Increased fractal complexity of the epithelial-connective tissue interface in the tongue of 4NQO-treated rats

The effect of the carcinogen 4-nitroquinoline 1-oxide (0.001% in drinking water) on the irregularity of the epithelial connective tissue interface (ECTI) of the ventral surface of the tongue was quantified in Dark Agouti and Wistar Furth rats. Histological tongue sections stained with the Azan-Mallory method were digitised (111 images, resolution 1 pixel = 3.1 μm), and the limit between epithelium and stroma of the ventral surface was extracted and analysed using a fractal geometry technique (local connected dimension). The results showed that although none of the images included carcinomas on the ventral surface of the tongue (all cases had other oral carcinomas), the epithelial profiles of the treated cases showed a statistically significant increase in irregularity when compared to controls. Canonical discriminant analysis of the parameters describing the irregularity of the ECTI classified 81.1% of the images in the original groups (treated or control). Fractal analysis is capable of detecting subtle architectural changes in the oral epithelium of the rat occurring after exposure to the carcinogen, even when full malignant transformation has not yet taken place. Fractal analysis, which may prove useful for monitoring the progression of carcinogenesis in this animal model, is a morphometrical parameter in the diagnosis of oral epithelial dysplasia.