Abstract
BackgroundResearch shows that nano-bioceramics can modulate the differentiation of dental stem cells. The novel ready-to-use calcium-silicate-based root-canal sealer iRoot SP is widely used in root filling. Accordingly, the aim of this study was to evaluate the effects of iRoot SP on proliferation and osteogenic differentiation in human stem cells from the apical papilla (hSCAPs).MethodshSCAPs were isolated and characterized in vitro, then cultured with various concentrations of iRoot SP extract. Cell proliferation was assessed by CCK-8 assay, and scratch-wound-healing assays were performed to evaluate cell-migration capacity. hSCAPs were then cultured in osteogenic medium supplemented with iRoot SP extracts. Alkaline phosphatase (ALP) activity assay was used to evaluate ALP enzyme levels. Alizarin red staining and cetylpyridinium chloride (CPC) assays were performed to assess calcified-nodule formation and matrix-calcium accumulation of hSCAPs. The mRNA and protein expression levels of the osteogenic markers OCN, OSX, Runx2, and DSPP were determined by qRT-PCR and Western blotting. The data were analyzed using one-way ANOVA and LSD-t tests.ResultsiRoot SP at low concentrations (2, 0.2, and 0.02 mg/mL) is nontoxic to hSCAPs. iRoot SP at concentrations of 0.02 and 0.2 mg/mL significantly increases cell-migration capacity. In terms of osteogenic differentiation, 0.2 mg/mL iRoot SP promotes intracellular ALP activity and the formation of mineralized nodules. Moreover, the expression of osteogenic markers at the mRNA and protein levels are upregulated by iRoot SP.ConclusioniRoot SP is an effective filling material for periapical bone regeneration.
Highlights
Research shows that nano-bioceramics can modulate the differentiation of dental stem cells
Alizarin red staining and cetylpyridinium chloride (CPC) assays demonstrated that hSCAPs exposed to iRoot SP extract at 0.2 mg/mL generate more mineralized nodules and present higher calcium contents compared with the control group after 14 and 21 days (P < 0.01) (Fig. 3B, C), but no significant effects are observed at 3 and 7 days
The results showed that 0.2 mg/mL iRoot SP extract significantly increases the Alkaline phosphatase (ALP) activity of hSCAPs, and Alizarin red staining and semi-quantitative CPC analysis demonstrated that the calcium matrix produced by cells increases significantly under treatment at this optimal concentration
Summary
Research shows that nano-bioceramics can modulate the differentiation of dental stem cells. The aim of this study was to evaluate the effects of iRoot SP on proliferation and osteogenic differentiation in human stem cells from the apical papilla (hSCAPs). Pulpal and periapical diseases are common endodontic conditions, and root canal therapy (RCT) is currently the most effective treatment for such conditions [1]. Human stem cells from the apical papilla (hSCAPs) are a type of dental mesenchymal stem cells first reported by Sonoyama et al [6]. They exist in the undeveloped apical area of young permanent teeth and exhibit superior. HSCAPs show great promise for use in the regeneration of dentin-bone-like tissues after pulp and periapical disease treatment as well as having potential clinical value as seed cells in bone-tissue regeneration [10, 11]
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