LNGFR targets the Wnt/\u03b2-catenin pathway and promotes the osteogenic differentiation in rat ectomesenchymal stem cells

Gang Li,Yingying Wang,Luchuan Liu,Manzhu Zhao,Junyu Liu,Xiujie Wen,Kun Yang,Yong Xiao

doi:10.1038/s41598-017-11555-9

Abstract

Considerable evidence has shown that the Wnt/β-catenin pathway is involved in osteogenic differentiation in various stem cells. However, the role of Wnt/β-catenin pathway in regulating the osteogenic differentiation of rat ectomesenchymal stem cells (EMSCs), which are considered to be the progenitors of dental mesenchymal stem cells, remains unknown. In this study, we demonstrated that nuclear β-catenin was upregulated during EMSC osteogenic differentiation. The Wnt signalling inhibitor IWR-1-endo inhibited EMSC osteogenic differentiation, while the Wnt signalling agonist SKL2001 promoted it. Moreover, nuclear β-catenin was further upregulated by the overexpression of low-affinity nerve growth factor receptor (LNGFR) during EMSC osteogenic differentiation. Further experiments demonstrated that LNGFR overexpression enhanced EMSC osteogenic differentiation, while LNGFR silencing decreased it. Additionally, IWR-1-endo attenuated LNGFR-enhanced EMSC osteogenic differentiation. Collectively, our data reveal that LNGFR targets the Wnt/β-catenin pathway and positively regulates EMSC osteogenic differentiation, suggesting that Wnt/β-catenin pathway may be involved in the development of teeth and that the targeting Wnt/β-catenin pathway may have great potential for applications in dental tissue engineering regeneration.

Highlights

Ectomesenchymal stem cells (EMSCs), which are isolated from rat embryonic facial processes, are derived from the cranial neural crest[1]
We demonstrate that nuclear β-catenin levels are higher in ectomesenchymal stem cells (EMSCs) separated at E19.5d compared to EMSCs separated at E12.5d and upregulated during EMSC osteogenic differentiation
Further experiments showed that nuclear β-catenin is further upregulated by low-affinity nerve growth factor receptor (LNGFR) overexpression during EMSC osteogenic differentiation

Summary

Introduction

Ectomesenchymal stem cells (EMSCs), which are isolated from rat embryonic facial processes, are derived from the cranial neural crest[1]. Mao et al reported that the canonical Wnt signalling inhibitor dickkopf[1] repressed the mnHA bioceramics-induced upregulation of ALP activity and osteogenic and cementogenic gene expression in human periodontal ligament stem cells[11]. Yang et al reported that Wnt3a, a representative canonical member of the Wnt family of ligands, upregulates the expression of β-catenin and enhances the osteogenic differentiation of dental follicle cells, while dickkopf[1], a Wnt/β-catenin signalling inhibitor, decreases the expression of β-catenin and attenuates the osteogenic differentiation of dental follicle cells[13] It remains unknown whether the Wnt/β-catenin pathway participates in the osteogenic differentiation of EMSCs, which are the progenitor cells of dental mesenchymal stem cells. Our data revealed that LNGFR targets the Wnt/β-catenin pathway and positively regulates EMSC osteogenic differentiation, suggesting that the Wnt/β-catenin pathway may be involved in tooth development and that targeting the Wnt/β-catenin pathway may have great potential for applications in dental tissue engineering regeneration

Methods

Results

Conclusion