The age threshold of 14 years is relevant in Italy as the minimum age for criminal responsibility. It is of utmost importance to evaluate the diagnostic accuracy of every odontological method for age evaluation considering the sensitivity, or the ability to estimate the true positive cases, and the specificity, or the ability to estimate the true negative cases. The research aims to compare the specificity and sensitivity of four commonly adopted methods of dental age estimation – Demirjian, Haavikko, Willems and Cameriere – in a sample of Italian children aged between 11 and 16 years, with an age threshold of 14 years, using receiver operating characteristic curves and the area under the curve (AUC). In addition, new decision criteria are developed to increase the accuracy of the methods. Among the four odontological methods for age estimation adopted in the research, the Cameriere method showed the highest AUC in both female and male cohorts. The Cameriere method shows a high degree of accuracy at the age threshold of 14 years. To adopt the Cameriere method to estimate the 14-year age threshold more accurately, however, it is suggested – according to the Youden index – that the decision criterion be set at the lower value of 12.928 for females and 13.258 years for males, obtaining a sensitivity of 85% and specificity of 88% in females, and a sensitivity of 77% and specificity of 92% in males. If a specificity level >90% is needed, the cut-off point should be set at 12.959 years (82% sensitivity) for females.

Teeth may serve as alternative matrices for post-mortem toxicological analysis. Although the mechanism of drug incorporation into dental tissues remains unclear, it is hypothesized that orally ingested substances can permeate dental enamel and reach dentin and the pulp. This study investigated enamel permeability to commonly ingested drugs, considering molecular characteristics (methadone - MET and dextromethorphan hydrobromide - DXM), pH, sugar content, and tooth conditions (intact enamel, cement-enamel junction-CEJ exposure, enamel wear). Sixteen clinically extracted teeth were divided into three groups and immersed in different substances, including MET with sugar, DXM with and without sugar (pH 4.5, 1 mg/ml), and acidified saliva (pH 5-6), simulating an addicted individual's oral environment. Cyclic immersions (substance-saliva-substance) mimicked chronic drug intake for about six weeks. After treatment, each tooth was separated into enamel, dentin, and pulp, and analyzed using liquid-liquid extraction and LC-MS/MS. Preliminary results showed that both MET and DXM can penetrate both the enamel and dentin, reaching the pulp regardless of tooth integrity or molecule type. Acidity and sugar content significantly increased the diffusion. These findings suggest that other substances orally assumed might be endowed with similar effects on dental tissues, especially when combined with sugar or acid solutions, supporting teeth as viable matrices in post-mortem toxicology. However, pulp concentrations may reflect oral contamination rather than systemic levels.