Cytotoxicity was judged using tetrazolium bromide reduction assay. Cell migration was evaluated by Transwell assay. In vitro mineral nodule formation was assayed by von Kossa staining. Cell differentiation was examined by alkaline phosphatase activity with substrate assay. The production of osteoprotegerin was evaluated using enzyme-linked immunosorbent assay.
Arecoline demonstrated cytotoxicity to cementoblasts in a dose-dependent and time-dependent manner (P < 0.05). Arecoline attenuated cell migration in a dose-dependent manner (P < 0.05). Arecoline treatment markedly suppressed cementoblast-mediated biomineralization in vitro compared to untreated cells at Day 8. Arecoline was found to inhibit alkaline phosphatase activity in a time-dependent manner (P < 0.05). In addition, arecoline decreased the secretion of osteoprotegerin in a dose-dependent manner (P < 0.05).
Taken together, these results suggest that arecoline could inhibit cell growth, migration, and differentiation in cementoblasts. Areca quid chewers might be more susceptible to the destruction of periodontium and less responsive to regenerative procedure during periodontal therapy.