Antimicrobial Effect of Dental Adhesive on Cariogenic Multi-Species Biofilm

Abstract

Dental caries, or tooth decay, is one of the most widespread chronic and multifactorial diseases affecting people worldwide. It is described as the localized destruction of tooth structure by acids produced from bacteria fermentation of edible carbohydrates. The disease process begins within the bacterial biofilm on the surfaces of the teeth. Streptococcus sanguinis, Sreptococcusgordonii, and Streptococcus mutans are the main organisms in the biofilm associated with health or disease conditions in the oral cavity. Streptococcus mutans is believed to be the primary cause of enamel demineralization and the development of dental caries. S. gordonii and S. sanguinis belong to a group of streptococci considered health-related commensal oral bacteria, crucial for forming a biofilm on oral hard tissues. Several types of materials with multipurpose characteristics have been incorporated into dental adhesives. Recently, the calcium salt of an acidic monomer (calcium salt of 4-methacryloxyethyl trimellitic acid, or CMET) has been included in dental adhesive and demonstrated to suppress cariogenic biofilm formation of S. mutans. However, using a single-species model may not mimic the intricate antagonistic and synergistic relations that occur in oral biofilms. Therefore, the effect on cariogenic multi-species biofilm was assessed in this study. Dental adhesive consisting of CMET and 10-methacryloyloxydecyl dihydrogen calcium phosphate (MDCP) (Bio-Coat CA) was spread over the flat-bottom surface of the 96-well plate and LED light-cured. Then it was coated with sterile saliva at 37 °C for 45 min to form an acquired pellicle for microbial attachment. The multi-species bacterial suspension containing Streptococcus mutans ATCC 25715, Streptococcus sanguinis ATCC 10556, and Streptococcus gordonii ATCC 10558 was prepared and added to the saliva-coated well. Then, the plate was incubated at 37°C in a 5% CO2 atmosphere for 24 h, 48 h, and 72 h to support the biofilm formation. The number of vital bacteria in the biofilm was determined with the WST-8 Microbial Cell Counting Kit (Dojindo Molecular Technologies, USA). All tests were done in triplicate and repeated three times. For statistical analysis, Kruskal-Wallis and Dunn’s tests were employed. The results showed that at 24 h, 48 h, and 72 h, dental adhesive with CMET could inhibit the biofilm formation of multi-species bacteria significantly compared with controls. The percentages of biofilm inhibition were 29.1%, 34.7%, and 33.2% at 24 h, 48 h, and 72 h, respectively. Dental adhesive containing CMET displayed favorable multi-species biofilm-inhibiting effects up to 72 h of biofilm growth. It is a promising adhesive for use to prevent secondary caries at the sites of restorations.