In vitro evaluation of the cytotoxicity of chlorhexidine digluconate, povidone iodine and phenolic compound mouth washes on chinese hamster v79 cell lines

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Aim: To assess and compare the cytotoxic potential of Chlorhexidine, Listerine and Povidone –Iodine antimicrobial agents on Chinese hamster lung fibroblast V79 cells. Materials and Method: Chlorhexidine (0.2%), Listerine, Povidone iodine were diluted with DMEM medium under sterile conditions to obtain the required concentration. V79 cells were used for the experimental procedures. The cells were cultured in DMEM media supplemented with 10% FBS and 1% Gentamycin at 370 C in a humidified 5% CO2 incubator. For all the assays, cells were maintained at 80-90% confluency in T-25 cm2 flasks and used as per requirement. V79 cells were seeded onto 96 well plates at a density of 104 cells per well and allowed to incubate overnight at 37 0 C in 5% CO2 incubator. The following day the cells were treated with different concentrations of Betadine, Hexidine and Listerine for 12, 24 and 48 hours. Following the treatments, the drug containing media was discarded and 100µl of freshly prepared MTT stock prepared in DMEM media was added in to each well and the plates were incubated at 370 C in 5% CO2 incubator for 4 hours. Viability was determined by the ability of the cells to reduce MTT, a water soluble tetrazolium dye to produce violet crystals of formazan. Formazan crystals formed were solubilized by the addition of 100µl of (Di Methyl Sulfoxide). Optical density which is directly proportional to cell viability, was measured at 540nm using a multi well spectrophotometer. Viability curves were plotted using origin. Percentage cell viability was obtained using the formula given below: Percentage viability= [(OD Test – OD Blank)/ (OD Control – OD Blank)] *100 Results: V79 cells treated with serially diluted test compounds showed a decrease in cellular viability in a concentration as well as time dependent manner. Hexidine exhibited maximum toxic effect when compared to Listerine and Betadine; betadine possessing the least toxic nature as obvious by the IC50 values for different time durations. Conclusion: Chlorhexidine was found to be most toxic when compared to Listerine and Povidone iodine. Povidone iodine being the least toxic amongst the three mouth rinse. All the compounds exhibited toxic effect in a dose dependent as well as time dependent manner. The result of this study showed that all three given mouth rinses caused V79 proliferation inhibition.