Formation and assessment of enamel subsurface lesions in vitro

The present study compared two pH-cycling models designed to induce subsurface lesions (SLs) with a less demineralized surface layer on teeth, with the aim of developing new technologies for assessment of such lesions by examining the performance of confocal Raman microscopy for detection of white spot lesions (WSLs). Twelve sound premolars were exposed to two sets of model conditions (A, B) designed to induce SLs. Teeth on which white lesions had formed in vivo were used as positive controls. All specimens were inspected using an intraoral camera and Raman microscopy to detect small changes in the appearance and structure of the enamel. Changes in the natural color of the teeth during the treatment were recorded via the camera. Phosphate maps with their spectra were constructed from the phosphate peak at 960 cm^-1. The depth of lesions was measured on the basis of variations in phosphate peak intensity. Protocol B was reliable for reproducing SLs in a relatively short period. Both protocols had intrinsic limitations in not completely simulating the complex intraoral conditions leading to WSL formation with respect to lesion depth and preservation of an intact surface layer. Raman microscopy can be considered the gold standard for analysis of hard tissue mineralization.