix 11:412–427PubMedGoogle Scholar
  22. Li W, Vergnes JP, Cornuet PK, Hassell JR (1992) cDNA clone to chick corneal chondroitin/dermatan sulfate proteoglycan reveals identity to decorin. Arch Biochem Biophys 296:190–197PubMedGoogle Scholar
  23. Lo YY, Conquer JA, Grinstein S, Cruz TF (1998) Interleukin-1 beta induction of C-fos and collagenase expression in articular chondrocytes: involvement of reactive oxygen species. J Cell Biochem 69:19–29CrossRefPubMedGoogle Scholar
  24. McDermott PF, Bodeis SM, English LL, White DG, Walker RD, Zhao S, Simjee S, Wagner DD (2002) Ciprofloxacin resistance in Campylobacter jejuni evolves rapidly in chickens treated with fluoroquinolones. J Inf Dis 185:837–840CrossRefGoogle Scholar
  25. McNeil M, Darvill AG, Aman P, Franzé LE, Albersheim P (1982) Structural analysis of complex carbohydrates using high-performance liquid chromatography, gas chromatography, and mass spectrometry. Meth Enzymol 83:3–45PubMedGoogle Scholar
  26. Nielsen P, Gyrdhansen N (1997) Bioavailability of enrofloxacin after oral administration to fed and fasted pigs. Pharmacol Toxicol 80:246–250PubMedGoogle Scholar
  27. Pan H, Halper J (2003) Regulation of heat shock protein 47 and type I procollagen expression in avian tendon cells. Cell Tissue Res 311:373–382. DOI 10.1007/s00441-003-0699-zGoogle Scholar
  28. Riddell C (1997) Developmental, metabolic and other noninfectious disorders. In: Calnek BW (ed) Diseases of poultry, 10th edn. Iowa State University Press, Ames, Iowa, pp 913–950Google Scholar
  29. Scott PG, Nakano T, Dodd CM, Pringle GA, Kuc IM (1989) Proteoglycans of the articular disc of the bovine temporomandibular joint ii. Low molecular weight dermatan sulfate proteoglycan. Matrix 9:284–292PubMedGoogle Scholar
  30. Shakibaei M, Stahlmann R (2001) Ultrastructure of Achilles tendon from rats after treatment with fleroxacin. Arch Toxicol 75:97–102PubMedGoogle Scholar
  31. Shakibaei M, Stahlmann R (2003) Ultrastructural changes induced by the des-F(6)-quinolone garenoxacin (BMS-284756) and two fluoroquinolones in Achilles tendon from immature rats. Arch Toxicol 77:521–526CrossRefPubMedGoogle Scholar
  32. Shakibaei M, Pfister K, Schwabe R, Vormann J, Stahlmann R (2000) Ultrastructure of Achilles tendons of rats treated with ofloxacin and fed a normal or magnesium-deficient diet. Antimicrob Agents Chemotherap 44:261–266CrossRefGoogle Scholar
  33. Shen LL (1993) Quinolone-DNA interaction. In: Hooper DC, Wolfson JS (eds) Quinolone antimicrobial agents. American Society for Microbiology, Washington, DC, pp 77–95Google Scholar
  34. Simonin MA, Gegout-Pottie P, Minn A, Gillet P, Netter P, Terlain B (1999) Proteoglycan and collagen biochemical variations during fluoroquinolone-induced chondrotoxicity in mice. Antimicrob Agents Chemother 43:2915–2921PubMedGoogle Scholar
  35. Stahlmann R, Zippel U, Förster C, Schwabe R, Shakibae M, Merker HJ, Borner K (1998) Chondrotoxicity and toxicokinetics of sparfloxacin in juvenile rats. Antimicrob Agents Chemother 42:1470–1475Google Scholar
  36. Trichilis A, Saranteas T, Potamianou A, Mourouzis C, Tesseromatis C (2003) Quinolone levels in serum and maxillofacial tissues under iburpofen co-administration following surgical trauma. J Musculoskel Neuron Interact 3:179–175Google Scholar
  37. Wells-Knecht MC, Lyons TJ, McCance DR, Thorpe SR, Baynes JW (1997) Age-dependent increase in ortho-tyrosine and methionine sulfoxide in human skin collagen is not accelerated in diabetes. J Clin Invest 100:839–846PubMedGoogle Scholar
  38. Williams RJ III, Attia E, Wickiewicz TL, Hannafin JA (2000) The effect of ciprofloxacin on tendon, paratenon, and capsular fibroblast metabolism. Am J Sports Med 28:364–369PubMedGoogle Scholar
  39. Winter AD, Campbell MA, Robinson HC, Handley CJ (2000) Catabolism of newly synthesized decorin by explant cultures of bovine ligament. Matrix Biol 19:129–138CrossRefPubMedGoogle Scholar
  40. Yoon JH, Brooks R Jr, Halper J (2002) Immunoblotting assays for keratan sulfate. Anal Biochem 306:298–304CrossRefPubMedGoogle Scholar
  41. Zabraniecki L, Negrier I, Vergne P, Arnaud M, Bounet C, Bertin P, Treves R (1996) Fluoroquinolone induced tendinopathy: report of 6 cases. J Rheumatol 23:516–520PubMedGoogle Scholar
  42. Zhu WH, Guo X, Villaschi S, Nicosia RF (2000) Regulation of vascular growth and regression by matrix metalloproteinases in the rat aorta model of angiogenesis. Lab Invest 80:545–555PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Jung Hae Yoon
    • 1
    • 2
  • Randolph L. BrooksJr.
    • 1
    • 3
  • Jian Zeng Zhao
    • 1
  • David Isaacs
    • 1
    • 4
  • Jaroslava Halper
    • 1
    Email author
  1. 1.The Soft Tissue Center, Department of PathologyCollege of Veterinary Medicine, The University of GeorgiaAthensUSA
  2. 2.Department of BiochemistryBlacksburgUSA
  3. 3.Dept. of Medicine/RheumatologyUniversity of California at San DiegoLa JollaUSA
  4. 4.University of Health Sciences College of Osteopathic MedicineKansas CityUSA

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