WSM up-regulates osteoblast differentiation markers <i>OCN</i> and <i>COL-1-A2</i> and accelerates bone nodule formation in MC3T3-A1 cells.

<p><b>A (a-b)</b>: Detection of transcriptional levels of osteoblast differentiation marker genes <i>viz</i>. osteocalcin (<i>OCN</i>) and pro-alpha 2(I) collagen (<i>COL-1A2</i>) in RT-PCR of MC3T3-A1 cells, with and without WSM (0.025% w/v) treatment for 24 h. The expression of <i>GAPDH</i>, as housekeeping gene was qualitatively (a) detected by the presence of bands in all experiments. Significant increase in transcript (b) of both the marker genes was observed after WSM treatment. <b>B (a-c)</b>: Alizarin red S staining of WSM treated (b) and control (a) MC3T3-E1 cells indicating WSM (0.025%, w/v) accelerates calcium deposition and enhances nodule formation. Black arrows indicate the mineral nodules. Quantification (c) of staining intensities were consistent with the visual observation. <b>C (a-c)</b>: von Kossa-von Giesen staining of WSM treated (b) and control (a) MC3T3-E1 cells indicating WSM (0.025% w/v) enhances nodule formation. Black arrows indicate the mineral nodules deposition. Quantification (c) of staining intensities were consistent with the visual observation. Results are expressed as mean -SEM (n= 3). *<i>p ≤ 0.05 (vs. control)</i>.