Evaluation of the antibacterial effect of tea tree oil on Enterococcus faecalis and biofilm in vitro

Tea tree essential oil (TTO) is extracted from the leaves of Melaleuca alternifolia by steam distillation. It is well known for its traditional medicinal uses, particularly for the treatment of bruises, insect bites, skin infections, vertigo, convulsions, toothache, and rheumatism. Earlier research has shown that TTO can effectively inhibit oral microorganisms in the root canals. Enterococcus faecalis (E. faecalis) has been considered to be associated with persistent root canal infections and root canal treatment failure. The biofilm of E. faecalis makes it more vigorous, toxic, and resistant to antibiotics.In this study, our aim was to evaluate the antimicrobial effects of TTO on planktonic E. faecalis and biofilms compared with 0.2% CHX.We explored the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC), the bacteriostatic rate by MTT assay, the antimicrobial time by time-kill assay, and the effects on cell integrity, the biomass, and bacterial activity of E. faecalis biofilms. Finally, we investigated the microstructure changes of E. faecalis biofilms using scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM).The MIC and MBC values were 0.25% and 0.5%, the bacterial inhibition rate, time-kill was dosage dependent, and TTO can effectively destroy membrane integrity. SEM CLSM images revealed that TTO could reduce bacterial aggregation, biofilm thickness and inhibited biofilm formation. The effect of TTO was the same as that of 0.2% CHX at some specific concentrations. In summary, TTO has the potential to be effective against E. faecalis infections.TTO was able to inhibit E. faecalis by destroying cell membrane, inhibiting the formation of E. faecalis biofilms, and eliminating mature formed biofilms. In this study, TTO has the potential to be further developed as a novel antibacterial drug.