Lipopolysaccharide-Preconditioned Periodontal Ligament Stem Cells Induce M1 Polarization of Macrophages through Extracellular Vesicles
Abstract
:1. Introduction
2. Results
2.1. Characterization of Human Periodontal Ligament Stem Cells (PDLSCs)
2.2. Effect of the Supernatant from PDLSCs on Macrophage Polarization
2.3. LPS-Preconditioned PDLSCs Accentuate the M1 Polarization of IFN-γ Treated Macrophages
2.4. M1 Polarization Was Not Mediated by Free (Soluble) Cytokines from PDLSCs
2.5. Isolation of EVs from PDLSCs and Their Characterization
2.6. EVs from LPS-Preconditioned PDLSCs Synergize with the Action of IFN-γ on Macrophages
2.7. DNase I Treatment Abolishes M1 Polarization Effect of EVs
3. Discussion
4. Materials and Methods
4.1. Cell Culture and Reagents
4.2. Flow Cytometry
4.3. Osteogenic Differentiation of PDLSCs
4.4. Quantitative Real-Time Reverse Transcription PCR (RT-qPCR)
4.5. Induction of M1 and M2 Phenotype of THP-1 Cells
4.6. Isolation of EVs from PDLSCs
4.7. Nano-Particle Tracking Analysis
4.8. Labeling and Tracking of the Entry of EVs
4.9. Remove of Endotoxin from EVs
4.10. TNF-α ELISA
4.11. Statistical Analyzes
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Kang, H.; Lee, M.-J.; Park, S.J.; Lee, M.-S. Lipopolysaccharide-Preconditioned Periodontal Ligament Stem Cells Induce M1 Polarization of Macrophages through Extracellular Vesicles. Int. J. Mol. Sci. 2018, 19, 3843. https://doi.org/10.3390/ijms19123843
Kang H, Lee M-J, Park SJ, Lee M-S. Lipopolysaccharide-Preconditioned Periodontal Ligament Stem Cells Induce M1 Polarization of Macrophages through Extracellular Vesicles. International Journal of Molecular Sciences. 2018; 19(12):3843. https://doi.org/10.3390/ijms19123843
Chicago/Turabian StyleKang, Hyejong, Myung-Ju Lee, Sang June Park, and Myung-Shin Lee. 2018. "Lipopolysaccharide-Preconditioned Periodontal Ligament Stem Cells Induce M1 Polarization of Macrophages through Extracellular Vesicles" International Journal of Molecular Sciences 19, no. 12: 3843. https://doi.org/10.3390/ijms19123843