Identification of a protein-containing enamel matrix layer which bridges with the dentine-enamel junction of adult human teeth

Objective: To investigate the ultrastructure and chemical composition of the dentine-enamel junction and adjacent enamel of minimally processed third molar tooth sections. Design: Undecalcified human third molar erupted teeth were sectioned and etched with 4% EDTA or 37% phosphoric acid prior to visualization by scanning electron microscopy. Confocal Raman spectroscopy was carried out at 50 mu m and more than 400 mu m away from the dentine-enamel junction before and after mild etching. Results: A novel organic protein-containing enamel matrix layer was identified for the first time using scanning electron microscopy of etched bucco-lingual sections of crowns. This layer resembles a three-dimensional fibrous meshwork that is visually distinct from enamel "tufts". Previous studies have generally used harsher solvent conditions which likely removed this layer and precluded its prior characterization. The shape of the organic enamel layer generally reflected that of sheath regions of enamel rods and extended from the dentine-enamel junction about 100-400 mu m into the cuspal enamel. This layer exhibited a Raman C-H stretching peak at similar to 2931 cm(-1) characteristic of proteins and this signal correlated directly with the presence and location of the matrix layer as identified by scanning electron microscopy. Conclusions: The enamel protein layer was most prominent close to the dentine-enamel junction and was largely absent in cuspal enamel >400 mu m away from the dentine enamel junction. We hypothesize that this protein containing matrix layer could provide an important biomechanical linkage between the enamel and the dentine-enamel junction and by extension, with the dentine, of the adult tooth (246 words). (C) 2012 Elsevier Ltd. All rights reserved.