Antibacterial and anti-biofilm activities of chinese Propolis essential oil microemulsion against Streptococcus mutans.

AIMS: To solve the shortcomings of poor solubility, easy volatilization and decomposition, propolis essential oil microemulsion (PEOME) was prepared. The antibacterial, anti-biofilm activities and action mechanism of PEOME against Streptococcus mutans was analyzed.

METHODS: PEOME was prepared using anhydrous ethanol and Tween-80 as the cosurfactant and surfactant respectively. The antibacterial activity of PEOME against Streptococcus mutans was evaluated using the agar disk-diffusion method and broth microdilution method. The effects of PEOME on S. mutans biofilm was detected through the assays of crystal violet (CV), XTT reduction, lactic dehydrogenase (LDH) and calcium ions leaking, live/dead staining and scanning electron microscopy (SEM). And the anti-biofilm mechanism of PEOME was elaborated by the assays of extracellular polysaccharide (EPS) production and glucosyltransferase (GTF) activity.

RESULTS: The inhibition zone diameter (DIZ) of PEOME against S. mutans was 31 mm, while the minimal inhibitory concentration (MIC) was 2.5 µL mL-1. CV and XTT assays showed that PEOME could prevent fresh biofilm formation and disrupt preformed biofilm through decreasing the activities and biomass of biofilm. The leaking assays for LDH and calcium ions, as well as the live/dead staining assay, indicated that PEOME was able to damage the integrity of bacterial cell membranes within the biofilm. SEM revealed that PEOME had a noticeable inhibitory effect on bacterial adhesion and aggregation through observing the overall structure of biofilm. The assays of EPS production and GTF activity suggested that PEOME could reduce EPS production by inhibiting the activity of GTFs, thus showing an anti-biofilm effect.

CONCLUSIONS: The significant antibacterial and anti-biofilm activities against S. mutans of PEOME meant that PEOME has great potential to be developed as a drug to prevent and cure dental caries caused by S. mutans.