FIG. 1.

Long-term effect of diclofenac sodium on response of primary calvarial osteoblasts in proinflammatory cytokine medium. Osteoblasts were cultured for 14 days in osteogenic medium before experimentation. Cytokines (0.25 ng/mL interleukin-1β [IL-1β], 2.5 ng/mL tumor necrosis factor-α [TNF-α], and 25 ng/mL interferon-γ [IFN-γ]) were applied throughout the 27-day period. (A) Cell viability, converted to percentage of control medium reading. Values represented as mean±SD, n=6. Experiment repeated in triplicate. (B) Cumulative nitrite concentration in culture medium at timepoints. Values represented as cumulative mean±cumulative SD, n=6. Experiment repeated in triplicate. (C) Prostaglandin E₂ (PGE₂) concentration in medium at timepoints. Values represented as cumulative mean±cumulative SD. Experiment repeated in triplicate. For (A–C): statistical significance versus Control, *p≤0.01. Statistical significance versus IL-1β +TNF-α +IFN-γ, ^#p≤0.01. (D) Representative immunocytochemistry images showing expression of osteopontin (OPN)/osteocalcin (OCN) and cadherin-11 (cad-11)/collagen-I (col-I) after treatment with IL-1β, TNF-α, and IFN-γ and diclofenac sodium. Scale bar=90 μm. OCN (red), OPN (green), OCN/OPN (Hoescht 33258 nuclear stain [blue]), Cad-11 (red), and Col-I (green), Cad-11/Col-I (Hoescht 33258 nuclear stain [blue]). Color images available online at www.liebertpub.com/tea