We report herein that cartilage proteolytic activity increased in bovine and rabbit articular cartilage after treatment with a purified staphylococcal culture medium or intraarticular infection with Staphylococcus aureus. Staphylococcal culture medium increased the release of gelatinolytic, collagenolytic, and caseinolytic activity into the medium of isolated chondrocytes or cartilage organ culture. The proteolytic activities were determined in assays using radiolabeled substrate and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Staphylococcal culture medium was proteolytically inactive by both assay techniques. RNA synthesis of isolated chondrocytes was unaffected by staphylococcal culture medium, whereas overall protein synthesis was inhibited by 84%. An analysis of extracts of Staphylococcus aureus-infected rabbit knee cartilage by substrate gels showed increased gelatinolytic and caseinolytic activity compared with extracts of uninfected knee cartilage. Our data suggest that the rapid loss of proteoglycan and persistent degradation of cartilage in staphylococcal septic arthritis is due to the production and activation of chondrocyte proteases.Septic arthritis continues to be a serious problem in urban medical centers (I ,2) and is recognized as a complicating factor in rheumatoid arthritis and other chronic arthritic conditions (3). Although most joint infections are acquired hematogenously, direct infection can result from deep wounds, surgical procedures, and intraarticular joint injections (2). In nongonococcal septic arthritis, Staphylococcus aureus is the organism most frequently found by culture (64%); in 25% of the patients positive for S aureus, the bacteria are methicillin resistant (1). Sterile "postinfectious" arthritis provides the potential for residual joint damage (43).In an animal model of staphylococcal infectious arthritis, cartilage degradation was shown to continue, despite early antibiotic treatment (5). Infection of rabbit knees with S aureus was shown to cause a rapid loss of cartilage proteoglycan (PG) (a), with a loss of as much as 40% of the total PG from the cartilage matrix within the first 48 hours after infection (8). Loss of collagen becomes significant between the second and third weeks after infection (8). Antibiotic treatment begun within 24 hours of infection decreases collagen loss but fails to prevent loss of PG from the matrix (5).In vitro, S aureus or S aureus-conditioned culture medium induces cartilage degradation in viable cartilage explants (9). Cartilage degradation is marked by a loss of partially degraded PG, which is consistent with increased proteolytic activity. PG release from cartilage by S aureus or staphylococcal culture medium is significantly reduced by freeze-thawing cartilage (9). Killed bacteria have no effect on cartilage degradation. Staphylococcus-induced PG release is dependent on chondrocyte metabolism and can be blocked by inhibitors of RNA and protein synthesis, i.e., actinomycin D and cycloheximide, respectively