Secondary caries formation with a two-species biofilm artificial mouth

Abstract

Objectives: The present study aimed to establish Lactobacillus casei in a completely automated Streptococcus mutans-based artificial mouth model and to investigate secondary caries inhibiting properties of glass ionomer cements / resin composite groups in vitro.

Methods: Sixty extracted, caries-free human third molars were used for preparation of standardized class-V-cavities. Specimens were restored with a resin-modified (Photac Fil; PF) as well as a conventional glass ionomer cement (Ketac Molar; KM) and one resin composite bonded with and without conduction of etch-and-rinse technique (Prime&Bond NT, Ceram X mono; C+ERT, C). Following an incubation in distilled water for 28d at 37°C, specimens were exposed to 10,000 thermocycles (+5 °C/+55°C). A completely automated S. mutans-based (DSM No.: 20523) artificial mouth model was extended by establishment of L. casei (DSM No.: 20021). During microbiological loading, demineralization (4h/d) was caused by acid production resulting from bacterial glycolysis and artificial saliva was used for remineralization (20h/d). For quantitative margin analysis under am SEM, epoxy replicas were produced from impressions taken after thermocycling and after microbiological loading. Specimens were cut in half perpendicularly to restoration surfaces and demineralization depths at restoration margins and in 500μm distance from margins were evaluated by means of a fluorescence microscope (FITC filter).

Results: After microbiological loading, overall demineralization depths in enamel at restoration margin (EM) and in 500μm distance (ED) as well as in cementum/dentin at restoration margin (DM) and in 500μm distance (DD) were measured as follows (μm±SD): PF: EM 42±15, ED 60±17, DM 83±18; DD 127±16; KM: EM 46±22, ED 62±17, DM 104±21, DD 143±28; C+ERT: EM 67±19, ED 61±17, DM 165±31, DD 176±35; C: EM 65±23, ED 64±17, DM 161±27, DD 166±33. For the glass ionomer cements, the overall demineralization depths at restoration margins were significantly lower than in 500μm distance from margins (T-test, p<0.05). Especially at restoration margins in cementum/dentin, the resin composite groups showed significantly larger overall demineralizations than the glass ionomer cements (ANOVA, mod. LSD, p<0.05). Decreases of marginal quality were detected in all groups after microbiological loading (Friedman-test, p<0.05).

Significance: The refined experimental setup was suitable for production of artificial secondary caries-like lesions. Glass ionomer cements as fluoride-releasing materials may show an inhibition of secondary caries formation to a certain extent.

Keywords: Automated artificial mouth model; Biofilm; Glass ionomer cement; L. casei; Resin-modified glass ionomer cement; S. mutans; Secondary caries.