Inhibition of Dental Biofilm Formation Using Antisense Oligonucleotide to Streptococcus mutans and Streptococcus sobrinus Glucosyltransferases

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Laboratorinė medicina. 2014,
t. 16,
Nr. 4,
p. 165 -
171

Summary

Background. Dental caries remains one of the most prevalent human diseases due to uncontrolled increase of dietary sucrose consumption in modern society. Sucrose is metabolized by Streptococcus mutans and Streptococcus sobrinus to acids causing tooth decay. These streptococci produce glucosyltransferases (Gtfs) for synthesis of sticky glucan polymers from sucrose, which is important for biofilm formation on teeth. In order to reduce dental biofilm build-up and oral diseases it causes, one preventive measure could be blocking of Gtf synthesis.

Aim. This study aim was to test antisense phosphorothioate oligodeo-xyribonucleotide (PS-ODN) targeting simultaneously S. mutans gtfB, gtfC and S. sobrinusgtflmRNAs in order to inhibit biofilm formation in vitro.

Material and methods. Mixed culture of S. mutans and S. sobrinus bacteria were grown anaerobically on glass slides inserted vertically in 24-well cell culture plates containing Todd Hewitt broth with sucrose and sterile saliva under exposure to antisense or missense PS-ODNs at final concentration of 10 ||M. Untreated bacteria served as controls. After 24 h of incubation, glass slides were removed, air-dried and further used for quantitative evaluation of streptococci biofilm applying optical profilometry technique.

Results. It was found that antisense PS-ODN significantly reduced biofilm surface roughness and thickness of mixed S. mutans and S. sobrinus culture suppressing the biofilm development by 1.6-fold overall in comparison to untreated and missense PS-ODN-treated bacteria (p<0.05).

Conclusions. Data demonstrate that antisense PS-ODN considerably attenuate streptococci-induced biofilm build-up on glass slides, and might therefore significantly inhibit dental biofilm formation through simultaneous inactivation of bacterial gtf mRNAs.

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