Exosomes from Microvascular Endothelial Cells under Mechanical Unloading Inhibit Osteogenic Differentiation via miR-92b-3p/ELK4 Axis
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture and In Vitro Differentiation
2.2. Two-Dimensional Clinorotation
2.3. Exosome Isolation and Characterization
2.4. Exosome Internalization
2.5. miRNA Loading into Exosomes
2.6. qRT–PCR
2.7. Western Blotting Analysis
2.8. Cell Transfection
2.9. ALP Activity Assay
2.10. ALP Staining
2.11. Luciferase Assay
2.12. Statistical Analysis
3. Results
3.1. Isolation, Identification, and Internalization of MVEC-Derived Exosomes
3.2. Exosomes Derived from MVECs Cultured under Mechanical Unloading Attenuate Osteoblast Differentiation
3.3. miR-92b-3p Is Elevated in MVEC-Derived Exosomes Cultured under Mechanical Unloading
3.4. miR-92b-3p Hinders the Osteogenic Differentiation of MC3T3-E1 Cells
3.5. Exosomes Derived from MVECs Cultured under Mechanical Unloading Inhibit Osteogenic Differentiation by Transferring miR-92b-3p
3.6. ELK4 Is a Direct Target of miR-92b-3p and Is Responsible for miR-92b-3p-Mediated Suppression of Osteogenic Differentiation in MC3T3-E1 Cells
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Zhang, X.; Zhang, L.; Xu, L.; Li, G.; Wang, K.; Xue, T.; Sun, Q.; Tang, H.; Cao, X.; Hu, Z.; et al. Exosomes from Microvascular Endothelial Cells under Mechanical Unloading Inhibit Osteogenic Differentiation via miR-92b-3p/ELK4 Axis. J. Pers. Med. 2022, 12, 2030. https://doi.org/10.3390/jpm12122030
Zhang X, Zhang L, Xu L, Li G, Wang K, Xue T, Sun Q, Tang H, Cao X, Hu Z, et al. Exosomes from Microvascular Endothelial Cells under Mechanical Unloading Inhibit Osteogenic Differentiation via miR-92b-3p/ELK4 Axis. Journal of Personalized Medicine. 2022; 12(12):2030. https://doi.org/10.3390/jpm12122030
Chicago/Turabian StyleZhang, Xiaoyan, Lijun Zhang, Liqun Xu, Gaozhi Li, Ke Wang, Tong Xue, Quan Sun, Hao Tang, Xinsheng Cao, Zebing Hu, and et al. 2022. "Exosomes from Microvascular Endothelial Cells under Mechanical Unloading Inhibit Osteogenic Differentiation via miR-92b-3p/ELK4 Axis" Journal of Personalized Medicine 12, no. 12: 2030. https://doi.org/10.3390/jpm12122030