## 3 Results
### Osteogenic Differentiation
\begin{table}
\begin{tabular}{c c c c c c} \hline \hline
**ARS** & \multicolumn{2}{c}{_p-Value_} & **ALP** & \multicolumn{2}{c}{_p-Value_} \\ \hline NM vs. DM & ns & 0.998 & NM vs. DM & ns & \(>\)0.99 \\ NM vs. RRMPU + DM & *** & \(<\)0.0001 & NM vs. RRMPU + DM & ** & 0.02 \\ NM vs. RRMPA + DM & ns & 0.9549 & NM vs. RRMPA + DM & ** & \(<\)0.001 \\ NM vs. FSP + DM & *** & 0.0003 & NM vs. FSP + DM & *** & \(<\)0.001 \\ NM vs. BIO + DM & *** & \(<\)0.0001 & NM vs. BIO + DM & *** & \(<\)0.001 \\ NM vs. MTA + DM & ns & 0.9843 & NM vs. MTA + DM & *** & \(<\)0.001 \\ DM vs. RRMPU + DM & *** & \(<\)0.0001 & DM vs. RRMPU + DM & ** & 0.01 \\ DM vs. RRMPA + DM & ns & 0.9993 & DM vs. RRMPA + DM & *** & \(<\)0.001 \\ DM vs. FSP + DM & *** & 0.0008 & DM vs. FSP + DM & *** & \(<\)0.001 \\ DM vs. BIO + DM & *** & \(<\)0.0001 & DM vs. BIO + DM & *** & \(<\)0.001 \\ DM vs. MTA + DM & ns & \(>\)0.9999 & DM vs. MTA + DM & *** & \(<\)0.001 \\ RRMPU + DM vs. RBMPA + DM & *** & \(<\)0.0001 & RMMPU + DM vs. RBMPA + DM & *** & \(<\)0.001 \\ RRMPU + DM vs. FRMPA + DM & *** & \(<\)0.0001 & RRMPU + DM vs. RBMPA + DM & *** & \(<\)0.001 \\ RRMPU + DM vs. FSP + DM & *** & \(<\)0.0001 & RRMPU + DM vs. FSP + DM & *** & \(<\)0.001 \\ RRMPU + DM vs. FSP + DM & *** & \(<\)0.0001 & RRMPU + DM vs. FSP + DM & *** & \(<\)0.001 \\ RRMPU + DM vs. BID + DM & ns & 0.9959 & RRMPU + DM vs. BIO + DM & *** & \(<\)0.001 \\ RRMPU + DM vs. MTA + DM & *** & \(<\)0.0001 & RRMPU + DM vs. MTA + DM & *** & \(<\)0.001 \\ RRMPU + DM vs. MTA + DM & *** & \(<\)0.0001 & RRMPU + DM vs. MTA + DM & *** & \(<\)0.001 \\ RRMPA + DM vs. FSP + DM & *** & \(<\)0.0001 & RRMPU + DM vs. BID + DM & *** & \(<\)0.001 \\ RRMPU + DM vs. BID + DM & *** & \(<\)0.0001 & RRMPU + DM vs. BID + DM & *** & \(<\)0.001 \\ RRMPU + DM vs. BID + DM & *** & \(<\)0.0001 & RRMPU + DM vs. BID + DM & *** & \(<\)0.001 \\ RRMPU + DM vs. MTA + DM & *** & \(<\)0.0001 & BIO + DM vs. MTA + DM & ns & 0.1 \\ \hline \hline \end{tabular}
\end{table} 
Table 2: Comparative analysis of the tested bioceramic materials. ARS alizarin red staining; ALP alkaline phosphatase; NM normal medium, DM differentiation medium, RRMPU root repairing material purity, RRMPA root repairing material paste, FSP root repairing material purity fast set, BIO BiodentineTM, MTA mineral trioxide aggregate (ns = not significant, *\(p<0.033\), **\(p<0.002\), ***\(p<0.001\), ***\(p<0.0001\)). Figure 2: Alkaline phosphatase activity. Osteogenic differentiation was monitored by evaluation of alkaline phosphatase (ALP) activity: ALP levels were higher in MG-63 cells cultured in direct contact with FSP or RRMPU, compared with the cells grown in the presence of MTA. The ALP activity is expressed as absorbance at 405 nm. Figure 3: Formation of mineralized nodules. The presence of biomaterials enhanced the formation of mineralized nodules. (**a**) Mineralized nodules were detected in MG-63 cells grown in direct contact with RRMPU, BIO or FSP, as highlighted in red. (**b**) The area of nodules was increased in RRMPU, FSP and BIO cultures (\(p<0.001\)***). (**c**) Alizarin Red S staining demonstrated that biomineralization increased when cells were cultured in direct contact with RRMPU (\(p<0.0001\)***), BIO (\(p<0.0001\)***) or FSP (\(p=0.0012\)***). Alizarin Red S is expressed as absorbance at 405 nm (left) and as mM concentration (right).