In knee osteoarthritis (OA), concentrations of interleukin (IL)-1β and tumor necrosis factor (TNF)-α increase in joint tissues and synovial fluid which incite a catabolic cascade and further the progression of OA. Several microRNAs (miRNA) have been associated with apoptosis (miR-16), inflammation (miR-22, miR-146a), and matrix degradation (miR-140, miR-27b) in developed OA or its symptoms. In this study, the time- and concentration-dependent nature of cellular and extracellular miRNAs in synoviocytes, meniscus cells, and chondrocytes as influenced by inflammatory cytokines was investigated. For time-dependent studies, three cell types were stimulated with 10 ng/ml IL-1β or 50 ng/ml TNF-α for 8, 16, and 24 h. For concentration-dependent studies, chondrocytes were stimulated with a higher level of IL-1β (20 ng/ml) or TNF-α (100 ng/ml) for 8 h. Cellular and extracellular expressions of miR-22, miR-16, miR-146a, miR-27b, and miR-140 were analyzed by RT-PCR. Time-dependent cellular miRNA expressions were similar across the three cell types with miR-146a significantly up-regulated and miR-27b significantly down-regulated at all time points. However, chondrocytes exhibited a unique extracellular miRNA profile with an increased release rate of miR-27b at 24 h. Our findings support further research into the characterization of miRNAs in synovial fluid for the development of early detection strategies of OA or cartilage injury. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:779–790, 2016.