Objective: To evaluate the anti-inflammatory effects of clinically relevant naproxen sodium (Nx) concentrations on human monocyte-derived macrophages in a controlled in vitro system and human primary synovial fluid (SF) cells. Design: Using phorbol 12-myristate 13-acetate, THP-1 human monocytic cells were differentiated into mature monocyte-derived macrophages in vitro then treated with Nx pre-or post-activating an inflammatory response with lipopolysaccharide (LPS) and hyaluronan (HA) fragments (n ¼ 8/group). Cell culture supernatants were assessed for NF-kB activity and prostaglandin E 2 (PGE 2 ), indicating cyclooxygenase enzyme activity. Under Duke IRB approval, primary human SF cells were collected at the time of knee joint replacement (n ¼ 19 individuals) for osteoarthritis (OA), and cultured with LPS, HA and Nx; SF cells were characterized by polychromatic flow cytometry for cell surface markers and intracellular cytokines. Result: Compared to placebo treatment of THP-1 cells, low dose Nx (corresponding 27.5e440 mg/L orally) added both pre-and post-activation with LPS/HA, significantly reduced NF-kB activity and PGE2: mean reduction to 73%, 61%, 17% and 10% of placebo, respectively. LPS/HA treatment of primary OA SF cells significantly increased the number of IL-1b producing primary monocytes and macrophages, and by 24 h the overall production of secreted cytokines (IL-1b, IL-6, IL8, and TNF-a). Low dose Nx reduced the percentage of IL-1b producing primary monocytes and macrophages. Conclusion: LPS/HA induced inflammation of THP-1 monocytic and primary human SF cells. Low dose Nx both prevented and reduced inflammatory responses of a human monocytic cell line and reduced IL-1b production by primary human SF monocytes and macrophages.