Candida species in intact in vivo biofilm from carious lesions

Abstract

Objectives: The aim of this study was to investigate the occurrence and spatial localization of Candida species in intact in vivo biofilm from caries lesions in root and occlusal surfaces.

Materials and methods: Biofilm from 7 teeth with root caries and 9 teeth with occlusal caries were analyzed. The teeth were fixed, embedded, sectioned and decalcified before fluorescence in situ hybridization using oligonucleotide probes against all bacteria (EUB338), all yeast (PF2), streptococci (STR405, MUT590) and C. albicans (Du.al 1249). Sections were analyzed using fluorescence microscopy.

Results: Yeasts, most of which were C. albicans or C. dubliniensis, colonized occlusal and root surface sites and exhibited both yeast and hyphal forms. Two characteristic colonization patterns were apparent: In one pattern, the biofilm presented defined areas comprising Candida hyphal networks mixed with filamentous, rod-like and coccoid bacteria, often extending through the entire biofilm thickness. In the other pattern, Candida formed corncob configurations with non-mutans streptococci in the biofilm surface layer. Mutans streptococci formed isolated colonies and did not typically appear in close proximity with Candida.

Conclusion: For the first time, we showed that C. albicans and C. dubliniensis are integrated components of caries biofilm architecture. Co-localization of Candida and non-mutans streptococci indicates a beneficial interaction between these organisms. Diverse metabolic properties of Candida spp. suggest that these organisms may influence the ecology of cross-kingdom microbial communities in caries.

Keywords: Biofilms; Candida. Fungi; Dental caries; Microbiology; Structural composition.