Single-cell analysis of ecto-mesenchymal cells derived from the neural crest reveals the regulatory role of P75NTR in odontogenic and osteogenic differentiation

Abstract

Background

The aim to this study was to explore the effect of p75NTR on the mineralization and development of maxillofacial processes. Moreover, we tried to elaborate the potential mechanism of p75NTR’s effect on the odontogenic or osteogenic differentiation ability of neural crest cells (s).

Methods

We used gene p75NTR knockout (p75NTR^−/−) mice and wildtype (p75NTR^+/+) mice embryos as a model. And We isolated p75NTR^−/− and p75NTR^+/+ ectodermal mesenchymal stem cells (EMSCs) from the embryonic maxillofacial process of the same pregnant p75NTR^+/− mice. The potential mechanism of p75NTR’s effect on the odontogenic or osteogenic differentiation ability of EMSCs was analyzed by single cell RNA sequence(scRNA-seq). The results were further verified by mineralize induction and wound-healing Assay.

Results

We found the deletion of p75NTR could impact the communications between different cell types, influence differentiation and suppress odontogenesis via regulation networks of cytokines that associating with migration, proliferation and differentiation. The volcano plot characterized the distribution of differentially expressed genes (DEGs) in the selection at threshold level. p75NTR deletion suppressed EMSCs migration, proliferation and odonto/osteogenic differentiation in vitro.

Conclusions

This study provides a comprehensive analysis of the cellular fate of EMSCs in the maxillofacial process. p75NTR plays a crucial role in initiating hard tissue development and regulates the odontogenic and osteogenic differentiation of EMSCs during embryonic development.