Chitosan‐based accelerated portland cement promotes dentinogenic/osteogenic differentiation and mineralization activity of shed

Abstract

Calcium silicate‐based cements (CSCs) are widely used in various endodontic treatments to promote wound healing and hard tissue formation. Chitosan‐based accelerated Portland cement (APC‐CT) is a promising and affordable material for endodontic use. This study investigated the effect of APC‐CT on apoptosis, cell attachment, dentinogenic/osteogenic differentiation and mineralization activity of stem cells from human exfoliated deciduous teeth (SHED). APC‐CT was prepared with various concentrations of chitosan (CT) solution (0%, 0.625%, 1.25% and 2.5% (w/v)). Cell attachment was determined by direct contact analysis using field emission scanning electron microscopy (FESEM); while the material extracts were used for the analyses of apoptosis by flow cytometry, dentinogenic/osteogenic marker expression by real‐time PCR and mineralization activity by Alizarin Red and Von Kossa staining. The cells effectively attached to the surfaces of APC and APC‐CT, acquiring flattened elongated and rounded‐shape morphology. Treatment of SHED with APC and APC‐CT extracts showed no apoptotic effect. APC‐CT induced upregulation of DSPP, MEPE, DMP‐1, OPN, OCN, OPG and RANKL expression levels in SHED after 14 days, whereas RUNX2, ALP and COL1A1 expression levels were downregulated. Mineralization assays showed a progressive increase in the formation of calcium deposits in cells with material containing higher CT concentration and with incubation time. In conclusion, APC‐CT is nontoxic and promotes dentinogenic/osteogenic differentiation and mineralization activity of SHED, indicating its regenerative potential as a promising substitute for the commercially available CSCs to induce dentin/bone re-generation.