Dentin formation is largely viewed as the result of interaction of noncollagenous components of the organic matrix with mineral, which leads to the mineral deposition on a collagen scaffold. Although many matrix molecules have been implicated in this process, the necessity of their presence and extent of contribution of each of them to biomineralization remains unknown. The main reason for that is the apparent different rnineralization mechanisms that exist, reflected in dentin by the variation in the composition and properties of organic matrix and the properties of the formed mineral between areas of mantle dentin and circumpulpal dentin. The combination of a high-resolution spectroscopic analytical technique optimized for the analysis of virtually all organic and inorganic mineralization elements with histology provides an excellent means of studying dentin formation. As mantle dentin presents alterations in organic matrix and mineral properties compared to circumpulpal dentin, study of dentin formation in it and comparison to circumpulpal dentin is expected to further help establish the mineralization pathways. For this purpose, the development of a model for study of mantle dentin formation on bovine teeth is suggested. By applying the same methods to dentin from mice that are knockout for a specific matrix macromolecule (at the present time osteopontin, bone sialoprotein, osteonectin, biglycan and decorin) the necessity of each macromolecule for proper dentin formation will be assessed.