Bone Morphogenetic Protein-2 Signaling in the Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells Induced by Pulsed Electromagnetic Fields
Abstract
:1. Introduction
2. Results
2.1. Dose-Response Effects of BMP2 on hMSCs Osteogenic Differentiation
2.2. Effects of PEMF Exposure and BMP2 on hMSCs Osteogenic Differentiation
2.3. Effects of PEMF Exposure and BMP2 on Gene Expression of BMPs and BMP Receptors During hMSCs Osteogenic Differentiation
2.4. Immunoblotting
2.5. Effects of Dorsomorphin and SB203580 on Osteogenic Differentiation
3. Discussion
4. Materials and Methods
4.1. Cell Cultures
4.2. Osteogenic Differentiation and Cell Treatments
- OM + BMP2;
- OM + PEMFs;
- OM + PEMFs + BMP2.
4.3. PEMFs and Exposure Conditions
4.4. SMAD and MAPK Signaling Inhibition
4.5. Osteogenic Markers
4.5.1. Alkaline Phosphatase (ALP) Activity
4.5.2. Osteocalcin (OC) Levels
4.6. Alizarin Red S (ARS) Staining and Quantification
4.7. Quantitative Real-Time PCR (qPCR)
4.8. Western Blotting and Densitometric Analysis
4.9. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
PEMFs | Pulsed electromagnetic fields |
MSCs | Mesenchymal stem cells |
BMP2 | Bone morphogenetic protein-2 |
hMSCs | Human MSCs |
EMF | Electromagnetic field |
BMPs | Bone morphogenetic proteins |
TGF-β | Transforming growth factors beta |
CHO | Chinese hamster ovary |
rhBMP2 | Recombinant human BMP-2 |
MAPKs | Mitogen-activated protein kinases |
OSX | Osterix |
DLX | Distal-less homeobox |
MSCBM | Mesenchymal stem cell basal medium |
ALP | Alkaline phosphatase |
OC | Osteocalcin |
OM | Osteogenic medium |
DM | Dorsomorphin |
p-NP | p-nitrophenylphosphate |
DNA | Deoxyribonucleic acid |
ARS | Alizarin red Staining |
qPCR | Quantitative PCR |
GUSB | Glucuronidase beta |
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Martini, F.; Pellati, A.; Mazzoni, E.; Salati, S.; Caruso, G.; Contartese, D.; De Mattei, M. Bone Morphogenetic Protein-2 Signaling in the Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells Induced by Pulsed Electromagnetic Fields. Int. J. Mol. Sci. 2020, 21, 2104. https://doi.org/10.3390/ijms21062104
Martini F, Pellati A, Mazzoni E, Salati S, Caruso G, Contartese D, De Mattei M. Bone Morphogenetic Protein-2 Signaling in the Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells Induced by Pulsed Electromagnetic Fields. International Journal of Molecular Sciences. 2020; 21(6):2104. https://doi.org/10.3390/ijms21062104
Chicago/Turabian StyleMartini, Fernanda, Agnese Pellati, Elisa Mazzoni, Simona Salati, Gaetano Caruso, Deyanira Contartese, and Monica De Mattei. 2020. "Bone Morphogenetic Protein-2 Signaling in the Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells Induced by Pulsed Electromagnetic Fields" International Journal of Molecular Sciences 21, no. 6: 2104. https://doi.org/10.3390/ijms21062104