Chlorhexidine has long been used in mouth washes for the control of dental caries, gingivitis and dental plaque. Minimal inhibitory concentration (MIC) is the lowest concentration of an antimicrobial substance to inhibit the growth of bacteria. Concentrations lower than the MIC are called sub minimal inhibitory concentrations (sub-MICs). Many studies have reported that sub-MICs of antimicrobial substances can affect the virulence of bacteria. The aim of this study was to investigate the effect of sub-MIC chlorhexidine on biofilm formation and coaggregation of oral early colonizers, such as Streptococcus gordonii, Actinomyces naeslundii and Actinomyces odontolyticus. The biofilm formation of S. gordonii, A. naeslundii and A. odontolyticus was not affected by sub-MIC chlorhexidine. However, the biofilm formation of S. mutans increased after incubation with sub-MIC chlorhexidine. In addition, cell surface hydrophobicity of S. mutans treated with sub-MIC of chlorhexidine, decreased when compared with the group not treated with chlorhexidine. However, significant differences were seen with other bacteria. Coaggregation of A. naeslundii with A. odontolyticus reduced by sub-MIC chlorhexidine, whereas the coaggreagation of A. naeslundii with S. gordonii remained unaffected. These results indicate that sub-MIC chlorhexidine could influence the binding properties, such as biofilm formation, hydrophobicity and coaggregation, in early colonizing streptococci and actinomycetes.

Minimal inhibitory concentration (MIC) is the lowest antibiotic concentration that inhibits the visible growth of bacteria. Sub-minimal inhibitory concentration (Sub-MIC) is defined as the concentration of an antimicrobial agent that does not have an effect on bacterial growth but can alter bacterial biochemistry, thus reducing bacterial virulence. Many studies have confirmed that sub-MICs of antibiotics can inhibit bacterial virulence factors. However, most studies were focused on Gram-negative bacteria, while few studies on the effect of sub-MICs of antibiotics on Gram-positive bacteria. In this study, we examined the influence of sub-MICs of doxycycline, tetracycline, penicillin and amoxicillin on biofilm formation and coaggregation of Streptococcus gordonii, Streptococcus mutans, Actinomyces naeslundii, and Actinomyces odontolyticus. In this study, incubation with sub-MIC of antibiotics had no effect on the biofilm formation of S. gordonii and A. naeslundii. However, S. mutans showed increased biofilm formation after incubation with sub-MIC amoxicillin and penicillin. Also, the biofilm formation of A. odontolyticus was increased after incubating with sub-MIC penicillin. Coaggregation of A. naeslundii with S. gordonii and A. odontolyticus was diminished by sub-MIC amoxicillin. These observations indicated that sub-MICs of antibiotics could affect variable virulence properties such as biofilm formation and coaggregation in Gram-positive oral bacteria.