Effect of levofloxacin on glycosaminoglycan and DNA synthesis of cultured rabbit chondrocytes at concentrations inducing cartilage lesions in vivo

Abstract

We investigated the toxic effect of levofloxacin (LVFX), a quinolone antibacterial agent, on cartilage by examining aspects of its in vivo toxicokinetics and effect on the function of cultured chondrocytes of the femoral articular cartilage from juvenile New Zealand White rabbits. Repeated administration of LVFX (100 mg/kg) orally for 7 days induced focal necrosis and superficial erosion in the articular cartilage of the femoral condyle, but 30 mg/kg did not. Concentrations of LVFX in the cartilage were highest at the first sampling point (30 min) after a single administration, being 4.93 and 12.2 g/g in the 30-and 100-mg/kg groups, respectively. The arthropathic concentration of LVFX in the cartilage was then shown to be 12.2 g/g or more. For an in vitro study, chondrocytes were separated from the articular cartilage of the rabbit femoral condyle and cultured for 7 days until confluence. 35SO 4 uptake by cultured chondrocyte sheets was most susceptible to LVFX, decreasing at drug concentrations of 5 g/ml or more in 24-and 48-h cultures but not in a 72-h culture. Furthermore, 3 H-thymidine uptake was decreased at concentrations of 10 g/ml or more in a 48-h culture but not in 24-and 72-h cultures. Rhodamine 123 accumulation was susceptible to inhibition in cultured chondrocytes at an LVFX concentration of 10 g/ml or more. These results suggest that LVFX inhibits glycosaminoglycan synthesis initially and DNA synthesis and mitochondrial function secondarily at actual arthropathic concentrations in cultured rabbit chondrocytes but that these changes are reversible and not enough to kill the cells.Adverse effects of quinolone antibacterial drugs, including arthralgia, joint swelling, arthropathy, and arthritis, have been reported at incidences of less than 0.5% in healthy volunteers and juvenile and adult patients (17). Quinolones are well known to induce cavitation in the articular cartilage of juvenile rats, rabbits, guinea pigs, dogs, marmosets, and crab-eating monkeys but not in that of adults; of these species, dogs are thought to be most susceptible to this chondrotoxicity (4,5,11,12,15,16,24,25,27,29,31). However, the relationship between the adverse clinical effects of these drugs and their toxic lesions in laboratory animals has not been clarified.The initial changes in quinolone-induced toxicity in the articular cartilage of juvenile animals are thought to occur in chondrocytes (6,8,19) or in the matrix (4). In our previous ex vivo study, ofloxacin (OFLX) inhibited the uptake of 3 H-thymidine and 35 SO 4 by the articular cartilage of juvenile rats 12 h after oral administration of a single large dose (20). Considering the inhibitory effect of the drug on topoisomerase II of mammalian cells, we then speculated that the initial target of OFLX in its induction of cartilage damage was the DNA synthesis of chondrocytes. Histopathological examinations by Stahlmann et al. (29) and Burkhardt et al. (6,8) after administration of OFLX and difloxacin revealed mitochondrial swelling and distention of the rough endopla...

Genistein-mediated inhibition of glycosaminoglycan synthesis as a basis for gene expression-targeted isoflavone therapy for mucopolysaccharidoses

“…18,19 Thus, we assumed that genistein may inhibit synthesis of GAGs owing to impairment of expression of genes coding for one or more of enzymes involved in this process. Some data obtained by others in experiments on cancer cells could suggest that this might be the case; 20 -22 23 We found that genistein inhibited GAG synthesis significantly in both wild-type and MPS cells (Figure 1). The most effective inhibition was observed at concentrations of genistein about 10 -30 mM, and further increase in concentration of this isoflavone did not cause more pronounced effects (Figure 1 and data not shown).…”

Chondrotoxicity of Quinolone Antimicrobial Agents

“…In an in vitro study using juvenile dog chondrocytes, nalidixic acid, ciprofloxacin, and three other quinolones inhibited mitochondrial dehydrogenase activity with inhibited proteoglycan synthesis 50 . Compromised mitochondrial integrities have also been shown in other in vitro experiments 51,52 . Changes in glycosaminoglycan and DNA synthesis and fibronectin-A single administration of pefloxacin to mice decreases biosynthesis of proteoglycan for the first 24 h, with recovery 48 h later.…”

“…The decrease is also observed ex vivo, suggesting a direct effect of pefloxacin on this process, and a 10-day treatment induces oxidative damage in collagen, which may be induced by oxygen-derived reactive species in the extracellular matrix 53 . Moreover, in many in vitro and ex vivo studies, the cartilage of various animal species has exhibited inhibition of synthesis of either collagen or glycosaminoglycans 28,29,50,51,[54][55][56][57] or proteoglycan and DNA 28,[50][51][52]56 . An increase in fibronectin staining is observed in the vicinity of the cavity in the articular cartilage of juvenile dogs and rats receiving difloxacin and ofloxacin, respectively 41,42 .…”

A Non-Arthropathic Dose and Its Disposition Following Repeated Oral Administration of Ofloxacin, a New Quinolone Antimicrobial Agent, to Juvenile Dogs.

“…Supporting this hypothesis, several DNA synthesis inhibitors have been stated to increase in the incidence of cartilage lesions induced by the quinolone in rats [18]. In a more recent investigation using the cultured rabbit chondrocyte at concentrations inducing cartilage lesions, the quinolone inhibited glycosaminoglycan synthesis initially and DNA synthesis and mitochondrial function secondarily [11]. However, the precise mechanism of quinoloneinduced arthropathy remains unclear.…”

“…However, quinolones have been reported to induce arthropathy in juvenile animals such as mice [12], rats [4,9,10], rabbits [11], dogs [3,5,8,20], nonhuman primates [17] and others [1,2] as a class effect of these derivatives. Among these species, the juvenile dog is thought to be most susceptible to articular cartilage lesions [6,19,20].…”