IADR Abstract Archives
Antisense Oligonucleotide in Fluoride Mouthwash Inhibits Mutans Streptococci Biofilm Formation
Objectives: To test if a new biofilm-inhibiting antisense phosphorothioate oligodeoxyribonucleotide (PS-ODN) targeting simultaneously the selected homologous region of Streptococcus mutans glucosyltransferase (gtf) B, gtfC and Streptococcus sobrinus gtfI mRNAs is compatible with a sodium fluoride (NaF) mouthwash in suppressing streptococci biofilm formation in vitro.
Methods: Mixed cultures of S. mutans UA159 and S. sobrinus SL1 bacteria were grown anaerobically on vertical glass slides in 24-well cell culture plates containing Todd Hewitt broth with 1% sucrose and sterile-filtered human saliva (1:10) or 10% heat-inactivated horse serum under exposure to 10 µM of PS-ODN (5′-GCAGACCATTGCTTAATCT-3′) and 22.6 ppm of fluoride resulting from mouthwash (% w/v): 0.06% PS-ODN, 0.05% NaF, 0.1% saccharin-Na, 0.2% menthol. Untreated bacteria and bacteria exposed to mouthwash without PS-ODN, served as controls. After 24 h of incubation, glass slides were air-dried and further used for quantitative evaluation of biofilm applying optical profiler Sensofar PLµ 2300 and Gwyddion 2.27 software. Data obtained from two experiments (n=12, surface roughness; n=10, biofilm thickness) were analyzed using SPSS 23.0 program.
Results: The PS-ODN in combination with NaF mouthwash decreased surface roughness (parameter Rq) of the biofilm-covered glass slides by 62% and 75% versus untreated bacteria (p<0.05) in medium containing saliva and serum, respectively. This combination also reduced biofilm thickness by 88% and 78% versus untreated bacteria (p<0.05) in medium with saliva and serum, respectively. Mouthwash alone decreased roughness (parameter Rq) by 23% compared to untreated bacteria (p<0.05) growing in presence of saliva but not in serum-containing medium (p>0.05), whereas it reduced biofilm thickness by 38% and 22% versus untreated bacteria (p<0.05) in medium with saliva and serum, respectively.
Conclusions: The antisense PS-ODN is compatible with a NaF mouthwash and significantly inhibits mutans group streptococci biofilm formation on glass, and the combination might therefore be used as antibiofilm and/or anticaries agent in mouthwash formulations.
Methods: Mixed cultures of S. mutans UA159 and S. sobrinus SL1 bacteria were grown anaerobically on vertical glass slides in 24-well cell culture plates containing Todd Hewitt broth with 1% sucrose and sterile-filtered human saliva (1:10) or 10% heat-inactivated horse serum under exposure to 10 µM of PS-ODN (5′-GCAGACCATTGCTTAATCT-3′) and 22.6 ppm of fluoride resulting from mouthwash (% w/v): 0.06% PS-ODN, 0.05% NaF, 0.1% saccharin-Na, 0.2% menthol. Untreated bacteria and bacteria exposed to mouthwash without PS-ODN, served as controls. After 24 h of incubation, glass slides were air-dried and further used for quantitative evaluation of biofilm applying optical profiler Sensofar PLµ 2300 and Gwyddion 2.27 software. Data obtained from two experiments (n=12, surface roughness; n=10, biofilm thickness) were analyzed using SPSS 23.0 program.
Results: The PS-ODN in combination with NaF mouthwash decreased surface roughness (parameter Rq) of the biofilm-covered glass slides by 62% and 75% versus untreated bacteria (p<0.05) in medium containing saliva and serum, respectively. This combination also reduced biofilm thickness by 88% and 78% versus untreated bacteria (p<0.05) in medium with saliva and serum, respectively. Mouthwash alone decreased roughness (parameter Rq) by 23% compared to untreated bacteria (p<0.05) growing in presence of saliva but not in serum-containing medium (p>0.05), whereas it reduced biofilm thickness by 38% and 22% versus untreated bacteria (p<0.05) in medium with saliva and serum, respectively.
Conclusions: The antisense PS-ODN is compatible with a NaF mouthwash and significantly inhibits mutans group streptococci biofilm formation on glass, and the combination might therefore be used as antibiofilm and/or anticaries agent in mouthwash formulations.