Figure 2: Selenium reverses myofibroblast differentiation of prostatic fibroblasts. Therapeutic targeting of myofibroblast dysregulation in fibrotic disease may be accomplished by promoting myofibroblast dedifferentiation to the nonactivated fibroblast/progenitor phenotype. (a) Methodology outline of selenium (Se)-mediated reversal of fibroblast-to-myofibroblast differentiation in primary human prostatic fibroblasts. Myofibroblast differentiation was induced with 1 ng/mL TGFβ1 or bFGF as mock control. After 72 hours, fresh media was added containing bFGF or TGFβ1 as before but supplemented with selenium (Se) as sodium selenite or vehicle equivalent. Cells were incubated for a further 48 hours before harvesting. Reversal of myofibroblast differentiation of primary human prostatic fibroblasts treated as outlined in (a) was verified by (b) Western blotting for the myofibroblast markers, -SMA, and IGFBP3 and (c) morphological analysis using phase contrast microscopy. (b) Induction of myofibroblast differentiation by TGFβ1 in the absence of Se (0 nM) is indicated by increased production of myofibroblast markers. However, both -SMA and IGFBP3 levels are reduced in the presence of Se in a dose-dependent manner. (c) At the morphological level, Se restores the thin, elongated and light refractive phenotype to cells predifferentiated with TGFβ1 (far right), whereas cells treated with TGFβ1 alone (center panel) exhibit the typical enlarged and flattened myofibroblast phenotype with visible actin-like filaments. (b-c) Images are representative of three independent experiments using primary cells isolated from different donors.