A novel non-destructive technique for qualitative and quantitative measurement of dental erosion in its entirety by porosity and bulk tissue-loss.

OBJECTIVE: To explore the potential of combining non-contact profilometry (NCP) and confocal laser scanning microscopy (CLSM) data to measure the entire erosive process non-destructively and to validate findings using inductively coupled plasma-atomic emission spectroscopy (ICP-AES), scanning electron microscopy (SEM) and surface microhardness (SMH) using the same samples throughout. METHODS: Polished bovine enamel samples (n = 35) were divided into groups (7/group) with similar SMH values. Samples underwent individual erosive challenges (1 % citric acid, pH3.8) for 1, 5, 10, 15 or 30 min under stirring and aliquot extracts were analysed for Ca and P by ICP-AES. SMH was used to measure erosive softening. Profilometry was used to assess bulk volume loss (BVL). Images were captured by SEM. Samples were stained with rhodamine-B (0.1 mM, 24 h) and images captured by CLSM. Image processing was used to determine changes in fluorescent volume for the first 10 µm (ΔFV10) for each enamel sample which were combined with BVL to calculate total lesion volume (TLV). ANOVA, linear regression and Pearson correlation analysis were used where applicable. RESULTS: Surface softening, [Ca], [P], BVL and ΔFV10µm increased with acid erosion duration which were significant by 10 min (P < .01). The Ca:P ratio increased to 1.57 then decreased after 5 min erosion suggesting a sub/surface phase change, which was observed by SEM and CLSM showing significant changes to the enamel surface and subsurface morphology with time. Combination of BVL and ΔFV10 as TLV strengthened the significant correlations with [Ca], [P], and SMH (P < .01). CONCLUSION: This novel combination of CLSM and NCP allows for concurrent non-destructive quantification of the entire erosive process by mineral loss, and qualitatively characterise microstructural changes during dental erosion.

Characterisation of mineral loss as a function of depth using confocal laser scanning microscopy to study erosive lesions in enamel: A novel non-destructive image processing model.

OBJECTIVE: The aim was to develop a novel image processing protocol for confocal laser scanning microscopy (CLSM) to study mineral distribution within erosive lesions as a function of depth. METHODS: Polished bovine enamel samples (n = 80) were divided into groups (8/group) with similar mean surface microhardness (SMH) values. Samples underwent erosion (1 % citric acid pH3.8) for 1,5,10,15, or 30 min, with or without stirring giving 10 treatment groups in a 2*5 factorial design. SMH was used to measure erosive softening. Profilometry was used to measure bulk tissue loss. Samples were then stained with rhodamine-B (0.1 mM, 24 h) and imaged using CLSM. Image processing was used to measure fluorescence volume (FV) as a function of depth for each image. The data from reference images were subtracted from post-erosive data to determine changes in fluorescent volume (ΔFV) as a function of depth. 2-way ANOVA and linear regression analysis were used where applicable. RESULTS: Surface softening and bulk tissue loss increased with acid erosion duration with or without stirring. Stirring significantly increased net softening at each time point; specimens underwent significantly more bulk tissue loss (P < 0.05). CLSM showed the erosive lesion deepened as exposure to acid increased, and that at the near surface (0-10 µm) FV and ΔFV increased rapidly for stirred solutions. The increase in pore space translated to a softer surface as measured by SMH. CONCLUSION: This novel non-destructive method allows concurrent quantification of dental erosion by mineral loss as a function of depth, and qualitative characterisation of microstructural changes during early erosion.