Effect of GILZ on TNF-α inhibition of MSC osteogenic differentiation.

<p>MSC-GFP and MSC-GILZ cells were cultured in osteogenic induction media (OS) in the absence or presence of indicated concentrations of TNF-α. (A) Cells were stained with alizarin red S on day 21 to visualize mineralized bone matrix. (B) Bar graph showing quantitative results of A. After scanning, the plates were destained with a solution containing 10% cetylpyridinium chloride in sodium phosphate. Concentrations of recovered ARS dye were determined by absorbance measurement and presented as fold changes relative to the OD value of MSC-GFP cells cultured in OS media. Each bar represents an average of 15 wells from 3 independent plates. (C) MSC-GFP and MSC-GILZ cells were cultured as above (A) for 8 days and stained for ALP to detect surface ALP-positive cell populations. (D) Bar graph showing quantitative results of C. Intensities of scanned images were analyzed with NIH Image J software and presented as fold changes relative to the value of MSC-GFP cells cultured in OS media. Each bar represents an average of 15 wells from 3 independent plates. Representative scanned images are shown. Error bars indicate S.D. a, <i>p</i><0.01.