IADR Abstract Archives
Reuterin inhibits virulence of Fusobacterium nucleatum ex vivo endodontic biofilms
Objectives: The compound reuterin isolated from the probiotic Lactobacillus reuteri is known to exhibit antimicrobial activity against a wide range of pathogens. The aim of this study was to investigate the effect of reuterin on the expression of fadA, fomA, radD, and aid1 virulence-associated genes in Fusobacterium nucleatum biofilm in an ex vivo root canal model.
Methods: Reuterin was isolated from Lactobacillus reuteri using standard methodology. The minimum inhibitory concentration (MIC) of reuterin against F. nucleatum ATCC-25586 was determined. F. nucleatum cultures were inoculated into sterilized and pre-prepared root canals of a total of 24 human premolar teeth and incubated anaerobically at 37 °C for 24-h. The teeth were divided into four groups based on the type of irrigant used; I. reuterin isolated from L. reuteri LC 382415 (Indonesian strain) 25 ug/mL; II. reuterin isolated from L. reuteri Prodentis (BioGaia); III. NaOCl 2.5 % (positive control); IV. saline (negative control). Root canal models were irrigated with 5 mL of respective irrigating agents for 5-min and 30-min. Thereafter, biofilm samples were collected, subjected to RNA extraction and qPCR using specific-primers for fadA, fomA, radD, and aid1 virulence genes of F. nucleatum. The data were statistically analysed using one-way ANOVA with a significance level of p<0.05.
Results: The MIC of reuterin derived from the Indonesian strain Lactobacillus reuteri against F. nucleatum was found to be 25 μg/mL. At MIC reuterin 25 μg/mL, reuterin significantly inhibited the expression of virulence genes of fadA, fomA, radD, and aid1 genes in the ex vivo F. nucleatum biofilm after 30 min contact time compared to the negative control (p<0.05).
Conclusions: Reuterin exhibits anti-biofilm activity by down-regulating virulence-associated gene expression of E. faecalis ex vivo. Therefore, reuterin may have potential to develop as a endodontic irrigant in future. Further clinical research is required to draw concrete conclusions.
Methods: Reuterin was isolated from Lactobacillus reuteri using standard methodology. The minimum inhibitory concentration (MIC) of reuterin against F. nucleatum ATCC-25586 was determined. F. nucleatum cultures were inoculated into sterilized and pre-prepared root canals of a total of 24 human premolar teeth and incubated anaerobically at 37 °C for 24-h. The teeth were divided into four groups based on the type of irrigant used; I. reuterin isolated from L. reuteri LC 382415 (Indonesian strain) 25 ug/mL; II. reuterin isolated from L. reuteri Prodentis (BioGaia); III. NaOCl 2.5 % (positive control); IV. saline (negative control). Root canal models were irrigated with 5 mL of respective irrigating agents for 5-min and 30-min. Thereafter, biofilm samples were collected, subjected to RNA extraction and qPCR using specific-primers for fadA, fomA, radD, and aid1 virulence genes of F. nucleatum. The data were statistically analysed using one-way ANOVA with a significance level of p<0.05.
Results: The MIC of reuterin derived from the Indonesian strain Lactobacillus reuteri against F. nucleatum was found to be 25 μg/mL. At MIC reuterin 25 μg/mL, reuterin significantly inhibited the expression of virulence genes of fadA, fomA, radD, and aid1 genes in the ex vivo F. nucleatum biofilm after 30 min contact time compared to the negative control (p<0.05).
Conclusions: Reuterin exhibits anti-biofilm activity by down-regulating virulence-associated gene expression of E. faecalis ex vivo. Therefore, reuterin may have potential to develop as a endodontic irrigant in future. Further clinical research is required to draw concrete conclusions.