EGFL7 Secreted By Human Bone Mesenchymal Stem Cells Promotes Osteoblast Differentiation Partly Via Downregulation Of Notch1-Hes1 Signaling Pathway

Stem Cell Rev Rep. 2023 May;19(4):968-982. doi: 10.1007/s12015-022-10503-z. Epub 2023 Jan 7.

Abstract

Background: Epidermal growth factor-like domain protein 7 (EGFL7) is a secreted protein that is differentially expressed in the bone microenvironment; however, the effect of EGFL7 on the osteogenesis of human bone marrow mesenchymal stem cells (hBMSCs) is largely unknown.

Methods: EGFL7 expression in the fracture microenvironment was analyzed based on the Gene Expression Omnibus (GEO) database. Knockdown of EGFL7 by small interfering RNA (siRNA) and in vitro stimulation with recombinant human EGFL7 (rhEGFL7) protein were used to assess alterations in downstream signaling and changes in the osteogenic differentiation and proliferation of hBMSCs. A γ-secretase inhibitor was used to further explore whether inhibition of Notch signaling rescued the osteogenic-inhibitory effect of EGFL7 knockdown in hBMSCs. A femoral defect model was established to verify the effect of recombinant mouse EGFL7 on bone healing in vivo.

Results: EGFL7 expression increased during hBMSC osteogenesis. Knockdown of EGFL7 impaired hBMSC osteogenesis and activated Notch1/NICD/Hes1 signaling. rhEGFL7 promoted hBMSC osteogenesis and downregulated Notch1 signaling. The osteoblast-inhibitory effect of EGFL7 knockdown was rescued by Notch1 signaling inhibition. Recombinant EGFL7 led to enhanced bone healing in mice with femoral defects.

Conclusions: EGFL7 promotes osteogenesis of hBMSCs partly via downregulation of Notch1 signaling.

Keywords: EGFL71; Hes3; Notch2; Osteogenesis5; hBMSCs4.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation / genetics
  • Cells, Cultured
  • Down-Regulation / genetics
  • Humans
  • Intercellular Signaling Peptides and Proteins / metabolism
  • Mesenchymal Stem Cells*
  • Mice
  • Osteoblasts / metabolism
  • Osteogenesis* / genetics
  • RNA, Small Interfering / metabolism
  • Receptor, Notch1 / genetics
  • Receptor, Notch1 / metabolism
  • Signal Transduction
  • Transcription Factor HES-1 / genetics
  • Transcription Factor HES-1 / metabolism
  • Transcription Factors / metabolism

Substances

  • Intercellular Signaling Peptides and Proteins
  • RNA, Small Interfering
  • Transcription Factors
  • HES1 protein, human
  • Transcription Factor HES-1
  • NOTCH1 protein, human
  • Receptor, Notch1
  • EGFL7 protein, human
  • Hes1 protein, mouse