Direct or in situ methods (e.g., confocal microscopy, microsensors) are used to study non-oral biofilms for almost two decades, and they have recently been introduced in the research of dental plaque. We combined a pH microsensor technique and transversal microradiography (TMR) in a pilot study where the effects of nutrient availability on plaque acidogenicity and on the development of caries-like lesions were assessed. One volunteer accumulated dental plaque for 7 days in 0.2-mm-wide and 0.8-mm-deep dentin grooves at four conditions: (1) saliva only, (2) 8 × 5 min/day dipping in 10% sucrose solution, (3) 7 × 5 min/day consumption of sweet cookies plus one meal/day, and (4) subject’s regular diet. Plaque pH versus time and depth profiles in the grooves were recorded ex vivo before and after sugar challenge. ‘Saliva’ plaque responded to sugar with slow pH decrease – minimum pH 5.6–5.8 was reached after 30 min, while sugar dipping resulted in metabolically active plaque (minimum pH 5.3–5.5 within 4–8 min). TMR analysis revealed no demineralization after these two periods. Metabolically active plaque leading to distinct lesions resulted from frequent plaque exposure to diets rich in starch and carbohydrates (groups 3 and 4). These findings strengthen the view that the plaque acidogenicity does not necessarily reflect cariogenicity, and that retention of food components may account for increased cariogenicity at plaque retention sites in the mouth.

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