A Novel in vitro Microbial-Based Model for Studying Caries Formation – Development and Initial Testing

We report on the development and the initial testing of a new microbial-based caries model. Specimens were fixed on a rotating mount within a reaction chamber hermetically surrounded by a sterilised glove box. A cariogenic environment was obtained by inoculation with Streptococcus mutans (ATCC 25175) combined with a continuously repeating supply of sucrose solution, trypticase soy broth and artificial saliva applied by dripping. Twenty-five caries-free upper premolars were used. The mesial parts of the occlusal fissures had been sealed with a resin-based fissure sealant (test group 1). To produce marginal gaps, the distal parts had been moistened with saliva before resin application (test group 2). Five teeth served as control and were exposed to all fluids under sterile conditions before being removed from the system after 7 days. Test specimens were infected with S. mutans and were incubated for another 14 days. No unintentional contamination occurred during the 3-week period of operation. Demineralizations were evaluated by using confocal laser scanning microscopy. Only the test specimens showed clearly visible signs of biofilm formation and caries-like lesions. The mean primary lesion depth did not differ significantly between test groups. Wall lesion depths and surface areas of demineralizations underneath the fissure sealants were significantly higher in test group 2. Thus, our model allows the simultaneous production of primary and secondary caries-like enamel lesions in a considerable number of specimens and facilitates the possibility to manipulate and transfer them without necessarily terminating the experiment, opening new possibilities for in vitro caries research.