Cytotoxicity and Morphological Effects of Aqueous Areca Nut Crude Extract on L929 Fibroblast Cell Line

Abstract

Betel quid chewing is a detrimental recreational habit amongst Asians and a risk factor for oral cancer. Arecoline, a component of areca nut (a major constituent of betel quid) is a known carcinogen. However, the effect of areca nut crude extract is not much studied. To evaluate the cytotoxicity and morphologic effects of areca nut aqueous extract on mouse fibroblast cell line (L929). Dried raw areca nut obtained from a local market in Kota Bharu, Kelantan was prepared and suspended in DMEM (Dulbecco’s Modified Eagle’s medium), prior to serial dilution of 1.56, 0.781, 0.39, 0.195, 0.0976, 0.0488, and 0.0244 mg/ml. The L929 was then exposed to each of the aqueous areca nut extract dilutions and incubated at 37 °C for 24, 48 and 72 hours. Following incubation, the cytotoxicity level of treated L929 was measured using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium] assay. Five highest concentrations of areca nut extract showed significantly decreased L929 cell viability (1.56, 0.781, 0.39, 0.195, 0.0976 mg/ml) for all incubation periods compared to untreated cells (p<0.05). The IC50 values of aqueous areca nut extract on L929 were 0.1516, 0.1040, and 0.09136 mg/ml at 24, 48 and 72 hours, respectively. The L929 cell showed altered morphology when cultured in the extract for 24 hours. Higher concentrations of the areca nut aqueous extract is cytotoxic to L929. Prolonged exposure to the extract reduced the IC50 value. Investigation on the role of areca nut in cell proliferation could be further undertaken to assess its association with oral cancer.
 
 
 
 Keywords: areca nut, L929, mouse fibroblast cell line, cytotoxicity, MTT assay