An in vitro study on dentin demineralization and remineralization: Collagen rearrangements and influence on the enucleated phase

Dentin remineralization is of clinical relevance in the therapy of caries and dentin hypersensitivity. This study is aimed at gaining more insights on a molecular scale, through IR spectroscopy, into dentin demineralization and remineralization. The dentin demineralization by ethylenediaminetetraacetic acid, EDTA (17%, 2 h) significantly altered the secondary structure distribution of collagen, upon loss of interaction with calcium ions. To investigate dentin remineralization, previously demineralized human dentin slices were soaked in Dulbecco's Phosphate Buffered Saline (DPBS) or Hank's Balanced Salt Solution HBSS, in close contact with three commercial cements used as sustained releasing sources of Ca²⁺ and OH^- ions (i.e. calcium hydroxide- and calcium silicate-based cements). IR spectroscopy showed the occurrence of remineralization under these conditions. Collagen did not lose its ability to chelate Ca²⁺, and these interactions allowed collagen to rearrange into a conformation similar to that of sound dentin. This process appeared slower in HBSS than DPBS, as also shown by the lower degree of maturation of the inorganic phase enucleated in the former medium (amorphous calcium phosphate versus B-type carbonated apatite). Collagen appeared to act as a spatial constraint to crystal deposition, affecting crystallinity and carbonate content of the enucleated phase. Remineralization was found to strongly depend on the calcium releasing ability of the cements. The fast formation of a rough apatite biocoating may represent a favorable clinical condition in the context of mineralized tissue regeneration.