An In Vitro Microbial-Caries Model Used to Study the Efficacy of Antibodies to Streptococcus mutans Surface Proteins in Preventing Dental Caries

Abstract

ABSTRACT The first step for a pathogenic bacterium to initiate infection is via attachment (i.e., through surface determinants) to a suitable receptor. An in vitro microbial artificial-mouth model was used to test the efficacy of polyclonal antibodies to Streptococcus mutans cell surface proteins (CsAb) and a cell surface 59-kDa protein (59Ab) in preventing S. mutans colonization and carious lesion formation. In study 1, groups of 12 human teeth specimens were inoculated with S. mutans , which were incubated with different concentrations of CsAb (A1 [positive control], sterile saline, no antibody; A2, 0.007 mg of antibody protein/ml; and A3, 0.7 mg of antibody protein/ml) for 1 h at 37°C. The negative control group (B1) was not infected and was incubated with Trypticase soy broth (TSB) without dextrose supplemented with 5% sucrose (TSBS). In study 2, the same study design was used except that 59Ab was used instead of CsAb, normal rabbit serum was used in the positive control group (A1), and TSB supplemented with 1% glucose was used as the nutrient to control for sucrose-dependent colonization. All groups were exposed for 4 days to circulating cycles of TSBS and TSB (study 1 and study 2, respectively; 30 min each, three times per day) and a mineral washing solution (21 h per day). Prior to each nutrient cycle, 1 ml of the appropriate CsAb or 59Ab solution was administered to each group and allowed to mix for 30 min before cycling was resumed. Data obtained by confocal laser scanning microscopy demonstrated the presence of a significantly smaller ( P < 0.05) lesion area and a smaller total lesion fluorescence in group A3 than in group A1 for both studies. In study 1, group A2 had significantly smaller values than A1 for lesion depth and area. There were no significant differences between groups A2 and A3 for lesion area or between groups A1 and A2 for total lesion fluorescence. In study 2, there were no significant differences among groups A1 and A2 for lesion depth or between groups A2 and A3 for all of the parameters studied. In both studies, there were no significant differences between S. mutans plaque CFU numbers among any of the groups. These studies demonstrated the efficacy of CsAb and 59Ab in reducing primary caries development in this model, although the underlying mechanism remains unclear.