The antimicrobial and antibiofilm effects of three herbal extracts on Streptococcus mutans compared with Chlorhexidine 0.2% (in vitro study)

Abstract

There is a special focus on using natural materials and herbal plants to prevent dental caries. Previous studies showed that some herbal plants have antimicrobial effects on oral pathogens. Thus we investigated the antimicrobial effects of three herbal extracts (Carum copticum, Phlomis bruguieri, and Marrubium parviflorum) on the growth of Streptococcus mutans, as the most important bacteria causing dental caries. First, plant methanolic extracts were prepared. Then, to evaluate the antimicrobial activity of the three herbal extracts, the agar well diffusion method and MIC were performed. The biofilm formation was carried out using a broth dilution method with 2% glucose-supplemented BHIS in sterile 96-well microplates. Serial dilutions (50, 25, 12.5, 6.25, 3.12 mg/ml) of extracts were prepared. Next, a 0.5 McFarland Suspension of S. mutans was added to wells. The inhibitory effect on biofilm formation was measured by the ELISA reader apparatus. The assay was repeated three times, and the average was calculated as 3. The results were compared with those of Chlorhexidine 0.2%. Carum copticum showed a better effect in the agar well diffusion method than others. MIC of the extracts of Carum coptimum, Phlomis bruguieri, and Marrubium parviflorum were 3.12, 6.25, and 12.5 mg/ml, respectively. Overall, the highest activity belonged to Carum copticum extract. For the anti-biofilm effect, the OD values of Carum copticum and Marrubium parviflorum were significantly different from that of Phlomis bruguieri. Although all of the methanolic herbal extracts can inhibit S. mutans growth and remove the biofilm, the effect of Carum copticum was better than Phlomis bruguieri and Marrubium parviflorum. Further studies are recommended to indicate how these extracts perform against the bacteria.

Keywords: ATP – Adenosine triphosphate; BHI – Brain-heart infusion broth; BHIS – Brain Heart Infusion broth plus sucrose 2%; Carum copticum; DMSO – Dimethyl Sulfoxide; MDR – Multi drug resistance; MIC – Minimum Inhibitory Concentration; Marrubium parviflorum; OD – Optical Density; PBS – Peripheral Blood Smear; Phlomis bruguieri; Streptococcus mutans.