# The Impact of Silver Nanoparticles on Detntinal Tubule Penetration of Endodontic Bioceramic Sealer
## 3 Results
### The Evaluation of Sealer Penetration  
According to Kappa statistics, there was a very good agreement regarding the images analysis between the two evaluators (Kappa = 0.897). The dentinal tubule penetration values for the experimental groups are shown in Table 1 and presented in Figure 3. One-way ANOVA tests revealed a significant difference among the groups (\(p\) = 0.000) for both the dentinal tubule depths and total area of sealer penetration. For the dentinal tubule depth analysis, the Tukey multiple comparison test showed that Group 2 (BC+AgNPs/SC) had the significantly highest mean depth (2096 \(\pm\) 0.237 \(\upmu\)m), followed by Group 1 (BC/SC) (1506 \(\pm\) 0.362 \(\upmu\)m), and Group 4 (BC+AgNPs/CWC) (1417 \(\pm\) 0.503 \(\upmu\)m) with no significant difference. However, Group 3 (BC/CWC) had the significantly lowest mean depth (1108 \(\pm\) 0.260 \(\upmu\)m). For the total area of sealer penetration, the Tukey multiple comparison test showed that Group 2 (BC+AgNPs/SC) had the significantly highest mean value (5803 \(\pm\) 2.479 \(\upmu\)m\({}^{2}\)), followed by Group 4 (BC+AgNPs/CWC) (4285 \(\pm\) 3.153 \(\upmu\)m\({}^{2}\)). However, Groups 1 (BC/SC) (3107 \(\pm\) 3.112 \(\upmu\)m\({}^{2}\)) and 3 (BC/CWC) (2546 \(\pm\) 2.992 \(\upmu\)m\({}^{2}\)) had the lowest mean value, with no significant difference. Representative confocal laser scanning microscopic images are illustrated in Figure 4.  
Figure 3: The bar chart shows the mean \(\pm\) standard deviations of the maximum depth of penetration (**A**) and total area of sealer penetration (**B**) of the experimental groups.  
Figure 2: (**A**) SEM image of AgNPs illustrating the spherical shape and size range (\(\times\)200,000); (**B**) TEM image of AgNPs illustrates the spherical-shaped particles, as well as the absence of nanoparticles agglomeration (\(\times\)300,000); (**C**) TEM image of AgNPs mixed with biocearamic sealer demonstrates the spherical shape of AgNPs precipitated within the structure of the sealer, with no obvious agglomeration (\(\times\)150,000).  
Figure 4: Representative CLSM images showing the penetrability of the experimental sealers into denntinal tubules. (**A1**–**A4**) represent the BC+AgNPs/SC group, showing heavy and homogenous fluorescence, indicating great sealer penetration with long tags from the canal lumen toward the cementodentinal junction. (**B1**–**B4**) represent the BC+AgNPs/CWC group, showing sealer penetration through the denntinal tubules with long tags. (**C1**–**C4**) represent the BC/SC group, showing minimal sealer penetration into denntinal tubules with short tags. (**D1**–**D4**) represent the BC/CWC group, denoting hardly noticeable fluorescence, indicating minimal sealer penetration into denntinal tubules. All images are shown at 10\(\times\) magnification. All bars represent 200 \(\mu\)m.  
\begin{table}
\begin{tabular}{c c c c} \hline \hline
**Parameter** & **Group** & **Mean\(\pm\)SD** & _p-Value_ \\ \hline \multirow{3}{*}{Depth (\(\mu\)m)} & BC/SC & \(1506\pm 0.362\)\({}^{\text{b}}\) & \\ \cline{2-3}  & BC+AgNPs/SC & \(2096\pm 0.237\)\({}^{\text{c}}\) & \\ \cline{2-3}  & BC/CWC & \(1108\pm 0.260\)\({}^{\text{a}}\) & \\ \cline{2-3}  & BC+AgNPs/CWC & \(1417\pm 0.503\)\({}^{\text{b}}\) & \\ \hline \multirow{3}{*}{Total area (\(\mu\)m\({}^{2}\))} & BC/SC & \(3107\pm 3.112\)\({}^{\text{a}}\) & \\ \cline{2-3}  & BC+AgNPs/SC & \(5803\pm 2.479\)\({}^{\text{c}}\) & \\ \cline{2-3}  & BC/CWC & \(2546\pm 2.992\)\({}^{\text{a}}\) & \\ \cline{2-3}  & BC+AgNPs/CWC & \(4285\pm 3.153\)\({}^{\text{b}}\) & \\ \hline \hline \end{tabular} SD, standard deviation; BC, biocearamic sealer; AgNPs, silver nanoparticles; SC, single-cone technique; CWC, continuous wave condensation. For each parameter, different superscript letters indicate statistically significant differences among the groups.  
\end{table}
Table 1: Detntinal tubule penetration values of the experimental groups.