IADR Abstract Archives
Indonesian Strain Lactobacillus reuteri-derived Reuterin Reduces Interleukin-8 Expression of Inflamed-HaCat Cells
Objectives: This study was conducted to investigate the potential of Indonesian strain Lactobacillus reuteri-derived reuterin in reducing interleukin (IL)-8 mRNA expression of Streptococcus mutans- or Porphyromonas gingivalis-inflamed HaCat cells.
Methods: L. reuteri was isolated from saliva of 20 Indonesian subjects, and confirmed by primering 16S rRNA genes with PCR. To produce reuterin, isolated bacteria were cultured with glycerol-supplemented MRS broth in anaerobic condition for 3 hours. To inflame HaCat cells, 107 CFU/mL S. mutans (ATCC-25175) or P. gingivalis (ATCC-33277) were preheated at 80°C for 30 minutes and inoculated into 105 cells/mL HaCat cells culture. After that, HaCat cells were treated with 12.5, 25, 50 or 100% reuterin for 1, 3, 6 or 24 hours at 37°C with 5% CO2 . Total HaCat cells RNAs were extracted and reverse transcripted. Resulted cDNAs were then analyzed and quantified for IL-8 mRNA expression with qPCR. Additionally, 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide (MTT) assay was applied in order to evaluate the cytotoxic effect of reuterin on HaCat cells. Statitical analysis was conducted with ANOVA test and significant level was set at p<0.05.
Results: L. reuteri was successfully isolated from 6 subjects and confirmed. IL-8 mRNA expression was increased in S. mutans or P. gingivalis-inflamed HaCat cells. Treatment of reuterin in all concentrations and time periods could reduce IL-8 mRNA expression significantly (p<0.05). The most effective concentration in reducing IL-8 mRNA expression was 12.5% and the best incubation period was 24 hours.. Lowest cell viability in this research is 93%, which suggests that reuterin isolated from L. reuteri Indonesian strain does not cause toxicity to epithelial cells.
Conclusions: Indonesian strain Lactobacillus reuteri-derived reuterin reduced interleukin (IL)-8 mRNA expression of Streptococcus mutans- or Porphyromonas gingivalis-inflamed HaCat cells. Further studies are still needed to explore this effect.
Methods: L. reuteri was isolated from saliva of 20 Indonesian subjects, and confirmed by primering 16S rRNA genes with PCR. To produce reuterin, isolated bacteria were cultured with glycerol-supplemented MRS broth in anaerobic condition for 3 hours. To inflame HaCat cells, 107 CFU/mL S. mutans (ATCC-25175) or P. gingivalis (ATCC-33277) were preheated at 80°C for 30 minutes and inoculated into 105 cells/mL HaCat cells culture. After that, HaCat cells were treated with 12.5, 25, 50 or 100% reuterin for 1, 3, 6 or 24 hours at 37°C with 5% CO2 . Total HaCat cells RNAs were extracted and reverse transcripted. Resulted cDNAs were then analyzed and quantified for IL-8 mRNA expression with qPCR. Additionally, 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide (MTT) assay was applied in order to evaluate the cytotoxic effect of reuterin on HaCat cells. Statitical analysis was conducted with ANOVA test and significant level was set at p<0.05.
Results: L. reuteri was successfully isolated from 6 subjects and confirmed. IL-8 mRNA expression was increased in S. mutans or P. gingivalis-inflamed HaCat cells. Treatment of reuterin in all concentrations and time periods could reduce IL-8 mRNA expression significantly (p<0.05). The most effective concentration in reducing IL-8 mRNA expression was 12.5% and the best incubation period was 24 hours.. Lowest cell viability in this research is 93%, which suggests that reuterin isolated from L. reuteri Indonesian strain does not cause toxicity to epithelial cells.
Conclusions: Indonesian strain Lactobacillus reuteri-derived reuterin reduced interleukin (IL)-8 mRNA expression of Streptococcus mutans- or Porphyromonas gingivalis-inflamed HaCat cells. Further studies are still needed to explore this effect.