## 4 Discussion  
The present study showed that biochemical markers, such as TNF-\(\alpha\), SOD-3, osteocalcin, and TGF-\(\beta\)1, have statistically significant higher levels in acute and chronically inflamed versus healthy pulp samples obtained from primary teeth. Moreover, osteocalcin presented statistically significant elevated levels in the chronically inflamed pulp versus acute inflamed pulp tissues (Tables 1 and 2, Figures 1-4). All biomarkers were detected in the three groups of vital dental pulp specimens. The study hypothesis was confirmed as the levels of the biomarkers differed in all three groups of pulp samples. The studied biomarkers have strong interconnections in the inflammation processes of the dental pulp characterized by molecular and cellular mechanisms. The results of the present research are analogous to the ones of our previous study, in which we evaluated the levels of TNF-\(\alpha\), SOD-3, osteoclastcin, and TGF-\(\beta\)1 in symptomatic irreversibly inflamed pulp samples obtained from the permanent teeth of children and adolescents. In both studies, symptomatic irreversibly (acute) inflamed pulp tissues revealed significant elevated levels of the studied biomarkers compared to the control group. These results may indicate the existence of similar mechanisms of inflammation, oxidative stress, and mineralization in the primary and permanent teeth of children and \begin{table}
\begin{tabular}{c c c c} \hline
**Comparison/Variable** & **Healthy vs. Acute** & **Healthy vs. Chronic** & **Acute vs. Chronic** \\ \hline TNF-\(\alpha\) * & \textless{}0.001 & 0.001 & 1.000 \\ \hline SOD-3 ** & \textless{}0.001 & \textless{}0.001 & 1.000 \\ \hline Osteocalcin ** & 0.006 & \textless{}0.001 & 0.002 \\ \hline TGF-\(\beta\)1 ** & 0.001 & 0.014 & 0.082 \\ \hline \end{tabular}
* [PERSON]–[PERSON] post-hoc test, ** [PERSON]–[PERSON] post-hoc test.  
\end{table} 
Table 2: Post-hoc analysis of protein expression of biomarkers in the control and experimental groups. Figure 4: TGF-\(\beta\)1 protein levels (ng/mg) in the control group versus acute/chronic pulpitis. (* Indicates statistically significant differences). (The circle in the graphic is an indication that an outlier is present.) Figure 3: Osteocalcin protein levels (ng/mg) in the control group versus acute/chronic pulpitis. (* Indicates statistically significant differences). The circles in the graphic are an indication that an outlier is present. )adolescents, although, from a clinical and histological perspective, particularities exist (see introduction). Regarding the levels of the studied biomarkers--in chronically inflamed versus healthy pulp samples obtained from primary teeth, all of them expressed statistically significant higher levels in the experimental compared to the control group. Statistically significant elevated levels of other biological markers, e.g., IL-4 and IgE, were also found in chronically inflamed pulp samples, such as in the study by [PERSON] et al. [19], in which the experimental group precisely consisted of chronic hyperplastic pulpitis from the permanent teeth of young patients. [PERSON] et al. [2] determined the protein expression of IL-1\(\beta\) in chronic open pulpitis, chronic closed pulpitis, and healthy pulp samples, using ELISA, without data about the provenience of the pulp tissues. IL-1\(\beta\) levels were found to be statistically significantly higher in the group with chronic closed pulpitis compared to the chronic open pulpitis (\(p<0.01\)), as well as in the group with chronic closed pulpitis versus the controls (\(p<0.01\)). They did not report any statistically significant differences between chronic open pulpitis and the control group (\(p=0.081\)). The differences in the overall concentration of IL-1\(\beta\) in chronically inflamed versus healthy pulp tissues was not statistically significant (\(p=0.590\)). The results of a study conducted by [PERSON] et al.