A sulfate-arsenical-ferruginous water affects apoptosis, oxidative stress and the gene expression of inflammatory mediators and of a panel of MicroRNA in IL-1β stimulated human osteoarthritic chondrocytes.

The beneficial effects of balneotherapy in osteoarthritis (OA) using the sulfate-arsenicalferruginous Levico water (LW) have been recognized by several clinical studies. However, the specific biological properties of LW are partially understood. The present study aimed at evaluating the ability of LW in the regulation of apoptosis, oxidative stress and of the gene expression of a panel of articular cartilage markers and MicroRNA (miRNA) in human osteoarthritic chondrocytes exposed to interleukin (IL)-1β. Chondrocytes were obtained from the femoral heads of patients with OA who underwent surgery for total hip prostheses. The cells were incubated for 24 h and 48 h with different concentrations of LW, alone or in combination with IL-1β (10 ng/ml). Apoptosis and mitochondrial superoxide anion production were detected by cytometry. Gene levels of antioxidant enzymes, B-cell lymphoma (BCL)2, cytokines [IL-1β, IL-6, tumor necrosis factor (TNF)-α], metalloproteinases (MMPs)1 and 13, type II collagen (Col2a1), aggrecan, and of a pattern of miRNA were evaluated by Quantitative Real-Time PCR analysis (PCR). The involvement of nuclear factor (NF)-κB was investigated by PCR and immunofluorescence and by pre-incubation with a specific inhibitor (BAY 117082, IKKα/β). LW at 50% or 25% concentration counteracted the negative effects of IL-1β on cell viability, apoptosis and mitochondrial superoxide anion production and on the gene expression of antioxidant enzymes, in a dose-dependent manner. The treatment of chondrocytes with 50% of LW significantly inhibited the gene expression of pro-inflammatory cytokines, MMPs and of miR-34a and miR-181a, while it upregulated miR-140 in IL-1β stimulated cells. The pre-incubation with BAY 11-7082, IKKα/β reduced the damage induced by IL-1β, and, interestingly, potentiated the properties of LW. Our data demonstrated the ability of LW to regulate apoptosis, oxidative stress and gene expression of main factors implicated in OA pathogenesis through a possible modulation of NF-κB signaling pathway. This study supports the use of balneotherapy with a sulfate-arsenical-ferruginous mineral water in the treatment of OA.