IADR Abstract Archives
Reuterin of Lactobacillus-reuteri Indonesian Strain Reduces Oral Pathogens Biofilm Formation
Objectives: To evaluate the potentiate of reuterin isolate Lactobacillus reuteri Indonesian strain in inhibiting monospecies and multispecies biofilm Streptococcus mutans and Porphyromonas gingivalis
Methods: L. reuteri was isolated from saliva of 20 Indonesians, and the species was confirmed by PCR, using 16S rRNA gene primers. To produce reuterin, the isolate was mixed in glycerol-containing MRS broth, and grown in anaerobic condition for 3h. The molecule’s weight of reuterin was counted by SDS-PAGE. Reuterin effect on biofilm formation of monospesies or multispecies (S. mutans ATCC 15175 and P. gingivalis ATCC 33277) was evaluated by biofilm assay in vitro. Furthermore, 200µL of S. mutans and P. gingivalis (1x107 cfu/mL) was added into microtitre plate and incubated for 48h to form biofilm. Reuterin concentrations 100%, 50%, 25%,and 12.5% was added into each well, and its inhibitor effect was observed after 1h, 3h, 6h, and 24h. Chlorhexidine (1,2%) was used as positive control. The biofilm mass was calculated by crystal violet stain (0.5% w/v) by measuring the Optical Density at 490 nm and 655 nm, respectively.
Results: PCR assays showed that the clinical isolates were L. reuteri, while 52 kDa band on SDS-PAGE confirmed the presence of reuterin. Data showed that reuterin had the ability to inhibit biofilm formation, either as monospecies or multispecies. We found that reuterin in all concentrations and all time periods set in this study had the ability to reduce biofilm of both species (p < 0.05). However, the most effective concentration in inhibiting biofilm was 25%. Additionally, the best incubation time in reducing biofilm was 3 hours period.
Conclusions: This study showed the antibiofilm activity of reuterin produced by L.reuteri isolated from Indonesian subject have the capacity to reduce biofilm formation of two species related to caries and periodontal diseases bacteria. More studies are needed to explore this effect.
Methods: L. reuteri was isolated from saliva of 20 Indonesians, and the species was confirmed by PCR, using 16S rRNA gene primers. To produce reuterin, the isolate was mixed in glycerol-containing MRS broth, and grown in anaerobic condition for 3h. The molecule’s weight of reuterin was counted by SDS-PAGE. Reuterin effect on biofilm formation of monospesies or multispecies (S. mutans ATCC 15175 and P. gingivalis ATCC 33277) was evaluated by biofilm assay in vitro. Furthermore, 200µL of S. mutans and P. gingivalis (1x107 cfu/mL) was added into microtitre plate and incubated for 48h to form biofilm. Reuterin concentrations 100%, 50%, 25%,and 12.5% was added into each well, and its inhibitor effect was observed after 1h, 3h, 6h, and 24h. Chlorhexidine (1,2%) was used as positive control. The biofilm mass was calculated by crystal violet stain (0.5% w/v) by measuring the Optical Density at 490 nm and 655 nm, respectively.
Results: PCR assays showed that the clinical isolates were L. reuteri, while 52 kDa band on SDS-PAGE confirmed the presence of reuterin. Data showed that reuterin had the ability to inhibit biofilm formation, either as monospecies or multispecies. We found that reuterin in all concentrations and all time periods set in this study had the ability to reduce biofilm of both species (p < 0.05). However, the most effective concentration in inhibiting biofilm was 25%. Additionally, the best incubation time in reducing biofilm was 3 hours period.
Conclusions: This study showed the antibiofilm activity of reuterin produced by L.reuteri isolated from Indonesian subject have the capacity to reduce biofilm formation of two species related to caries and periodontal diseases bacteria. More studies are needed to explore this effect.