Differential miRNA expression during replicative senescence of dental pulp stem cells with potential for forensic age assessment

Cellular aging is a complex process that complicates biological age estimation in forensic settings due to high variability. This study evaluated whether microRNAs (miRNAs) from human dental pulp stem cells (hDPSCs) can serve as predictive biomarkers of senescence. hDPSCs from four young donors were subjected to replicative exhaustion and stress-induced senescence, with eight miRNAs selected for analysis. Senescence was confirmed by morphological changes, SA-β-Gal activity, and p16^INK4a immunocytochemistry. RT-qPCR quantified miRNA expression, and predictive value was assessed by ROC curves and multivariable models. Senescence induction consistently resulted in distinct morphological and molecular changes, notably upregulation of miR-433-5p, miR-331-5p, miR-221-5p, and miR-328-5p, and downregulation of miR-205-5p, miR-455-5p, and miR-21-5p. While individual miRNAs showed moderate predictive ability, combining multiple miRNAs greatly improved discrimination of senescence. These results support the feasibility of miRNA panels as molecular markers of senescence in hDPSCs and provide a proof-of-concept framework for future studies on cellular aging in a dental matrix of forensic interest.