## 3 Results
### Odonto/Osteogenic Differentiation of hDPSCs
#### 3.2.3 Elisa  
To evaluate the efficiency and maturation stage of odonto/osteoblast formation, ALP and OPN were measured on day 21 of differentiation (Figure 5). The data were consistent with the flow cytometry results; the BD-treated group showed a high secretion level of the measured markers ALP and OPN from day 1 to day 21 of treatment. Both ALP and OPN expression levels were elevated significantly (\(p<0.0005\) and \(p<0.003\), respectively) compared to day 1 of treatment.  
\begin{table}
\begin{tabular}{c c c c c c c} \hline \hline \multirow{2}{*}{**Symbol**} & \multicolumn{4}{c}{**Up\(-\)Down Regulation (Comparing to Control Group)**} \\ \cline{2-7}  & \multicolumn{2}{c}{**BD**} & \multicolumn{2}{c}{**Positive Control**} & \multicolumn{2}{c}{**Negative Control**} \\ \cline{2-7}  & \multicolumn{2}{c}{**Fold**} & \multicolumn{2}{c}{**Regulation**} & \multicolumn{1}{c}{**\(p\) Value**} & \multicolumn{1}{c}{**Fold**} & \multicolumn{1}{c}{**Regulation**} & \multicolumn{1}{c}{**\(p\) Value**} \\ \hline EGF & 3.90 & **0.005593** & 4.35 & **0.002983** & 2.45 & **0.025906** \\ \hline VDR & \(-2.85\) & **0.000005** & \(-1.02\) & 0.864395 & \(-1.19\) & 0.192859 \\ \hline CALCR & 6.55 & **0.029038** & 7.80 & **0.00001** & 5.32 & **0.001879** \\ \hline COMP & 7.15 & **0.026571** & **17.59** & 0.052347 & **23.30** & 0.075397 \\ \hline EGF & 3.90 & **0.005593** & 4.35 & **0.002983** & 2.45 & **0.025906** \\ \hline VDR & \(-2.85\) & **0.000005** & \(-1.02\) & 0.864395 & \(-1.19\) & 0.192859 \\ \hline CALCR & 6.55 & **0.029038** & 7.80 & **0.00001** & 5.32 & **0.001879** \\ \hline COMP & 7.15 & **0.026571** & **17.59** & 0.052347 & **23.30** & **0.075397** \\ \hline RUNX2 & \(-8.04\) & **0.000000** & \(-3.52\) & 0.062884 & \(-4.19\) & **0.000000** \\ \hline SMAD4 & \(-2.87\) & **0.001775** & \(-1.41\) & 0.141474 & \(-1.12\) & 0.789034 \\ \hline SMAD5 & \(-3.62\) & **0.000977** & \(-1.44\) & 0.124030 & \(-1.53\) & 0.086751 \\ \hline CSF2 & 5.12 & **0.046668** & 6.11 & **0.000051** & **4.16** & **0.009006** \\ \hline CSF3 & **4.17** & **0.041875** & 4.97 & **0.00003** & **3.39** & **0.004201** \\ \hline VEGFB & 1.95 & **0.000019** & 2.07 & **0.032152** & **3.61** & **0.000213** \\ \hline PDGFA & 2.32 & 0.062215 & 3.09 & 0.167867 & 4.63 & 0.098655 \\ \hline \hline \end{tabular}
\end{table}
Table 3: PCR array results of fold changes and the statistical significance of genes, which are involved in Ca\({}^{2+}\) signaling and regulatory genes. All groups were compared to day 1 of treatment. All groups were compared to day 1 of treatment. Red: upregulated genes, Green: Downregulated genes. The bold numbers represent significant \(p\) values.  
Figure 4: Flow cytometric results of the odonto/osteogenic differentiation markers. (**A**) Flow cytometric histograms, and (**B**) the statistical analysis of the Mean fluorescence Intensity (MFI) of flow cytometric results of the expression level of odonto/osteogenic markers; Osteocalcin (OCN) and Ostepontin (OPN), hDPSCs treated with: Biodentine, osteogenic differentiation media (positive control), compared to the uninduced cells cultured in their own growth media (negative control), after 3 weeks of treatment by flow cytometry. (***\(p<0.0005\), ***\(p<0.005\)).  
In comparison to the other control groups, the BD group showed the highest significant elevation in the secretion level of ALP, compared to the positive control (\(p<0.0005\)) and negative control (\(p<0.00005\)). For the OPN marker, the BD-treated group showed a significant increase in the secretion level of OPN, compared to the negative control (\(p<0.05\)), whereas the positive control showed the highest secretion level of the latter marker, compared to the BD-treated group (\(p<0.005\)) (Figure 5).