Effects of Adipokines and Insulin on Intracellular pH, Calcium Concentration, and Responses to Hypo-Osmolarity in Human Articular Chondrocytes from Healthy and Osteoarthritic Cartilage

OBJECTIVE To evaluate the effects of adipokines and insulin on intracellular calcium concentration ([Ca(2+)]i) and pH (pHi) in human articular chondrocytes from healthy (CHC) and osteoarthritic cartilage (COC). DESIGN pHi and [Ca(2+)]i were measured using BCECF and Fura-2 fluorometric probes in CHC and COC under control conditions and following a hypotonic shock. The effects of interleukin-1β (IL1β), tumor necrosis factor-α (TNFα), insulin, leptin, resistin, and adiponectin were assessed. RESULTS pHi was lower in COC than in CHC. Only IL1β β decreased pHi in both cell types; all the agents enhanced pHi recovery following an ammonium prepulse in CHC, effect that was attenuated by Na(+)-H(+) exchanger inhibitors, but they had no effect in COC. Hypotonic shock (HTS) caused a pHi increase, which was significantly smaller in COC. All the hormones attenuated this response and the effect of IL1β was greater. The basal [Ca(2+)]i was similar in COC and CHC; IL1β, TNFα, and insulin increased the [Ca(2+)]i, but leptin, resistin, and adiponectin did not. These effects were greater in COC. This [Ca(2+)]i increase was dependent on extracellular Ca(2+) and attenuated by Na(+)-Ca(2+) exchanger inhibitors. HTS caused a [Ca(2+)]i increase, which was inhibited by transient receptor potential vanilloid blockers and attenuated by all the hormones tested with the exception of adiponectin. CONCLUSIONS These findings may help explain the association between obesity and osteoarthritis, in which these hormones are altered. The responses of CHC and COC are different, which suggests that a modification of pH and Ca(2+) homeostasis is part of the osteoarthritis pathophysiology.