Imaging analysis of early DMP1 mediated dentine remineralization

Abstract

ObjectiveThis study assessed the micro-morphological changes in demineralized dentine scaffold following incubation with recombinant dentine matrix protein 1 (rDMP1). DesignExtracted human molar crowns were sectioned into 6 beams (dimensions: 0.50mm×1.70mm×6.00mm), demineralized and incubated overnight in 3 different media (n=4): rDMP1 in bovine serum albumin (BSA), BSA and distilled water. Samples were placed in a chamber with simulated physiological concentrations of calcium and phosphate ions at constant pH 7.4. Samples were immediately processed for transmission electron microscopy (TEM) and field emission-scanning electron microscopy (FE-SEM) after 1 and 2 weeks. ResultsAnalysis of the scaffold showed that decalcification process retained the majority of endogenous proteoglycans and phosphoproteins. rDMP1 treated samples promoted deposition of amorphous calcium phosphate (ACP) precursors and needle shaped hydroxyapatite crystals surrounding collagen fibrils. The BSA group presented ACP bound to collagen with no needle-like apatite crystals. Samples kept in distilled water showed no evidence of ACP and crystal apatite. Results from rDMP1 immobilized on dentine matrix suggests that the acidic protein was able to bind to collagen fibrils and control formation of amorphous calcium phosphate and its subsequent transformation into hydroxyapatite crystals after 2 weeks. ConclusionThese findings suggest a possible bio-inspired strategy to promote remineralization of dentine for reparative and regenerative purposes.