Osteogenesis from Dental Pulp Derived Stem Cells: A Novel Conditioned Medium Including Melatonin within a Mixture of Hyaluronic, Butyric, and Retinoic Acids

Figure 1

Effect of melatonin and HA + BU + RA exposure on the expression of genes orchestrating hDPSCs commitment toward an osteogenic lineage. Cells were exposed for 1, 3, 7, 14, or 21 days in the presence of one of the 4 conditioned media described under Materials and Methods and based on 100 nM dexamethasone with 200 μM L-Ascorbic acid 2-phosphate (medium D), 0,01 M melatonin (medium M), or a mixture of HA 2 mg/mL + BU 5 mM + RA 1 μM (medium H) or on a combination of melatonin together with HA + BU + RA (medium MH) or kept at the undifferentiated state by growing on medium C. The amounts of vascular endothelial growth factor A (VEGF A), Runx2, ZBTB16, and NR4A3 mRNA from treated or untreated cells were normalized to GAPDH, and the mRNA expression of cells differentiated with the 4 different conditioned media (D, M, H, and MH) was plotted at each time point as fold change relative to the expression in control undifferentiated cells defined as 1 (mean ± SE; ). All data from medium D-treated cells, M-treated cells, medium H-treated cells, and medium MH-treated cells at each time point were significantly different from those in control undifferentiated cells. mRNA levels from cells that had been exposed to medium D, medium M, medium H, and medium MH were significantly different from each other at each time point. Significantly different from medium D-treated cells. Medium D: green bars, medium M: purple bars, medium H: blue bars, and medium MH: orange bars.