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Osteogenic effects of rapamycin on bone marrow mesenchymal stem cells via inducing autophagy.
- Source :
-
Journal of Orthopaedic Surgery & Research . 2/22/2023, Vol. 18 Issue 1, p1-13. 13p. - Publication Year :
- 2023
-
Abstract
- Background: While autophagy is essential for stem cells' self-renewal and differentiation, its effect on bone marrow mesenchymal stem cells (BMSCs) remains unclear. This study aimed to investigate the interaction between autophagy and osteogenic differentiation using rapamycin (RAPA), a classical autophagy agonist with osteo-regulatory effects. Methods: Rat BMSC's autophagy was analyzed after osteoinduction (0, 7, 14, and 21 d) by western blotting, immunofluorescence, and real-time quantitative polymerase chain reaction (RT-qPCR). In addition, we evaluated osteogenic differentiation using alizarin red staining, alkaline phosphatase assays, and RT-qPCR/Western blotting quantification of bone sialoprotein, type 1 collagen, alkaline phosphatase, osteopontin, and Runt-related transcription factor 2 mRNA and protein levels. Results: The BMSC's basal autophagy level gradually decreased during osteogenic differentiation with a decrease in BECN1 level and the lipidated (LC3-II) to unlipidated (LC3-I) microtubule-associated protein 1 light chain 3 ratio and an increase in the expression of selective autophagic target p62. In contrast, it increased with increasing RAPA concentration. Furthermore, while 2 nM RAPA promoted BMSC osteogenic differentiation on days 7 and 14, 5 nM RAPA inhibited osteogenesis on days 14 and 21. Inhibition of autophagy by the inhibitor 3-methyladenine could impair RAPA's osteogenesis-enhancing effect on BMSCs. Conclusions: The BMSC's basal autophagy level decreased over time during osteogenic differentiation. However, an appropriate RAPA concentration promoted BMSC osteogenic differentiation via autophagy activation. [ABSTRACT FROM AUTHOR]
- Subjects :
- *REVERSE transcriptase polymerase chain reaction
*ALKALINE phosphatase
*CELL differentiation
*PROTEINS
*BONE growth
*RAPAMYCIN
*CELL culture
*AUTOPHAGY
*ANIMAL experimentation
*WESTERN immunoblotting
*QUANTITATIVE research
*CELL receptors
*RATS
*FLUORESCENT antibody technique
*MESSENGER RNA
*RESEARCH funding
*BONE marrow
*MESENCHYMAL stem cells
Subjects
Details
- Language :
- English
- ISSN :
- 1749799X
- Volume :
- 18
- Issue :
- 1
- Database :
- Academic Search Index
- Journal :
- Journal of Orthopaedic Surgery & Research
- Publication Type :
- Academic Journal
- Accession number :
- 162032405
- Full Text :
- https://doi.org/10.1186/s13018-023-03616-9