Two fluoroquinolones and their combinations with hyaluronan: comparison of effects on canine chondrocyte culture

Abstract

Fluoroquinolones (FQs) are frequently used for septic arthritis. Increased antibacterial activity has been associated with mammalian cell cytotoxicity that may increase the risk of developing osteoarthritis. This study compared the direct effects of two different FQs, enrofloxacin (Enro) and marbofloxacin (Mar), on normal primary canine chondrocytes and inflammatory-stimulated chondrocytes, in addition to their administration in combination with hyaluronan (HA). Cell viability, cell apoptosis, s-GAG production, and expression patterns of inflammatory, extracellular matrix (ECM) component and protease genes were measured. Enro co-culturing with HA could modify s-GAG synthesis compared with the negative control group. Co-treatment with both FQs and HA significantly decreased cell viability and induced more total apoptotic cell death compared with the negative control and pre-IL-1β-stimulated group. Enro regulated IL-1β-stimulated cells to overexpress IL-1β, TNF, and MMP3, whereas Mar induced upregulation of PTGS2 and NFKB1 and enhanced the expression of ECM component genes HAS1, COL2A1, and ACAN as well as TIMP1 and MMP9. Simultaneous use of HA with Enro can effectively reduce the expression of IL-1β, TNF, and MMP3 in pre-IL-1β-stimulated chondrocytes. These results suggest the beneficial effects of HA in reducing the adverse effects of Enro treatment at the transcriptional level.

Comparison of the effects of cefazolin and ceftriaxone on canine chondrocyte culture

“…IL‐1β was used to study the effects of CEFs on preconditioned chondrocytes, mimicking the initial inflammatory environment from pro‐inflammatory bacterial compounds in joint infections (Roland & Helmut, ). Our previous study described the characteristics of primary canine chondrocytes after stimulation with IL‐1β, which induced changes in the expression of catabolic mediators, i.e., inflammation, cytokines/chemokines, and matrix degradation (Siengdee et al., ). Overall, we investigated the effects of two CEFs, with different molecular structures, on Non‐and IL‐1β‐stimulated normal and OA canine chondrocytes, which resulted in differences in toxicity induction, s‐GAG synthesis and gene expression level.…”

“…Chondrocyte apoptosis and necrosis were assessed using a Muse® Caspase‐3/7 Assay Kit and Muse® Cell Analyzer (Merck Millipore, Darmstadt, Germany) following the manufacturer's instructions for flow cytometry, as previously described (Siengdee, Euppayo, Buddhachat, Chomdej, & Nganvongpanit, ). Briefly, the cells were trypsinized to produce a single‐cell suspension using 0.25% trypsin/EDTA.…”

Post-treatment of hyaluronan to decrease the apoptotic effects of carprofen in canine articular chondrocyte culture

“…Our previous studies involving canine chondrocyte cultures in vitro produced a different result when HA was combined with other drugs. However, HA could lessen the side effects of fluoroquinolones when treatment was given in conjunction with those other drugs (Siengdee et al, 2016). On the contrary, the therapeutic effects of HA could not mitigate the side effects of carprofen (Euppayo et al, 2015), corticosteroid (Siengdee et al, 2015) and triamcinolone acetonide (Euppayo et al, 2016) when subjects were treated with HA at the same time.…”

“…Several drugs, such as local anesthetic drugs (10 mg/mL of lidocaine or 7.5 mg/mL of ropivacaine in human chondrocyte culture), corticosteroids (25 mm of dextramethasone, 25 mg/mL of prednisolone or 7 mg/mL of betamethasone in human chondrocyte culture), cefazolin (2.0-3.33 mg/mL in canine chondrocyte culture) and fluoroquinolones (182 mg/mL of enrofloxacin or 295 mg/mL of marbofloxacin), have been shown to produce adverse effects on articular cartilage including apoptosis and the inhibition of growth of chondrocyte cells (Chrysis et al, 2005;Farkas et al, 2010;Nakazawa et al, 2002;Siengdee et al, 2016;Siengdee et al, 2017). This phenomenon can result in a loss of extra-cellular matrix formation and the progressive degradation of articular cartilage (Farkas et al, 2010).…”

“…For example, antibiotics such as cefazolin (2.0-3.33 mg/mL) may cause apoptosis and necrosis of chondrocytes by inducing the alpha tumor necrosis factor (TNF-a) and matrix metelloproteinase (MMP) production, decreasing the expression of key matrix molecules of the cartilage encoded by aggrecan (ACAN) and type II collagen COL2A1, and by downregulating the tissue inhibitors of the matrix metalloproteinase-1 (TIMP1) (Siengdee et al, 2017). Fluoroquinolone treatment has also been widely associated with certain side effects, including the development of severe musculoskeletal disorders, for example, tendinitis and tendon ruptures in humans after a 1-week course of ciprofloxacin, 250 mg twice daily (Akali & Niranjan, 2008), cartilage deformities after receiving ofloxacin, 20 mg/kg once daily for 7 consecutive days in 3-month-old male beagle dogs (Yabe et al, 1997), along with the altering of inflammatory mediators and extracellular matrix (ECM) components, as well as the MMP encoding genes in chondrocyte cultures in vitro (Siengdee et al, 2016). Non-steroidal anti-inflammatory drugs (NSAIDs), including indomethacin, ketorolac, diclofenac, piroxicam and celecoxib, caused proliferation suppression and the cell death of chondrocyte culture in vitro (Chang et al, 2006).…”