Comparison of spectroscopic and molecular methods for estimating age and sex for medico-legal purposes: protocol for a systematic review

Authors

DOI:

https://doi.org/10.62741/ahrj.v3iSuppl.230

Keywords:

Forensic Dentistry, Spectroscopy, Molecular Methods, Age estimation, Sex estimation, Dental analysis

Abstract

Introduction: In forensic contexts, accurate estimation of age and sex is fundamental for the identification of human remains. Spectroscopic methods, offering non-destructive and precise data insights, have emerged as promising alternatives/complements to traditional morphological and genetic techniques.

Objective: This protocol outlines a systematic review aimed at evaluating the effectiveness of spectroscopic and molecular methods for estimating age and/or sex based on dental samples in medico-legal settings.

Methodology: This review will be conducted in accordance with the PRISMA 2020 guidelines. Databases to be searched include PubMed, Web of Science, ScienceDirect, LILACS (via BVS), Cochrane Library (Protocols), MEDLINE (via EBSCOhost), Academic Search Complete (via EBSCOhost), CINAHL (via EBSCOhost) and grey literature. The eligibility criteria follow the PICOS framework. Risk of bias will be assessed using the QUIN tool for in vitro studies. Data synthesis will involve both quantitative and qualitative approaches. The protocol of this review has been registered in PROSPERO (CRD420250650858).

Results: This review is expected to identify a range of spectroscopic techniques applied to dental samples for age and sex estimation, including mass spectrometry, Raman spectroscopy, FTIR spectroscopy, and X-ray fluorescence, as well as molecular methods such as amelogenin analysis, DNA methylation, and telomere length assessment. The evidence is anticipated to vary regarding study design, sample characteristics, analytical protocols, and reported outcome measures. The review will synthesize the available evidence on the accuracy, applicability, and methodological characteristics of these approaches within medico-legal contexts.

Conclusion: This systematic review will provide a comprehensive overview of the available evidence regarding the use of spectroscopic and molecular methods for age and sex estimation from dental samples. By identifying the methodologies currently employed, their reported outcomes, and existing research gaps, the review may contribute to future research directions and support evidence-based decision-making in forensic and medico-legal practice, particularly in contexts where non-destructive analytical approaches are desirable.

References

Higgins D, Austin JJ. Teeth as a source of DNA for fo-rensic identification of human remains: A review. Sci Justice. 2013;53(4):433-441. doi:10.1016/j.scijus.2013.06.001 DOI: https://doi.org/10.1016/j.scijus.2013.06.001

Gamsjaeger S, Mendelsohn R, Klaushofer K, et al. Vi-brational spectroscopic analysis of hard tissues. In In-frared and Raman Spectroscopic Imaging (eds R. Salzer and H.W. Siesler); 2014. doi:10.1002/9783527678136.ch4 DOI: https://doi.org/10.1002/9783527678136.ch4

Zamojda E, Orywal K, Mroczko B, et al. Trace elements in dental enamel can be a potential factor of advanced tooth wear. Minerals 2023; 13(1):125. doi:10.3390/min13010125 DOI: https://doi.org/10.3390/min13010125

Marcante B, Marino L, Cattaneo NE, et al. Advancing forensic human chronological age estimation: bio-chemical, genetic, and epigenetic approaches from the last 15 years: A systematic review. Int J Mol Sci. 2025;26(7):3158. doi:10.3390/ijms26073158 DOI: https://doi.org/10.3390/ijms26073158

Kumari K, Rao RS, Sarode SC, et al. Raman microspec-trometry: An alternative method of age estimation from dentin and cementum. J Clin Diagn Res. 2017;11(10):ZC11-ZC16. doi:10.7860/JCDR/2017/31161.10705 DOI: https://doi.org/10.7860/JCDR/2017/31161.10705

Pishghadam N, Esmaeilyfard R, Paknahad M. Explain-able deep learning for age and gender estimation in dental CBCT scans using attention mechanisms and multitask learning. Sci Rep. 2025;15:18070. doi:10.1038/s41598-025-03305-z DOI: https://doi.org/10.1038/s41598-025-03305-z

de Almeida CM, Dos Santos NA, Lacerda V Jr, et al. Applications of MALDI mass spectrometry in forensic science. Anal Bioanal Chem. 2024;416(24):5255-5280. doi:10.1007/s00216-024-05470-y DOI: https://doi.org/10.1007/s00216-024-05470-y

Gowland R, Stewart NA, Crowder KD, et al. Sex estima-tion of teeth at different developmental stages using dimorphic enamel peptide analysis. Am J Phys An-thropol. 2021;174:859-869. doi:10.1002/ajpa.24231 DOI: https://doi.org/10.1002/ajpa.24231

Lopes Cardoso I, Moreira MT, Dupuis C, et al. Amelog-enin-based molecular methods for sexual dimorphism identification: protocol of a scoping review. Forensic Sci. 2024;4(4):499-507. doi:10.3390/forensicsci4040033 DOI: https://doi.org/10.3390/forensicsci4040033

Guimarães MI, Moreira MT, Dupuis C, et al. Determi-nation of sexual dimorphism through molecular methods: a scoping review. Egypt J Forensic Sci. 2025;15:11. doi:10.1186/s41935-025-00432-6 DOI: https://doi.org/10.1186/s41935-025-00432-6

Zapico SC, Gauthier Q, Antevska A, et al. Identifying methylation patterns in dental pulp aging: Application to age-at-death estimation in forensic anthropology. Int J Mol Sci. 2021;22(7):3717. doi:10.3390/ijms22073717 DOI: https://doi.org/10.3390/ijms22073717

Márquez-Ruiz AB, González-Herrera L, Valenzuela A. Usefulness of telomere length in DNA from human teeth for age estimation. Int J Legal Med. 2018;132(2):353-359. doi:10.1007/s00414-017-1595-5 DOI: https://doi.org/10.1007/s00414-017-1595-5

Ouzzani M, Hammady H, Fedorowicz Z, et al. Ray-yan—a web and mobile app for systematic reviews. Syst Rev. 2016;5(1):210. doi:10.1186/s13643-016-0384-4 DOI: https://doi.org/10.1186/s13643-016-0384-4

McGowan J, Sampson M, Salzwedel DM, et al. PRESS Peer Review of Electronic Search Strategies: 2015 Guideline Statement. J Clin Epidemiol. 2016;75:40-46. doi:10.1016/j.jclinepi.2016.01.021 DOI: https://doi.org/10.1016/j.jclinepi.2016.01.021

Sheth VH, Shah NP, Jain R, et al. Development and validation of a risk-of-bias tool for assessing in vitro studies conducted in dentistry: The QUIN. J Prosthet Dent. 2024;131(6):1038-1042. doi:10.1016/j.prosdent.2022.05.019 DOI: https://doi.org/10.1016/j.prosdent.2022.05.019

Downloads

Published

15-08-2026