Effects of snuff extract on epithelial growth and differentiation in vitro.

Snuff is a locally irritative agent causing hyperkeratinization and hyperplasia of the oral epithelium. The aim of the study was to investigate the effects of snuff on epithelial cell growth and differentiation in vitro. Three-dimensional HaCaT cell cultures were grown for 6, 12, 14, and 18 days in the presence of 1% snuff extract. Ki-67, p53 and cytokeratins (Cks) 5, 13, 10, 19, 18, involucrin and filaggrin were studied by means of immunohistochemistry. Ki-67 indices were assessed, and the results analyzed statistically. Marked morphologic changes were seen with advanced culture time in the snuff group, probably as a result of increased toxic effects. Snuff exposure decreased the percentage of Ki-67 positive cells on days 6, 12, and 14, suggesting that snuff does not stimulate proliferation activity in this in vitro model. Cornification-related Ck 10 decreased after snuff exposure, indicating disturbances in the epithelial differentiation process.