Cytotoxicity evaluation of effective eco produce as endodontic irrigant: an in vitro study

Abstract

Sodium hypochlorite (NaOCl), the gold standard for root canal irrigation, carries the risk of potential adverse effects. Effective Eco Produce (EEP), an organic solution produced from pineapple and orange peels is shown to exhibit antibacterial and antibiofilm activities, demonstrating its potential as an alternative endodontic irrigant. However, there are limited studies on the cytotoxicity of EEP. This research aimed to evaluate the cytotoxic effect of EEP at different concentrations and fermentation periods towards MC3T3-E1 pre-osteoblast cell line in vitro. EEP was prepared from a mixture of pineapple and orange peel waste, which were fermented for three and six months. MC3T3-E1 pre-osteoblast cells were treated with eight concentrations EEP at both fermentation periods with 2-fold serial dilutions, ranging from 100% to 0.78% for 24 hours. The 2.5% NaOCl group was used as the comparison group. The cell viability was analysed using the Alamar Blue and Live and Dead Cell assay. Ultrastructural morphological changes of the cells were evaluated using transmission electron microscope (TEM). Statistical analysis of the data was performed using twoway mixed Analysis of Variance (ANOVA). The cytotoxic effect of EEP was concentration dependent. EEP at a concentration below 6.25% was considered not cytotoxic with relative cell viability of more than 70% and comparable to the negative control (p > 0.05). Different fermentation periods have no statistically significant effect on cell viability (p > 0.05). There was a significantly higher cell viability among all concentrations of EEP when compared to the sodium hypochlorite group at both three and six months of fermentation (p < 0.05). Fluorescence microscopic visualisation of Live and Dead Cell assay complemented the findings. TEM analysis of the 6.25% EEP for both fermentation periods was comparable to negative control with intact cell membranes and organelles. The same research team showed that 6.25% EEP (6-month fermentation) exhibited a high anti-biofilm effect. Therefore, 6.25% EEP at 6 months of fermentation shows potential as a safer alternative to 2.5% NaOCl to be used as endodontic irrigant.