Advanced imaging of dentin microstructure

Abstract

This study aimed to demonstrate the feasibility of using optical coherence tomography (OCT) for locating the sectioning site of a specimen before characterizing the ultrastructural features of dentin surfaces as well as the inner wall of the dentinal tubules (DT) using a field emission scanning electron microscope (FESEM). Eight sound human molar teeth were extracted, examined via cross-polarization optical coherence tomography (CP-OCT), embedded, and hemisectioned using a low-speed diamond sawing machine. Next, each sectioned surface was further trimmed, polished, and examined under a confocal laser scanning microscope (CLSM) to locate the target area on the superficial dentin. Subsequently, each section was gold-coated and examined using FESEM. Backscattered reflection from the dentin layer was less than that from the enamel under CP-OCT. Distinct reflections from certain enamel and dentin microstructures were observed before sectioning the specimens. Areas with enamel cracks and dentin defects were identified and avoided during sectioning. At the micron level, the CLSM images exhibited a homogenous distribution of the DT orifices. Low magnification FESEM images showed intertubular dentin as a loosely condensed globular layer with shallow grooves in between, whereas peritubular dentin exhibited more organized condensation of apatite crystals surrounding the DT orifices. High magnification of the DT revealed a cross-linking layer of mineralized collagen network extending in the peri-intratubular lumen, with scattered globules of matrix vesicles. CP-OCT enabled the realization of rapid initial scanning and image acquisition with high contrast at the micron scale before profound insights into dentin ultrastructures at the nano scale were provided by FESEM. The variations in structural densities of the dental tissues significantly affected the image contrast and helped identify underlying structures.