3D Culture of Chondrocytes in Gelatin Hydrogels with Different Stiffness
Abstract
:1. Introduction
2. Materials and Methods
2.1. Synthesis of GelMA Macromers
2.2. 1H Nuclear Magnetic Resonance (NMR)
2.3. RGD Density Measurement
2.4. Preparation of GelMA Hydrogels
2.5. Mechanical Testing
2.6. Swelling Ratio Measurement
2.7. Measurement of Enzymatic Degradation of Hydrogels
2.8. Culture of Chondrocytes in Hydrogels
2.9. Cell Viability Assay
2.10. Cytoskeleton Actin Filament Staining
2.11. Histological Stainings
2.12. Quantification of DNA and Sulfated Glycosaminoglycan (sGAG)
2.13. Real-Time PCR Analysis
2.14. Statistical Analysis
3. Results and Discussion
3.1. Synthesis and Characterization of GelMA Macromers
3.2. RGD Density in Gelatin and GelMA Macromers
3.3. Young’s Modulus and Swelling Ratio of GelMA Hydrogels
3.4. Enzymatic Degradation of GelMA Hydrogels
3.5. Cell Viability in GelMA Hydrogels
3.6. Cytoskeleton Organization in GelMA Hydrogels
3.7. Histological Stainings
3.8. Quantification of DNA, Cartilaginous Matrices, and Gene Expression
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Sample | Feed Ratio of Gel (g)/MA (mL) | DoF (%) |
---|---|---|
Low GelMA | 5/0.2 | 25.8 ± 0.7 |
Medium GelMA | 5/1.0 | 52.5 ± 1.2 |
High GelMA | 5/5.0 | 91.7 ± 1.4 |
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Li, X.; Chen, S.; Li, J.; Wang, X.; Zhang, J.; Kawazoe, N.; Chen, G. 3D Culture of Chondrocytes in Gelatin Hydrogels with Different Stiffness. Polymers 2016, 8, 269. https://doi.org/10.3390/polym8080269
Li X, Chen S, Li J, Wang X, Zhang J, Kawazoe N, Chen G. 3D Culture of Chondrocytes in Gelatin Hydrogels with Different Stiffness. Polymers. 2016; 8(8):269. https://doi.org/10.3390/polym8080269
Chicago/Turabian StyleLi, Xiaomeng, Shangwu Chen, Jingchao Li, Xinlong Wang, Jing Zhang, Naoki Kawazoe, and Guoping Chen. 2016. "3D Culture of Chondrocytes in Gelatin Hydrogels with Different Stiffness" Polymers 8, no. 8: 269. https://doi.org/10.3390/polym8080269