Using an in vitro model, based on primary cultured chondrocytes, we examined possible oxidative injury caused by activated polymorphonuclear neutrophil granulocytes (PMNs), which are thought to be part of the pathomechanism of hemarthrosis. Chondrocytes were isolated from swine knee joints and divided into three groups. Pure chondrocytes acted as the control population (group I). PMNs from the systemic circulation, and hydrogen peroxide (as an artificial source of reactive oxygen species (ROS)) were added to groups II and III, respectively. All cultures were incubated for 6 hours. After the experiment, lipid membrane degradation by ROS was assessed by monitoring changes in the levels of malondialdehyde (MDA) and 4-hydroxyalkenal contents of the chondrocyte specimens. Changes in the endogenous scavenger status of the chondrocytes were characterized by measuring of reductions in glutathione (GSH) concentration and superoxide dismutase (SOD) activity. Significant increases in MDA/4-hydroxyalkenal levels and SOD activity as well as an expressive reduction in intracellular GSH content were highlighted by comparing the control to the PMN- or H2O2-treated cell populations. These findings confirm previous suggestions that PMN-derived ROS contribute to degradation of cartilage in hemarthrosis.
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