Wnt/beta-catenin plays a dual function in calcium hydroxide induced proliferation, migration, osteogenic differentiation and mineralization in vitro human dental pulp stem cells

Aim: Calcium hydroxide is the gold standard material for pulp capping and has been widely used in clinical dentistry. Calcium hydroxide promotes proliferation, migration and osteogenic differentiation of dental pulp stem cells (DPSCs). However, the underlying mechanism is not clear. Our study investigated the role of Wnt/beta-catenin pathway in calcium hydroxide-induced proliferation, migration, osteogenic differentiation and mineralization of human DPSCs. Methodology: Protein and gene expression was detected by western blot (WB), immunofluorescence staining and quantitative real-time PCR (qPCR). Cell viability was analysed using the Cell Counting Kit-8 (CCK-8) assay. Wound-healing assay was used to analyse cell migration. The expression of alkaline phosphatase (ALP) was detected using ALP staining. Mineralization was analysed by alizarin red staining. Results: Calcium hydroxide increased the protein expression of phosphorylated-GSK3 beta/GSK3 beta, beta-catenin and the gene expression of LEF-1. Inhibition of Wnt/beta-catenin abolished calcium hydroxide-induced proliferation and migration of DPSCs in 24 h. However, incubation with calcium hydroxide for 7 days and 14 days reduced Wnt/beta-catenin signalling. Inhibition of Wnt/beta-catenin promoted calcium hydroxide-induced osteogenic differentiation and mineralization in DPSCs. Conclusion: Wnt/beta-catenin pathway plays a dual role in calcium hydroxide-regulated DPSC behaviour. Incubation with calcium hydroxide promoted rapid proliferation and migration of DPSCs, while prolonged incubation negatively regulated osteogenic differentiation and mineralization.