Figure 3.

Cadmium inhibits the osteogenic differentiation of BMMSCs. BMMSCs were exposed to 0, 0.1, or 0.2 μM CdCl₂ for 1, 3, or 7 days. (A–E) The mRNA levels of the osteogenic differentiation markers, such as alkaline phosphatase (ALP) (A), osteocalcin (OCN) (B), runt-related transcription factor 2 (Runx2), (C), osterix (OSX) (D), and osteopontin (OPN) (E) were measured by quantitative Reverse-Transcriptase Polymerase Chain Reaction (qRT-PCR) (mean ± SD, n = 3). * p < 0.05, ** p < 0.01, different from control BMMSCs. BMMSCs were exposed to 0, 0.1, or 0.2 μM CdCl₂ for 7 days. (F) and (G) The mRNA levels of the osteogenic differentiation markers, ALP, OCN, Runx2, OSX, and OPN were determined by RT-PCR analysis (mean ± SD, n = 3). * p < 0.05, ** p < 0.01, different from control BMMSCs. BMMSCs were exposed to 0, 0.1, or 0.2 μM CdCl₂ for 1, 3, or 7 days. (H) ALP activity was detected by ALP assay (mean ± SD, n = 3). * p < 0.05, ** p < 0.01, different from control BMMSCs. BMMSCs were exposed to 0, 0.1, or 0.2 μM CdCl₂ for 7 days. (I) Western blots were performed, and (J) relative protein levels of ALP and Runx2 were determined (mean ± SD, n = 3). * p < 0.05, ** p < 0.01, different from control BMMSCs. BMMSCs were exposed to 0, 0.1, or 0.2 μM CdCl₂ and subjected to osteogenic differentiation for 14 days. (K) The ALP content and the numbers of mineralization nodules were evaluated by ALP staining (upper) and alizarin red S staining (lower). Scale bar, 100 μm.