Purpose: Increase of proinflammatory cytokines in the synovial fluid is associated with articular cartilage degradation and believed to be one of the contributing factors of osteoarthritis (OA) progression. Previous studies reported that Lactobacillus casei Shitora consumption was associated with reduced pain and decreased joint damage in OA knee patients but barely suggested mechanisms underlying this therapeutic effect. We aim to investigate the anti-inflammatory effect of L. casei L39 (LC-L39) in a model of inflammation induced by lipopolysaccharide (LPS) in human chondrocytes through cytokine analysis. Methods: Human osteoarthritic chondrocytes were harvested from the articular cartilage of knee OA patients undergoing total knee replacement. Lactobacillus conditioned media (LCM) were prepared as previously described. Cells were cultured on 24-well plates to 70-80% confluence, then stressed by the addition of 20 ng/ml LPS with 0%, 5% and 10% LCM co-treatments. After 6 hours, the levels of secreted cytokines (TNF-a, IL-6 and IL-8) in the supernatants were determined using ELISA. Cell viability was evaluated by Trypan blue dye exclusion assay. Cell viability above 80% was considered non-cytotoxic. The Results were displayed as mean values ± S.D. Statistical analyses were performed using one-way analysis of variance (ANOVA) and Student's t-test to compare cytokine levels among groups. Results: Baseline TNF-a, IL-6 and IL-8 levels detected in unstimulated chondrocytes from OA donors were minimally detectable. In response to 20 ng/ml LPS stimulation, TNF-a, IL-6 and IL-8 levels were significantly increased. Treatment with 5% and 10% LCM resulted in the significantly dose-dependent reduction (p < 0.05) of IL-6 and IL-8 levels. With 10% LCM treatment, IL-6 was suppressed to baseline level. Interestingly, treatment with 5% LCM had no effect in reducing the level of TNF-a level but treatment with 10% LCM significantly reduced the level of TNF-a (p < 0.05). Cell viability was > 80% in all experiments. Conclusions: Our data demonstrates the anti-inflammatory effect of LC-L39 in a model of LPS-induced inflammatory cytokine production in human chondrocytes. This finding supports the potential of LC-L39 to alleviate articular cartilage degradation in knee OA. Further study is needed to understand the role of LC-L39 in cessation of OA perpetuation and give insight into novel disease-modifying therapeutic strategy.