New protocols for studying the biological and paleobiological materials and for assessing diagenetic processes

This report is part of the work performed in the project PNRR PE 5 “CHANGES”, and specifically in Spoke 5 “Science and Technologies for Sustainable Diagnostics for Cultural Heritage”, WP4 “Advanced diagnostics and scientific methods for organic material”It presents and discusses the results obtained in the project, planned as: R4.1 Development and implementation of digital, AI-based methods, chemical/physical and molecular approaches for studying the biological and palaeobiological materials and constitutes one of the Deliverables produced: D4.1 – New protocols for studying the biological and paleobiological materials and for assessing diagenetic processes.The scope of this report is to present and discuss the activities performed in WP4 by the research partners involved. The UNIBO Cultural Heritage Department integrated analytical study of dental mineralized tissues offers a powerful framework for reconstructing life histories in an archaeological population, yielding information that is rarely accessible from other skeletal elements. Histological investigation of dental enamel, combining classical histomorphometry with histochemical analysis via LA-ICP-MS, offers the opportunity to examine the behaviour of enamel-forming cells during early-life stress episodes. This dual approach provides a more refined interpretation of the potential causes and developmental timing of enamel disturbances. Furthermore, coupling histomorphometric data with computer-based analytical algorithms present a promising method for estimating age-at-death in adults through dental cementum analysis. Since accurate age estimation in adults remains one of the most challenging aspects of biological profile reconstructions, this innovative approach has the potential to substantially improve both precision and reliability for this purpose. The overarching goal of the UNIBO Chemistry Department is to develop rapid, reliable, and minimally destructive analytical approaches for archaeological biomaterials. This includes: improving the characterization of burned bones through portable IR and NIR spectroscopy coupled with chemometric analysis; assessing cellulose preservation in archaeological wood using SWIR hyperspectral imaging to optimize sampling for radiocarbon dating; evaluating long-term amino acid preservation in fossil enamel as a proxy for intra-crystalline organic retention; and advancing fast, low-cost detection of amelogenins through innovative biosensor platforms. Together, these activities aim to enhance diagnostic capabilities, refine sampling strategies, and broaden the analytical toolkit available for archaeological research. The UNIFI (BIO) project aims to shed new light on the lives of the people of Pompeii and on other very important samples ( i.e Grotta delle Mura individual) by combining the most advanced methods in genomics, bioarchaeology and biochemical analysis. Thanks to modern palaeogenomic techniques, researchers can now extract and study DNA preserved in ancient human remains, even when the material is highly degraded. By applying these methods to the skeletal remains and dental calculus of more than 170 Pompeii inhabitants, the studi seeks to reconstruct not only their biological characteristics—such as sex, physical appearance, genetic traits, intolerances and diseases—but also their family relationships and patterns of movement or migration. Stable isotope analyses of carbon, nitrogen, sulfur and strontium will offer further insight into what these individuals ate and whether they had moved during their lifetime. All these biological findings will be interpreted alongside archaeological, historical and cultural evidence to build a richer and more nuanced picture of daily life in the Roman city before the eruption of Vesuvius in 79 CE. Early tests indicate that the DNA is well preserved, making it possible to generate high-quality genomic data using state-of-the-art sequencing technologies. By comparing Pompeii’s genomes with vast datasets of worldwide human genetic diversity, researchers will be able to trace the deeper ancestry and long-term connections of the population, extending from prehistory to the present day. Ultimately, the project hopes to make significant contributions to our understanding of Roman society, offering new knowledge not only to scholars but also to museum visitors, documentary producers and the general public. Its emphasis on the preservation and enhancement of cultural heritage promises meaningful cultural, scientific and social benefits at local, regional and national levels. The experiences and the new protocols reported contribute to the needs of supporting the knowledge consolidation process with digital instruments and scientific methods, leading to the common goal of a better diagnostic investigation of organic material, and in particular for studying the biological and paleobiological materials and for assessing diagenetic processes.