The role and potential mechanism of p75NTR in mineralization via in vivo p75NTR knockout mice and in vitro ectomesenchymal stem cells

Cell Prolif. 2020 Feb;53(2):e12758. doi: 10.1111/cpr.12758. Epub 2020 Jan 10.

Abstract

Objective: The aim of this study is to investigate the role and potential mechanism of p75NTR in mineralization in vivo using p75NTR-knockout mice and in vitro using ectomesenchymal stem cells (EMSCs).

Materials and methods: Femur bone mass and daily incisor mineralization speed were assessed in an in vivo p75NTR-knockout mouse model. The molecular signatures alkaline phosphatase (ALP), collagen type 1 (Col1), melanoma-associated antigen (Mage)-D1, bone sialoprotein (BSP), osteocalcin (OCN), osteopontin (OPN), distal-less homeobox 1 (Dlx1) and Msh homeobox 1 (Msx1) were examined in vitro in EMSCs isolated from p75NTR+/+ and p75NTRExIII-/- mice.

Results: p75NTR-knockout mice were smaller in body size than heterozygous and wild-type mice. Micro-computed tomography and structural quantification showed that the osteogenic ability of p75NTRExIII -knockout mice was significantly decreased compared with that of wild-type mice (P < .05). Weaker ALP and alizarin red staining and reduced expression of ALP, Col1, Runx2, BSP, OCN and OPN were also observed in p75NTRExIII-/- EMSCs. Moreover, the distance between calcein fluorescence bands in p75NTRExIII -knockout mice was significantly smaller than that in wild type and heterozygous mice (P < .05), indicating the lower daily mineralization speed of incisors in p75NTRExIII -knockout mice. Further investigation revealed a positive correlation between p75NTR and Mage-D1, Dlx1, and Msx1.

Conclusion: p75NTR not only promotes osteogenic differentiation and tissue mineralization, but also shows a possible relationship with the circadian rhythm of dental hard tissue formation.

Keywords: ectomesenchymal stem cells; knockout mice; mineralization; p75 neurotrophin receptor; tooth.

MeSH terms

  • Alkaline Phosphatase / metabolism
  • Animals
  • Calcification, Physiologic / physiology
  • Cell Differentiation / physiology
  • Cells, Cultured
  • Collagen Type I / metabolism
  • Female
  • Integrin-Binding Sialoprotein / metabolism
  • Male
  • Mesenchymal Stem Cells / metabolism*
  • Mice
  • Mice, Knockout
  • Osteocalcin / metabolism
  • Osteogenesis / physiology
  • Osteopontin / metabolism
  • Receptors, Nerve Growth Factor / metabolism*

Substances

  • Collagen Type I
  • Integrin-Binding Sialoprotein
  • Receptors, Nerve Growth Factor
  • Ngfr protein, mouse
  • Osteocalcin
  • Osteopontin
  • Alkaline Phosphatase

Associated data

  • GENBANK/NM_001204203
  • GENBANK/X04142
  • GENBANK/NM_008318
  • GENBANK/NM_007431
  • GENBANK/NM_007732
  • GENBANK/NM_001146038
  • GENBANK/NM_008084