Evaluation of the antibacterial activity of cinnamon essential oil and its individual compounds on Aggregatibacter actinomycetemcomitans isolated from black extrinsic tooth stain: an in vitro study.

AIM: Black extrinsic tooth stain (BETS) is a health challenge that commonly affects children. Aggregatibacter actinomycetemcomitans (Aa) presents in higher prevalence within the polymicrobial community of BETS. In this study, the anti-planktonic and anti-sessile activities of cinnamon essential oil (CEO) and its individual compounds against Aa were evaluated. The preventive effect of CEO and its active substances on BETS formation was also studied in vitro. METHODS: Aa was isolated from a preschool child with BETS and was identified based on the morphological characteristics, MALDI-TOF mass spectroscopy and 16S rRNA sequencing. The effect of CEO and its individual compounds on the growth kinetics of planktonic and sessile Aa cells as well as their antibacterial efficacy and their rate of bacterial killing were examined. The preventive effect of CEO and its active substances on the formation of BETS was evaluated using an ex vivo model. The data were analysed using one-way analysis of variance (ANOVA) and the significance level was set at p < 0.05. RESULTS: Out of eight individual compounds of CEO, only eugenol, cinnamaldehyde and α-methyl cinnamaldehyde showed anti-Aa activities. The values of the minimum inhibitory concentrations (MICs) were in the following order: CEO (421.5 mg/ml) > α-methyl cinnamaldehyde (26.37 mg/ml) > cinnamaldehyde (0.209 mg/ml) > eugenol (0.052 mg/ml). CEO, eugenol, cinnamaldehyde and α-methyl cinnamaldehyde, respectively, exhibited two-, four-, four- and eightfold increase of sessile MIC compared to their planktonic MIC. The growth kinetics of both planktonic and sessile Aa in the presence of CEO, eugenol, cinnamaldehyde and α-methyl cinnamaldehyde revealed a complete inhibition at the MICs and 5.3%-37.4% biofilm inhibition at sub-MICs. The time-killing study demonstrated that CEO, eugenol and cinnamaldehyde were capable of reducing the survival rate of both planktonic and sessile Aa cells after 15-20 and 25-30 min, respectively. However, α-methyl cinnamaldehyde showed a superior anti-planktonic to anti-biofilm activity. The daily incorporation of CEO, eugenol and cinnamaldehyde at their MICs for 14 days totally prevented the formation of BETS in the ex vivo model; however, in the case of α-methyl cinnamaldehyde, BETS was visually detectable after 10 days. CONCLUSION: CEO and its individual compounds have marked antibacterial activity against Aa. The effective results against planktonic and sessile Aa within reasonable time indicate that they can be used to prevent BETS.

Evaluation of the antibacterial activity of Enamelast® and Fluor defender® fluoride varnishes against Streptococcus mutans biofilm: an in vitro study in primary teeth.

PURPOSE: The aim of the current work was to compare the antibacterial activity of Enamelast® and Fluor defender® fluoride varnish on biofilm generation by Streptococcus mutans on extracted primary teeth. METHODS: Thirty-six primary molars were collected and sliced into seventy-two test model disks. All specimens were examined, and the cracked or broken ones were discarded. A total number of specimens (n = 54) were divided into two experimental analyses viz; biofilm formation (n = 27) and microscopic examination (n = 27). Specimens of each analysis were tested under different experimental conditions: a negative control group (n = 9), Fluor defender group (n = 9), and Enamelast group (n = 9). Following treatment, biofilms were generated by adherent Streptococcus mutans on the test model disks on three time intervals: 24 h (n = 3), 48 h (n = 3), and 72 h (n = 3) for each analysis. Then, for biofilm formation analysis, the biofilm was detected spectrophotometrically at 620 nm after being stained by crystal violet. For microscopical analysis, the surfaces of the test model disks were visualized by scanning electron microscopy (SEM), and each image was processed and analyzed using ImageJ software. RESULTS: At 48 and 72 h, Enamelast® and Fluor defender®-treated group showed significantly (p < 0.001) slight adhered bacterial cells when compared with the negative control group as revealed by the absorbance and SEM. Compared with the Fluor defender®-treated group, the absorbance of the Enamelast®-treated group showed a significant (p < 0.001) increase by approximately 7- and 16.5-fold at 48 and 72 h, respectively. Similarly, SEM showed that the number of bacterial cells adhered to enamel surfaces in the Fluor defender®-treated group was significantly (p < 0.001) fewer than the Enamelast®-treated group by approximately 36.55% and 20.62% at 48 and 72 h after exposure, respectively. CONCLUSION: We conclude that the anti-biofilm activity of Fluor defender® against Streptococcus mutans was significantly (p < 0.001) greater than Enamelast® fluoride varnish. The use of Fluor defender® is encouraged as a preventive measure in children with the high risk of developing dental caries.