Background Intra-articular injections of local anesthetics are used commonly in articular surgery. However, chondrocyte viability and metabolism may be adversely affected by various anesthetics. Objectives To assess the chondrotoxic effects of bupivacaine, levobupivacaine, and ropivacaine on human chondrocytes and elucidate possible mechanisms of chondrocyte death. Methods Cultured human chondrocytes (CHON-001) were exposed to 0.25% or 0.5% of bupivacaine, levobupivacaine, and ropivacaine in vitro. Cell viability was determined by flow cytometry after 15, 30, 60, and 120 min of exposure. Chondrocyte reactive oxygen species (ROS) production was measured every 10 min for up to 1 h using 2ʹ,7ʹ-dichlorodihydrofluorescein staining. Chondrocyte production of glycosaminoglycan was measured by capillary electrophoresis. NO production was measured using a colorimetric assay kit. Results We found a significant increase in chondrotoxicity dependent on exposure time and concentration of the anesthetic. At 60 min, chondrocyte viability was significantly (P < 0.05) decreased when exposed to 0.5% levobupivacaine (32.5%), or 0.25% or 0.5% bupivacaine (34.3% or 46.5%, respectively) compared with exposure to phosphate-buffered saline (PBS) vehicle as a control. Cell death at 120 min was mainly necrosis. There was no difference in viability after treatment with either concentration (0.25% or 0.5%) of ropivacaine at any time compared with exposure to PBS. We found increased production of NO, while ROS decreased after exposure to any of the anesthetics tested. Conclusions Ropivacaine may be safer than bupivacaine or levobupivacaine as an intra-articular analgesic. Chondrotoxicity of anesthetics in vitro may be mediated via a reactive nitrogen species-dependent pathway.

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