Fig 9.

TNF-α is involved in inhibitory effects induced by supernatants from B. abortus-infected macrophages. Pretreatment of supernatants from B. abortus-infected macrophages harvested at 24 h p.i. with a neutralizing anti-TNF-α antibody inhibited the effects of the supernatants on osteoblast mineralization and organic matrix deposition. (A to D) Alizarin red S staining (A), ALP activity revealed with BCIP-NBT (B), Sirius red staining (C), and osteocalcin deposition revealed by immunofluorescence (D). (E to G) Quantitative analysis of alizarin red S (E), relative ALP activities (F), and Sirius red staining (G). *, P < 0.1 versus the results for unstimulated cells (Control). Data shown are from a representative experiment of five performed.