Figure 6
The effect of Mg100 conditioned media on the expression of osteoblast related genes and ALP levels. Fold change in the gene expression of (A) Runx-2 and (B) osteocalcin in cells cultured in filtered or non-filtered medium over a period of 12 days was investigated. On day 2, a significant fold change (*p < 0.05) in the expression of Runx-2 was observed when cells were cultured in Mg100 non-filtered medium compared to osteogenic medium. On day 4, a significant fold change (**p < 0.01) in osteocalcin expression was also observed when cells were cultured in Mg100 non-filtered medium compared to Mg100 filtered medium and osteogenic medium. On day 8 and 12, a similar trend was also observed for osteocalcin expression (*p < 0.05). (C) The expression of COL1A1 at day 2 was significantly downregulated when cells were cultured in non-filtered medium compared to filtered medium (*p < 0.05) and osteogenic medium (#p < 0.001). Fold change is relative to the control, represented by a fold change of 1. (D) The presence of ALP protein was evaluated at different time points (day 2, 7, 12) following culture in Mg 100 filtered and non-filtered medium. On day 12, a significant increase (#p < 0.001) in the presence of ALP was observed when cells were cultured in Mg100 non-filtered medium compared to Mg100 filtered medium and control. However, a significant reduction (#p < 0.001) in ALP presence was also observed when cells were cultured in Mg100 non-filtered medium compared to osteogenic medium. ALP protein was normalised to DNA concentration. Osteogenic medium was used as the positive control. The bars represent the mean and standard deviation in the positive orientation of three independent experiments, each with n = 3.
