Next generation forensic techniques applied on dental tissues for the identification of human remains and unaccompanied minors

The increasing humanitarian crises, mass disasters, and global migration flows have led to a significant rise in the demand for reliable forensic methods to identify human remains and estimate the age of unaccompanied minors and undocumented adults. Conventional forensic matrices (blood, urine, hair, internal organs) are frequently unavailable in severely destroyed or decomposed bodies, hampering post-mortem interval (PMI) estimation, toxicological and genetic investigations, and biological profiling and identification. In living people, ethical restrictions on ionizing dose for forensic purpose hinder the applicability of dental radiographic methods for the age estimation, increasingly preventing a scientifically reliable age assessment, with serious legal and human rights implications particularly for minors. These critical gaps underscore the urgency of validating field-adaptable forensic approaches based on alternative matrices that remain stable under extreme conditions, and non-invasive innovative age estimation methods. This project has the clear objective to experiment and validate novel analysis techniques and forensic methods based on dental tissues (pulp and dentin), saliva, and dental calculus as innovative matrices with promising perspectives for addressing relevant ethical issues and legal and forensic demands in the fields of human identification of unknown bodies and human remains, and age estimation of unaccompanied children and undocumented migrants: 1. PMI estimation via genetic analysis of post-mortem mutations occurrence in dental pulp, exploiting in-vitro death-simulation models; 2. age estimation in living unaccompanied minors through salivary DNA methylation, and in adult human remains through dental-pulp DNA methylation, as ethically acceptable alternatives to radiographic methods; 3. individual profiling and identification of unknown bodies through genetic analysis of mineralized dental tissues and calculus; 4. toxicological and chemical analysis of dental tissues and calculus to detect chronic exposure to drugs, medicines, and toxics, informing cause of death, ancestry, and the drug-accelerating ageing. The interdisciplinary integration of dentists and forensic experts as pathologists, odontologists, genetics, bioinformatics, and toxicologists is a key feature of the project, allowing the development of predictive models that reduce bias and improve estimation accuracy beyond what single-discipline studies have achieved to date. Six Italian research units will participate, collectively covering all required competencies in all investigated forensic domains. The multidisciplinary expertise of PIs and unit members are coherent with the project scope, the unit grant access to suitable samples from living individuals and dead bodies or human remains, laboratory facilities for cross-validation of all analytical protocols, and esnsure rigorous standard definition, dissemination, and exploitation of results.