| Summary: | 碩士 === 中山醫學大學 === 口腔醫學研究所 === 95 === The habit of areca (betel nut, Areca catechu) quid chewing is widespread in Taiwan, Southeast Asia, and India. Epidemiological studies have shown that areca quid chewing increases the risk of oral cancer and oral submucous fibrosis. In addition, areca quid chewers have a higher prevalence of periodontal diseases than non-chewers. In this study, the pathological effects of safrole, a major polyphenol compound in Piper betel leaf used when quid chewing, were investigated in cultured human gingival fibroblasts (HGFs). Little is known about the cytopathological effects of safrole on human gingival fibroblasts (HGF). Hence, we established 6 primary human gingival fibroblast strains for cell migration, attachment and spreading assays with the treatment of safrole. In addition, HGFs were challenged with safrole analyzed by 3-(4,5-dimethylthiazol-z-yl)-2,5-diphenyl-tetrazolium (MTT) colorimetric assay, Western blotting, gelatin zymography and casein zymography. Our results demonstrated that safrole was cytotoxic to HGFs in a dose-dependent manner (p<0.05). Safrole also inhibited cell attachment (p<0.05). A control culture exhibited a normal fibroblast monolayer of long spindle-shaped morphology. Safrole-treated HGFs showed a rounded appearance and detachment at higher concentrations. At concentrations higher than 20 µg/ml, safrole inhibited cell spreading and migration. Safrole was found to induced extracellular signal-regulated protein kinase phosphorylated (p-ERK) in a dose dependent manner.(p<0.05) Matrix metalloproteinases-2(MMP-2) and t-plasminogen activator(t-PA) expression was up-regulated by safrole treatment with time-dependent effect compare with control groups (p<0.05). Cyclooxygenase-2(COX-2) expression was increased by safrole (p<0.05). The addition of periodontal pathogens and proinflammation cytokines significant enhanced MMP-2 and t-PA expression (p<0.05) as compare with safrole alone. Taking together, these results indicate that safrole is a cytotoxic agent to HGFs. Areca quid chewers might be more susceptible to destruction of periodontium and less responsive to a regeneration procedures during periodontal therapy.
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