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Κυριακή 4 Δεκεμβρίου 2022

Tideglusib enhances odontogenic differentiation in human dental pulp stem cells in vitro

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Abstract

Aim

Tideglusib is a small molecule agonist of the canonical Wnt pathway. The present study investigated the influence of Tideglusib on human dental pulp stem cell (hDPSC) proliferation, apoptosis, migration, and odonto/osteogenic differentiation.

Methodology

hDPSCs were treated with 50 nM, 100 nM, or 200 nM Tideglusib. β-catenin accumulation was detected by immunofluorescence staining. Colony forming unit ability was assessed by staining with Coomassie blue. Cell cycle progression and cell apoptosis were investigated using flow cytometry. Cell migration was examined using an in vitro wound healing assay. Osteogenic differentiation was examined using alkaline phosphatase (ALP) staining, alizarin red s staining, and osteogenic-related gene expression. The gene expression profile was examined using a high throughput RNA sequencing technique. All experiments were repeated using cells derived from at least four different donors (n = 4). The Mann Whitney U test was used to identify significant differences for two independent group comparisons. For three or more group comparison, statistical differences were assessed using the Kruskal Wallis test followed by a pairwise comparison. The significance level was set at 5% (p< /i> < 0.05).

Results

Tideglusib activated the Wnt signaling pathway in hDPSCs as demonstrated by an increase in cytoplasmic β-catenin accumulation and nuclear translocation. Tideglusib did not affect hDPSC proliferation, cell cycle progression, cell apoptosis, or cell migration. In contrast, 50 nM and 100 nM Tideglusib significantly enhanced mineralisation and osteogenic marker gene expression (RUNX2, ALP, BMP2, and DSPP) (p < 0.05).

Conclusions

Tideglusib enhanced the odonto/osteogenic differentiation of hDPSCs. Therefore, incorporating this bioactive molecule in a pulp capping material could be a promising strategy to promote dentine repair.

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