IADR Abstract Archives
Probiotic Lactobacillus Reuteri Eliminates Mixed-Species Endodontic Biofilms Through Reuterin-Based Mechanism
Objectives: Biofilm infection is the major cause of treatment failure of the endodontically-treated teeth. Enterococcus faecalis and Fusobacterium nucleatum are two major endodontic pathogens responsible for biofilm infections. The aim of the present study was to investigate the ability of probiotic bacteria Lactobacillus reuteri to eliminate mixed-species endodontic biofilms of foregoing endodontic pathogens when used as an irrigating solution.
Methods: Firstly in vitro activity of planktonic, mono-species and mixed-species biofilms of the Lactobacillus reuteri was determined using MIC assays in 96-well plates. Thereafter, a total of 60 human premolar teeth samples were collected, prepared, and sterilized for ex vivo experiments. All teeth samples were inoculated with mono-species and mixed-species of Enterococcus faecalis and Fusobacterium nucleatum and cultured for 24 hours for biofilm formation. Teeth were divided into 3 groups according to the irrigant type. group 1 was 2.5% NaOCl (positive control), group 2 was saline (negative control) and, group 3 was the experimental reuterin isolate irrigant. The endodontic samples were obtained at 5 minutes and 30 minutes post-irrigation to determine the number of E. faecalis and F. nucleatum. In addition, DNA was extracted and individual bacterial counts were evaluated using Real-time PCR. Data were statistically analyzed via one-way ANOVA, where p<0.05 was set as the level of significance.
Results: Reuterin isolated from L. reuteri showed promising antibiofilm activity. The antibiofilm concentration of reuterin against biofilms of E.faecalis and F.nucleatum biofilms was significant compared to the negative control (p< 0.05) and similar to that of NaOCl positive control. Interestingly, reuterin also demonstrated anti-biofilm activity in ex vivo tooth model at the concentration of 25µg/mL against E. faecalis and F. nucleatum.
Conclusions: The present study has for the first-time demonstrated that reuterin isolate Lactobacillus reuteri can successfully eliminate pathogenic endodontic biofilms as root canal irrigant. This new finding will lead to clinical benefits in future.
Methods: Firstly in vitro activity of planktonic, mono-species and mixed-species biofilms of the Lactobacillus reuteri was determined using MIC assays in 96-well plates. Thereafter, a total of 60 human premolar teeth samples were collected, prepared, and sterilized for ex vivo experiments. All teeth samples were inoculated with mono-species and mixed-species of Enterococcus faecalis and Fusobacterium nucleatum and cultured for 24 hours for biofilm formation. Teeth were divided into 3 groups according to the irrigant type. group 1 was 2.5% NaOCl (positive control), group 2 was saline (negative control) and, group 3 was the experimental reuterin isolate irrigant. The endodontic samples were obtained at 5 minutes and 30 minutes post-irrigation to determine the number of E. faecalis and F. nucleatum. In addition, DNA was extracted and individual bacterial counts were evaluated using Real-time PCR. Data were statistically analyzed via one-way ANOVA, where p<0.05 was set as the level of significance.
Results: Reuterin isolated from L. reuteri showed promising antibiofilm activity. The antibiofilm concentration of reuterin against biofilms of E.faecalis and F.nucleatum biofilms was significant compared to the negative control (p< 0.05) and similar to that of NaOCl positive control. Interestingly, reuterin also demonstrated anti-biofilm activity in ex vivo tooth model at the concentration of 25µg/mL against E. faecalis and F. nucleatum.
Conclusions: The present study has for the first-time demonstrated that reuterin isolate Lactobacillus reuteri can successfully eliminate pathogenic endodontic biofilms as root canal irrigant. This new finding will lead to clinical benefits in future.