Dentinal mineralization is not limited in the mineralization front but occurs along with the entire odontoblast process

The mineralization-front theory is historically rooted in mineralization research fields for many decades. This theory is widely used to describe mineralization events in both osteogenesis and dentinogenesis. However, this model does not provide enough evidence to explain how minerals are propagated from the pulp-end dentin to dentin-enamel junction (DEJ). To address this issue, we modified the current research approaches by a) extending the mineral deposition windows of time from minutes to hours, instead of limiting the mineralization assay on days and weeks only; b) switching a regular fluorescent microscope to a more powerful confocal microscope; in which both mineral deposition rates and detail mineral labeling along with dentin tubules can be documented; and c) using reporter mice, including the Gli1-CreERT2 activated tomato and the 2.3 Col1-GFP to mark odontoblast processes combined with mineral dye injections. Our key findings are: 1) Odontoblast-processes, full of numerous mini-branches, evenly spread to entire dentin matrices with a high density of processes and a large diameter of the main process at the predentin-dentin junction; and 2) The minerals deposit along with entire odontoblast-processes and form many individual mineral collars surrounding odontoblast processes. As a result, these merged collars give rise to a single labeled line at the dentin-predentin junction, in which the dental tubules are wider in diameter and denser in odontoblast processes compared to other dentin areas. We therefore propose that it is the odontoblast-process that directly contributes to mineralization, which is not simply limited in the mineralization front at the edge of dentin and predentin, but occurs along with the entire odontoblast process. These new findings will shed new light on our understanding of dentin structure and function, as well as the mechanisms of mineralization.