In vitro Reduction in Colonization of Streptococcus mutans by Honey Beeswax Ethyl Acetate Extract

Dental plaque is composed of bacterial derived extracellular polysacharide known as glucan which is synthesized by Streptococcus mutans. Natural substances that could inhibit the plaque formation of the bacteria have a significant importance. This investigation has evaluated the honey beeswax extract effect on the Gft production, the key enzyme of S. mutans colonization factor for the first time. Methods and Materials: In this experimental study extraction of the sample conducted with ethyl acetate and methanol solutions in the Clevenger extractor. The ethyl acetate soluble fraction was separated in the first step and after the evaporation of the first solute, the 70% methanol as inactive solvent was added and the water mixture was used as a second solution, then materials were separated with dH2O. Minimum inhibitory concentration (MIC) of the honey beeswax extract was assessed by Broth diffusion method. Examination of cell adherence (Biofilm Inhibitory Concentration, BIC) was calculated by colony counts from surface scratching of glass slides in the bacterial media that supplied with 1% sucrose. Glucosyltransferase expression was detected by 15% SDS poly acrylamide gel electrophoresis. Concentration of 1mg/ml of ethyl acetate honey beeswax extract was inhibited completely biofilm and it was prevented the production of glucosyltransferase enzyme. The concentration of formation 6 mg/ml of the extract had bacteriostatic effect and 30 mg/ml concentration of this extract had bactericidal for S. mutans (P<0.01). The sub- bacterial concentrotion honey beeswax extract was able to block the major enzyme that contributes to S. mutans biofilm formation.