Epithelial-mesenchymal interaction reduces inhibitory effects of fluoride on proliferation and enamel matrix expression in dental epithelial cells

AIM: Fluoride, well known as a specific and effective caries prophylactic agent, also affects the differentiation and function of ameloblasts. High dose sodium fluoride (NaF) induces enamel hypoplasia, also called enamel fluorosis, whereas the size and form of teeth except the enamel are not changed with its treatment. We examined the effects of fluoride on dental epithelium proliferation and differentiation using co-cultures of dental epithelial and mesenchymal cells. METHODS: Cultures of the dental epithelial cell line SF2 and dental mesenchymal cell line mDP were performed, as well as co-cultures. Enamel matrix expression in SF2 cells treated with NaF was analyzed by RT-PCR, while cell proliferation was examined using a trypan blue dye exclusion method and BrdU incorporation findings. The effects of NaF on NT-4-induced ERK1/2 phosphorylation were analyzed by western immunoblotting. RESULTS: Neurotrophic factor NT-4 induced enamel matrix expression, which was inhibited in the presence of NaF. Similar results were observed in regard to SF2 cell proliferation, but not with mDP cells. The levels of proliferation and ameloblastin expression in SF2-GFP cells co-cultured with mDP in the presence of NaF were lower as compared to those in SF2 cells cultured alone. CONCLUSION: Our results indicate that dental epithelial cells co-cultured with dental mesenchymal cells are resistant to the inhibitory effects of NaF on proliferation and ameloblastin expression. They also suggest that the dental fluorosis phenotype may affect enamel, but not tooth size or shape, because of rescue of the inhibitory effects of NaF by culturing with dental mesenchymal cells.Key words Amelogenesis, Enamel, Enamel matrix, Epithelial-mesenchymal interaction, Fluoride.