Primary Human Ligament Fibroblast Adhesion and Growth on 3D-Printed Scaffolds for Tissue Engineering Applications
Abstract
:1. Introduction
2. Materials and Methods
2.1. Scaffold Fabrication
2.2. Surface Coating of the Scaffolds
2.3. Isolation and Culture of Primary Human Ligament Fibroblasts
2.4. Cell Seeding and Culture on the Scaffolds
2.5. Mechanical Testing
2.6. Scaffold Geometry
2.7. Surface Morphology
2.8. Gene Expression
2.9. Cell Viability and Proliferation
2.10. Immunofluorescence and Protein Analyses
2.11. Statistical Analysis
3. Results
3.1. Mechanical Testing and Scaffold Characterization
3.2. Cell Isolation, Characterization, and Scaffold Adhesion
3.3. Cell Viability, Proliferation, and DNA Quantification
3.4. Ligamentogenic Matrix Formation
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Target Gene | Product Length (bp) | Genbank’s Gene ID |
---|---|---|
GAPDH | 1978 | NM_001170721 |
COLIA1 | 462 | NM_001102599 |
COL3A1 | ||
TN-C | 1065 | AL445645.10 |
SCX | 1930 | NG_053063.1 |
Target Gene | Forward Primer (5′—3′) | Reverse Primer (3′—5′) |
---|---|---|
GAPDH | TCCCTGAGCTGAACGGGAAG | GGAGGAGTGGGTGTCGCTGT |
COLIA1 | GGTGATGCTGGTCCTGTTG | CATCGTGAGCCTTCTCTTGAG |
COL3A1 | CCCAGAACATCACATATCAC | CAAGAGGAACACATATGGAG |
TN-C | ACCGTCTCTTCCGTCACTTCT | AACAACTTAGGACAATGCGTCT |
SCX | AGAACACCCAGCCCAAACA | TCCTTGCTCAACTTTCTCTGGT |
Property | Value |
---|---|
Dry mass (mg) | 88.3 ± 0.7 |
SV/RV (%) | 60.7 ± 1.0 |
SS (mm2) | 159.5 ± 2.2 |
SF.Th (μm) | 479.9 ± 10.7 |
SF. Sp (μm) | 271.7 ± 4.6 |
Po. Tot (%) | 39.3 ± 1.0 |
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Lacombe, J.-G.; Cooke, M.E.; Park, H.; Alshammari, S.M.; Gawri, R.; Nazhat, S.N.; Martineau, P.A.; Rosenzweig, D.H. Primary Human Ligament Fibroblast Adhesion and Growth on 3D-Printed Scaffolds for Tissue Engineering Applications. Surgeries 2023, 4, 196-211. https://doi.org/10.3390/surgeries4020021
Lacombe J-G, Cooke ME, Park H, Alshammari SM, Gawri R, Nazhat SN, Martineau PA, Rosenzweig DH. Primary Human Ligament Fibroblast Adhesion and Growth on 3D-Printed Scaffolds for Tissue Engineering Applications. Surgeries. 2023; 4(2):196-211. https://doi.org/10.3390/surgeries4020021
Chicago/Turabian StyleLacombe, Jean-Gabriel, Megan E. Cooke, Hyeree Park, Suliman Mohammed Alshammari, Rahul Gawri, Showan N. Nazhat, Paul A. Martineau, and Derek H. Rosenzweig. 2023. "Primary Human Ligament Fibroblast Adhesion and Growth on 3D-Printed Scaffolds for Tissue Engineering Applications" Surgeries 4, no. 2: 196-211. https://doi.org/10.3390/surgeries4020021