Synchrotron Radiation (SR) Wide-angle X-ray Diffraction (WAXD) and Small-angle X-ray Scattering (SAXS) techniques were used to assess microstructure of bovine enamel white-spot lesions (WSL) evaluated in a 10-day pH cycling model comprising three different dentifrice groups: (A) 0.21% NaF plus TCP (Clinpro Tooth Crème), (B) 1.1% NaF plus TCP (Clinpro 5000), or (C) 0% NaF (Tom’s of Maine) dentifrice. Each day consisted of four 2-minute treatments, one 4-hour acid challenge (pH=5.0), and immersion in artificial saliva (pH=7.0) between these events. These specimens were also examined with cross-sectional microhardness, digital light microscopy and FE-SEM (field emission scanning electron microscope), and demonstrated the remineralization model effected changes in subsurface microstructure. X-ray diffraction data from WAXD and SAXS were collected on enamel slab cross-sections extending from 0 μm to 150 μm, in 6 μm microbeam increments. A primary outcome of this observational study was that simultaneous WAXD and SAXS measurements were able to resolve significant differences (ANOVA, Student’s t-test, p<0.05) between the effects of the two fluoridecontaining dentifrices on subsurface lesion microstructure. In particular, enamel lesions treated with 1.1% NaF dentifrice group manifested an abundance of nanometer-sized crystallites, while treatment with the 0.21% NaF dentifrice produced larger apatite-like crystals. While the presence of fluoride in both cases promoted regularity in crystal size and orientation, this was not observed for lesions treated without fluoride. Altogether, our observations demonstrate the pathological processes for remineralization are markedly influenced by the presence and concentration of fluoride, the microstructural characteristics of which can be distinguished using the simultaneous WAXD and SAXS technique.

Synchrotron radiation micro-computed tomography (SR micro-CT), considered superior to standard polychromatic micro-CT techniques, was used to assess the densities of bovine enamel white-spot lesions (WSL) treated in a 10-day pH cycling model with either: (A) Clinpro Tooth Crème (0.21% NaF plus TCP), (B) Clinpro 5000 (1.1% NaF plus TCP) or (C) Tom's of Maine (0% NaF) dentifrice. Each day consisted of four 2 min treatments, one 4 h acid challenge (pH=5.0), and immersion in artificial saliva (pH=7.0) between these events. After 10 days, WSL specimens were evaluated for lesion depth using confocal microscopy and lesion density using SR micro-CT with depths ranging from 2.76 to 113.16 μm, in 2.76 μm slice increments. Statistical analyses (Student's t-test) were performed at the 95% confidence level. SR micro-CT analyses revealed the NaF plus TCP dentifrices improved WSL densities relative to the fluoride-free toothpaste, and is consistent with an earlier study utilizing polychromatic micro-CT. In contrast to previous findings, SR micro-CT analyses also revealed significant differences in WSL densities treated with the two NaF dentifrices at enamel depths of 13.80, 16.56, and 19.32 μm. These findings suggest SR micro-CT may be especially suited for detecting density differences in lesions sensitive to fluoride-driven remineralization processes.