Determination of phenotypic alteration of arecoline-induced buccal mucosal fibroblasts: An in-vitro cell culture study

Abstract

Introduction: Oral cancer is one of the worldwide health problems accounting as the 6th common of all malignancies. Majority of the oral cancer develop from premalignant conditions of the oral cavity due to the chronic habit of tobacco chewing and smoking. The prominent cells of the oral mucosa are fibroblasts playing a major role in synthesis of extracellular matrix, wound healing, and wound repair. Arecoline, one of main ingredients of tobacco is considered as a risk factor for the development of oral premalignant lesions and cancer. The arecoline is reported to have both genotoxic and morphological alteration of oral fibroblasts leading to Oral submucous fibrosis. Thus in our study a dose dependent effect of arecoline was assessed on the morphology of cultured Human Buccal Mucosal Fibroblasts. Aim: The aim of our experiment is to assess the effect of different concentration of arecoline on the morphological variation of primary cell lines of human buccal mucosal fibroblasts to develop a model of altered fibroblasts. Materials and Methods: The primary cell lines of human buccal mucosa were established in BSRC KAHER Belagavi, and authenticated by STR profiling from DNA forensics Lab New Delhi. The cells were further cultured and assessed after treating with different concentration of arecoline hydrobromide. The treated cells were then observed for the phenotypic changes and recorded. The morphological alterations were compared to the untreated fibroblasts. Results and Conclusion: In our study, a dose-dependent effect of arecoline was assessed on phenotypic or morphological alteration of buccal mucosal fibroblasts. The results justified that concentration of arecoline lower than 125 μg/ml did not show change in the morphology of buccal mucosal fibroblasts, whereas the concentration of arecoline >250 μg/ml showed altered fibroblasts. Hence, it can be concluded that the levels of arecoline in arecanut chewers if it is >250 μg/ml, the mucosal fibroblasts may undergo changes to cause fibrosis of collagen. The future scope of our study is to determine the genotoxic effects of arecoline on buccal mucosal fibroblasts and also to develop the therapeutic effects.

Keywords

• arecoline hydrobromide
• cytotoxicity
• human buccal mucosal fibroblasts
• oral submucous fibrosis