title: Rotenone prevents impact-induced chondrocyte death.
abstract: Mechanical insult to articular cartilage kills chondrocytes, an event that may increase the risk of posttraumatic osteoarthritis. Recent reports indicate that antioxidants decrease impact-induced chondrocyte death, but the source(s) of oxidants, the time course of oxidant release, and the identity of the oxidative species generated in response to injury are unknown. A better understanding of these processes could lead to new treatments of acute joint injuries. To that end, we studied the kinetics and distribution of oxidant production in osteochondral explants subjected to a single, blunt-impact injury. We followed superoxide production by measuring the time-dependent accumulation of chondrocyte nuclei stained with the superoxide-sensitive probe dihydroethidium. The percentage of chondrocytes that were dihydroethidium-positive was 35% above baseline 10 min after impact, and 65% above baseline 60 min after impact. Most positive cells were found within and near areas contacted directly by the impact platen. Rotenone, an electron transport chain inhibitor, was used to test the hypothesis that mitochondria contribute to superoxide release. Rotenone treatment significantly reduced dihydroethidium staining, which remained steady at 15% above baseline for up to 60 min postimpact. Moreover, rotenone reduced chondrocyte death in impact sites by more than 40%, even when administered 2 h after injury (p < 0.001). These data show that much of the acute chondrocyte mortality caused by in vitro impact injuries results from superoxide release from mitochondria, and suggest that brief exposure to free radical scavengers could significantly improve chondrocyte viability following joint injury.
authors.affiliation: Department of Orthopaedics and Rehabilitation, 1182 ML, The University of Iowa, 500 Newton Rd., Iowa City, Iowa 52242, USA.
authors.forename: Wendy
authors.initials: W
authors.lastname: Goodwin
authors.forename: Daniel
authors.initials: D
authors.lastname: McCabe
authors.forename: Ellen
authors.initials: E
authors.lastname: Sauter
authors.forename: Eric
authors.initials: E
authors.lastname: Reese
authors.forename: Morgan
authors.initials: M
authors.lastname: Walter
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Buckwalter
authors.forename: James A
authors.initials: JA
authors.lastname: Martin
mesh_terms: D000818:Animals; D002358:Cartilage, Articular / Q000293:injuries; D002417:Cattle; D002470:Cell Survival / Q000187:drug effects; D019902:Chondrocytes / Q000187:drug effects* / Q000502:physiology; D016166:Free Radical Scavengers / Q000494:pharmacology; D008928:Mitochondria; D010003:Osteoarthritis / Q000378:metabolism; D012402:Rotenone / Q000494:pharmacology*; D013481:Superoxides / Q000378:metabolism*; D013977:Tibia / Q000293:injuries / Q000473:pathology*; D046509:Tissue Culture Techniques
chemical_list: D016166:Free Radical Scavengers; D012402:Rotenone; D013481:Superoxides