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Osteogenic effects of rapamycin on bone marrow mesenchymal stem cells via inducing autophagy

Yifeng Xing1,2, Chaowei Liu1, Lin Zhou2

  • 1Fujian Key Laboratory of Oral Diseases & Fujian Provincial Engineering Research Center of Oral Biomaterial & Stomatological Key Lab of Fujian College and University, School and Hospital of Stomatology, Fujian Medical University, Fuzhou, China.

Journal of Orthopaedic Surgery and Research
|February 23, 2023
PubMed
Abstract

Insights

Autophagy decreases during bone marrow mesenchymal stem cell (BMSC) osteogenic differentiation. Rapamycin (RAPA) at appropriate concentrations activates autophagy, promoting BMSC osteogenic differentiation.

Area of Science:

  • Cell Biology
  • Stem Cell Biology
  • Biochemistry

Background:

  • Autophagy is crucial for stem cell functions like self-renewal and differentiation.
  • The specific role of autophagy in bone marrow mesenchymal stem cells (BMSCs) during osteogenic differentiation is not fully understood.

Purpose of the Study:

  • To investigate the relationship between autophagy and osteogenic differentiation in BMSCs.
  • To examine the effects of rapamycin (RAPA), an autophagy agonist, on BMSC osteogenic differentiation.

Main Methods:

  • Analyzed BMSC autophagy using western blotting, immunofluorescence, and RT-qPCR during osteogenic induction.
  • Assessed osteogenic differentiation via alizarin red staining, alkaline phosphatase assays, and quantification of key osteogenic markers.
  • Utilized rapamycin (RAPA) and 3-methyladenine to modulate autophagy and evaluate its impact on differentiation.

Main Results:

  • BMSC basal autophagy levels decreased during osteogenic differentiation, evidenced by reduced BECN1 and LC3-II/LC3-I ratio, and increased p62.
  • Rapamycin (RAPA) treatment increased autophagy; 2 nM RAPA promoted osteogenesis at early time points (days 7-14), while 5 nM RAPA inhibited it later (days 14-21).
  • Inhibiting autophagy with 3-methyladenine diminished the osteogenesis-promoting effect of RAPA.

Conclusions:

  • Basal autophagy declines progressively during BMSC osteogenic differentiation.
  • Autophagy activation, modulated by appropriate RAPA concentrations, can enhance BMSC osteogenic differentiation.
Keywords:
3-MethyladenineAutophagyBone marrow mesenchymal stem cellsOsteogenic differentiationRapamycin

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