In vitro evaluation of cytoproliferative activity of areca nut varieties on murine fibroblasts.

INTRODUCTION

Areca nut, derived from the Areca catechu palm, is consumed by 10-20% of the global population and is linked to oral submucous fibrosis (OSMF), a potentially malignant oral disorder marked by increased fibroblast proliferation. The processing of areca nut involves husking, nut removal, drying, roasting, boiling, and fermentation.

AIM: This study aims to examine how various processing methods and storage conditions of areca nut affect L929 fibroblastic cell lines.

MATERIALS AND

METHODS

Five areca nut varieties which underwent different processing methods and five commercial areca nut products (without tobacco) from Karnataka, Kerala, and Gujarat were collected and extracted using a triple solvent solution. Cytoproliferative activity was assessed using the methyl thiazolyl tetrazolium (MTT) and Trypan blue dye exclusion assays. Varieties with the highest proliferative activity were further analysed by Reverse transcription-Polymerase chain reaction (RT-PCR) and immunofluorescence, targeting collagen, fibronectin, and α-SMA expression.

RESULTS

Certain areca nut extracts stimulated fibroblast proliferation. However, there were no significant differences across processing methods, storage conditions, or chemical treatments. Two areca nut varieties and one commercial product exhibited the highest expression of collagen, fibronectin, and α-SMA, confirmed by both RT-PCR and immunofluorescence.

CONCLUSION

The elevated expression of fibrosis-related proteins in two areca nut varieties with differing processing and storage suggests a potential role in fibrosis development. Only one of five commercial products showed similar activity, indicating variability in commercial preparations. These findings underscore the importance of further research into how processing and storage influence areca nut's role in OSMF.