The Paracrine Effect of Hyaluronic Acid-Treated Endothelial Cells Promotes BMP-2-Mediated Osteogenesis
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture and Chemicals
2.2. In Vitro Tube Formation Assay
2.3. Angiogenic Differentiation of HUVECs
2.4. HUVEC Conditioned Medium Preparation and BMSC Culture
2.5. Osteogenic Differentiation Assay
2.5.1. Alkaline Phosphatase (ALP) Staining and ALP Activity Assay
2.5.2. Alizarin Red Staining
2.5.3. mRNA Expression of Osteogenic Markers
2.6. RNA Isolation and Real-Time Quantitative PCR (RT-qPCR) Analysis
2.7. Animals, Anesthesia, and Surgery
2.8. Micro-CT Analysis
2.9. Masson’s Trichrome Staining
2.10. Statistical Analysis
3. Results
3.1. HA Promotes the Angiogenic Differentiation of HUVECs
3.2. HA Inhibits, but CM from HA-Treated HUVECs Promotes, BMP-2-Mediated Osteogenic Differentiation of BMSCs
3.3. HA + BMP-2 Treatment Upregulates Angiogenesis-Related Marker Genes’ Expression in HUVECs
3.4. HA + BMP-2 Synergistically Enhanced In Vivo Bone Regeneration
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Gene | Primer Sequence | Product Length (bp) |
---|---|---|
Mus GAPDH | F: GTGAAGGTCGGTGTGAACGG | 227 |
R: TCCTGGAAGATGGTGATGGG | ||
Mus Collagen I | F: ATGCCGCGACCTCAAGATG | 140 |
R: TGAGGCACAGACGGCTGAGTA | ||
Mus RUNX2 | F: TGAGGCACAGACGGCTGAGTA | 126 |
R: CACTGGCGGTGCAACAAGA | ||
Mus OPN | F: ACCATGCAGAGAGCGAGGATT | 91 |
R: GGGACATCGACTGTAGGGACG | ||
Mus ALP | F: TGCCTACTTGTGTGGCGTGAA | 159 |
R: TCACCCGAGTGGTAGTCAVAATG | ||
Homo GAPDH | F: GAAGGTGAAGGTCGGAGTCA | 172 |
R: GAAGATGGTGATGGGATTTC | ||
Homo CD31 | F: CTCCAGACTCCACCACCTTAC | 243 |
R: GAACTTTGCCTATTTCTTACCA | ||
Homo VEGF | F: GGAGGCAGAGAAAAGAGAAAGTGT | 175 |
R: TAAGAGAGCAAGAGAGAGCAAAAGA | ||
Homo bFGF | F: AGCCAGGTAACGGTTAGCACA | 91 |
R: GAAGAGCGACCCTCACATCAA |
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Tong, X.; Chen, J.; Wang, R.; Hou, D.; Wu, G.; Liu, C.; Pathak, J.L. The Paracrine Effect of Hyaluronic Acid-Treated Endothelial Cells Promotes BMP-2-Mediated Osteogenesis. Bioengineering 2023, 10, 1227. https://doi.org/10.3390/bioengineering10101227
Tong X, Chen J, Wang R, Hou D, Wu G, Liu C, Pathak JL. The Paracrine Effect of Hyaluronic Acid-Treated Endothelial Cells Promotes BMP-2-Mediated Osteogenesis. Bioengineering. 2023; 10(10):1227. https://doi.org/10.3390/bioengineering10101227
Chicago/Turabian StyleTong, Xiaojie, Jin Chen, Renqin Wang, Dan Hou, Gang Wu, Chang Liu, and Janak Lal Pathak. 2023. "The Paracrine Effect of Hyaluronic Acid-Treated Endothelial Cells Promotes BMP-2-Mediated Osteogenesis" Bioengineering 10, no. 10: 1227. https://doi.org/10.3390/bioengineering10101227