The Modulation of Respiratory Epithelial Cell Differentiation by the Thickness of an Electrospun Poly-ε-Carprolactone Mesh Mimicking the Basement Membrane
Abstract
:1. Introduction
2. Results
2.1. Characteristics of Electrospun PCL Mesh
2.2. Viability and Barrier Function of Differentiated NHBE Cells during ALI
2.3. Generation of Airway Epithelium on the 6- and 80-Layer PCL Mesh
2.4. Hyperplastic Goblet Cells on the 80-Layer PCL Mesh Compared to on the 6-Layer PCL Mesh
2.5. The Expression of EMT Markers in Cells Differentiated on the 6- and 80-Layer PCL Meshes
2.6. Oxidative Stress-Related Signaling Pathway in NHBE Cells Differentiated on the 80-Layer PCL Mesh
3. Discussion
4. Materials and Methods
4.1. Fabrication of Electrospun PCL Mesh
4.2. Electrospun PCL Mesh Characterization and Tensile Strength Evaluation
4.3. Preperation of PCL Mesh and Expansion of Primary Airway Epithelial Cells
4.4. Differentiation of NHBE Cells by ALI
4.5. Cell Viability Assay
- At is the absorbance of NHBE cells on the membrane under ALI conditions;
- Ab is the absorbance of the membrane only under ALI conditions;
- At0 is the absorbance of NHBE cells on the membrane under ALI day 0;
- Ab0 is the absorbance of the membrane only under ALI day 0.
4.6. Trans-Epithelial Electrical Resistance (TEER)
4.7. Histological and Immunochemical Analysis
4.8. Alcian Blue Staining
4.9. Whole-Mount Immunostaining
4.10. Western Blot Analysis
4.11. Quantitative Real-Time Polymerase Chain Reaction (PCR)
4.12. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Unit: µm | ||
---|---|---|
Mean ± SD (Min, Max) | ||
6-Layer | 80-Layer | |
Fiber diameter | 0.23 ± 0.04 (0.18, 0.39) | 0.25 ± 0.02 (0.20, 0.28) |
Pore size | 0.59 ± 0.22 (0.27, 1.15) | 0.64 ± 0.41 (0.17, 1.81) |
Membrane thickness | 18.75 ± 1.92 (16.83, 20.67) | 256.83 ± 21.64 ** (235.19, 278.47) |
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Choi, S.Y.; Kim, H.J.; Hwang, S.; Park, J.; Park, J.; Lee, J.W.; Son, K.H. The Modulation of Respiratory Epithelial Cell Differentiation by the Thickness of an Electrospun Poly-ε-Carprolactone Mesh Mimicking the Basement Membrane. Int. J. Mol. Sci. 2024, 25, 6650. https://doi.org/10.3390/ijms25126650
Choi SY, Kim HJ, Hwang S, Park J, Park J, Lee JW, Son KH. The Modulation of Respiratory Epithelial Cell Differentiation by the Thickness of an Electrospun Poly-ε-Carprolactone Mesh Mimicking the Basement Membrane. International Journal of Molecular Sciences. 2024; 25(12):6650. https://doi.org/10.3390/ijms25126650
Chicago/Turabian StyleChoi, Seon Young, Hyun Joo Kim, Soyoung Hwang, Jangho Park, Jungkyu Park, Jin Woo Lee, and Kuk Hui Son. 2024. "The Modulation of Respiratory Epithelial Cell Differentiation by the Thickness of an Electrospun Poly-ε-Carprolactone Mesh Mimicking the Basement Membrane" International Journal of Molecular Sciences 25, no. 12: 6650. https://doi.org/10.3390/ijms25126650