Age estimation is vital in forensic odontology, with exfoliative cytology and cytomorphometry emerging as promising non-invasive techniques. This study evaluates the accuracy of cytomorphometric parameters like nuclear perimeter (NP), cell perimeter (CP), and NP:CP ratio, in estimating age and gender across various age groups.

This cross-sectional study involved 120 participants (60 males and 60 females) divided into six age groups (10–20, 21–30, 31–40, 41–50, 51–60, ≥ 61 years). Exfoliative cytology was performed to collect buccal mucosal cells, stained with Papanicolaou stain and analyzed for NP, CP, and NP:CP ratio in 50 cells per smear at 400 × magnification. Statistical analysis was conducted.

A significant decline in mean NP and CP values was observed with increasing age, while NP:CP ratio showed a slight increase in those aged ≥ 61 years. On intergroup comparisons, notable differences were seen between juvenile/early adulthood and middle/older adulthood groups. Regression analysis showed CP had the highest age prediction accuracy (31.7%), with a combined model improving accuracy to only 35.8%. Logistic regression using NP and CP achieved 90% predictive accuracy in distinguishing juvenile from older adulthood, with CP as the primary predictor. No significant gender differences were found, except for NCR, which was higher in females.

This study demonstrates that the cytomorphometric evaluation (NP, CP, NP:CP ratio) of exfoliated buccal mucosal cells is not a potent tool for age estimation due to its moderate predictive accuracy. While it excels in distinguishing between distinctly separated age groups, its precision falters in closely adjacent groups, rendering it of limited forensic relevance.