Developing a biological caries model & studying fluoride in caries control

This thesis examines the development of a novel in vitro biological caries model and its suitability in testing the efficacy of anti-caries approaches.Dental caries remains a public health concern worldwide; with extensive treatment costs and impacts on quality of life. Ineffective removal of all dental plaque from tooth surfaces after brushing, the insufficient delivery of anti-caries therapies; along with continuing shifts towards high frequency, sucrose-rich food consumption, expedites the caries disease process. It is, therefore, important to explore caries risk and development at these sites, particularly when representatively assessing the efficacy of a test agent in preventing caries. This caries model enabled the study of the anti-caries effects of fluoride to assess its efficacy in conditions simulating the modern diet. The current methodology employed the Constant Depth Film Fermenter (CDFF) to investigate the caries disease process in response to fluoride delivered continuously; twice and thrice daily; and at different concentrations. The approach is the first in CDFF research modelling caries inclusive of a biologically relevant microcosm biofilm in addition to enamel demineralisation. Specific members of multispecies biofilm were selectively enumerated using traditional microbiological culture techniques whilstcaries was simultaneously quantified with Transverse Microradiography (TMR), Quantitative Light-Induced Fluorescence (QLF), and Non-Contact Surface Profilometry (NCSP). The fluorescence of biofilm illuminated by QLF was also investigated. Results indicated that quantities of total or specific members of the microbial community are not direct indicators of caries risk and turning focus towards the metabolism of oral biofilm bacteria, and how it may be affected, is vital in caries research. TMR and QLF agree when quantifying caries whilst NCSP shows promise in studying surface changes. At 0.05 ppm, fluoride was unable to exert a significant anti-caries effect despite being continuously present during and between sucrose exposures. Laminated lesions confirmed the importance of maintaining elevated levels of fluoride in the oral environment throughout the day. At higher concentrations (1,450 and 228 ppm fluoride) the anti-caries efficacy of fluoride when supplied in frequent applications throughout the day was confirmed. A third application of fluoride did not appear to additively benefit enamel, since all strategies were effective after 10 days regardless of frequency. Nonetheless the increased plaque fluoride reservoir and subtle antimicrobial effects than in twice daily pulsed biofilm, mean the benefit of a third application is likely more discernible in the long term. Scanning Electron Microscopy (SEM) and Energy-Dispersive X-ray Spectroscopy (EDX) elucidated significant calcium fluoride deposits of enamel surfaces beneath biofilm exposed to 1,450 ppm fluoride continuously. In conclusion, the CDFF can produce multispecies biofilm under conditions similar to those of the oral milieu and investigate its cariogenicity in response to diet and experimental anti-caries agents. The model could be examined using an array of techniques to obtain information about aspects of the biofilm, the substratum, and to validate upcoming methods in an orally representative environment. In this regard, the current study contributes not only to enamel caries research but to biofilm research in general by minimising variation.