Modeling In Vitro Osteoarthritis Phenotypes in a Vascularized Bone Model Based on a Bone-Marrow Derived Mesenchymal Cell Line and Endothelial Cells
Abstract
:1. Introduction
2. Results
2.1. Generation of 3D Constructs Made of MSODs and HUVECs Co-Cultured in gelMA
2.2. HUVECs Enhanced Osteogenic Differentiation of MSODs in the Co-Culture System
2.3. MSOD Helped Stabilization of the HUVEC Network Formation after 2 Weeks of Culture
2.4. OA Phenotype Induction though Administration of an Inflammatory Cocktail
2.5. Effects of Conditioned Medium from OA Cartilage-on-Chip
3. Discussion
4. Materials and Methods
4.1. Generation of the Constructs and OA Induction
4.2. Cell Viability
4.3. Quantitative RT-PCR Analysis
4.4. Alizarin Red Staining and Quantification
4.5. Immunofluorescence
4.6. Quantification of Alkaline Phosphate (ALP) Activity and Calcium Content in the Surnatants
4.7. Angiogenesis Analysis
4.8. OA Cartilage-on-Chip Model Setup
4.9. Microfluidic Devices Fabrication
4.10. Supernatant Analyses
4.11. Statistical Analysis
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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Gene | Assay on Demand Ref. No. |
---|---|
GAPDH | Hs02758991_g1 |
ALP | Hs01029144_m1 |
OSX | Hs00541729_m1 |
BSP | Hs00173720_m1 |
OPN | Hs00959010_m1 |
COL1A1 | Hs01097664_m1 |
COL1A2 | Hs01028960_m1 |
OSC | Hs01587814_g1 |
VEGF | Hs00900055_m1 |
WNT5a | Hs00998537_m1 |
ADAMTS5 | Hs00199841_m1 |
MMP-13 | Hs00233992_m1 |
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Pirosa, A.; Tankus, E.B.; Mainardi, A.; Occhetta, P.; Dönges, L.; Baum, C.; Rasponi, M.; Martin, I.; Barbero, A. Modeling In Vitro Osteoarthritis Phenotypes in a Vascularized Bone Model Based on a Bone-Marrow Derived Mesenchymal Cell Line and Endothelial Cells. Int. J. Mol. Sci. 2021, 22, 9581. https://doi.org/10.3390/ijms22179581
Pirosa A, Tankus EB, Mainardi A, Occhetta P, Dönges L, Baum C, Rasponi M, Martin I, Barbero A. Modeling In Vitro Osteoarthritis Phenotypes in a Vascularized Bone Model Based on a Bone-Marrow Derived Mesenchymal Cell Line and Endothelial Cells. International Journal of Molecular Sciences. 2021; 22(17):9581. https://doi.org/10.3390/ijms22179581
Chicago/Turabian StylePirosa, Alessandro, Esma Bahar Tankus, Andrea Mainardi, Paola Occhetta, Laura Dönges, Cornelia Baum, Marco Rasponi, Ivan Martin, and Andrea Barbero. 2021. "Modeling In Vitro Osteoarthritis Phenotypes in a Vascularized Bone Model Based on a Bone-Marrow Derived Mesenchymal Cell Line and Endothelial Cells" International Journal of Molecular Sciences 22, no. 17: 9581. https://doi.org/10.3390/ijms22179581