PLA Electrospun Scaffolds for Three-Dimensional Triple-Negative Breast Cancer Cell Culture
"> Figure 1
<p>DSC curve of poly(lactic acid) (PLA).</p> "> Figure 2
<p>TGA curve of PLA.</p> "> Figure 3
<p>Features of 12% and 15% ES-PLA scaffolds. Images taken from the top and bottom sides were used to calculate (<b>a</b>) fiber diameter, (<b>b</b>) surface porosity, and (<b>c</b>) pore diameter. Results are shown as average ± SE. SF: small fibers; LF: large fibers.</p> "> Figure 4
<p>Cell proliferation analysis for MDA-MB-231 cells cultured in 3D using 12% PLA scaffolds (black bars) or 15% PLA scaffolds (grey bars) over three days (left) or six days (right). MDA-MB-231 cells cultured in 2D were used as a control. Experiments were performed at least three times.</p> "> Figure 5
<p>EGFR and STAT3 genes expression and protein levels. (<b>a</b>) EGFR and STAT3 gene expression of MDA-MB-231 cells cultured in 2D (black bars) and in 3D (grey bars) using 15% PLA scaffolds over three days (left) or six days (right). (<b>b</b>) Quantification of protein levels (EGFR and STAT3) obtained by Western blot analyses (right panel) of MDA-MB-231 cells cultured in 2D (black bars) and in 3D (grey bars) using 15% PLA scaffolds over three days (left) or six days (right). Results are expressed as the ratio of activated protein (p-Protein) vs. total levels of protein (t-Protein). Experiments were performed at least three times. * (<span class="html-italic">p</span> < 0.05) indicates levels of statistical significance. For an easier interpretation, the value obtained for cells cultured in 2D was adjusted to 1 and that of the 3D-cultured cells was calculated based on this.</p> "> Figure 6
<p>Mammosphere-forming Index (MFI) of MDA-MB-231 TNBC cells after monolayer (2D) or 15% PLA scaffolds (3D) culture. Experiments were performed at least three times.</p> "> Figure 7
<p>Epithelial-to-mesenchymal transition (EMT)-related and stemness gene expression. Stemness (Sox2) and EMT-related (Snail, E-cadherin, and Vimentin) gene expression of MDA-MB-231 culture in 2D (black bars) and in 3D (grey bars) using 15% PLA ES scaffolds over three days (left) and six days (right). Experiments were performed at least three times. For an easier interpretation, the value obtained for cells cultured in 2D was adjusted to 1 and that of the 3D-cultured cells was calculated based on this. * (<span class="html-italic">p</span> < 0.05) indicates levels of statistical significance.</p> ">
Abstract
:1. Introduction
2. Materials and Methods
2.1. Polymer
2.2. Polymer Thermal Analysis
2.3. Electrospun Scaffolds Manufacturing
2.4. Scaffold Physical Characterization
2.5. Cell Line and Culture Conditions
2.6. Three-Dimensional Cell Culture
2.7. Cell Proliferation Assay
2.8. Quantitative Real-Time PCR Analysis
2.9. Protein Analysis
2.10. Mammosphere-Forming Assay
2.11. Statistical Analysis
3. Results
3.1. Thermal Characterization of PLA
3.2. Physical Characterization of ES-PLA Scaffolds
3.3. High Proliferation Rates are Obtained in Both 12% and 15% ES-PLA Scaffolds
3.4. EGFR and STAT3 are Altered in 3D-Cultured Cells
3.5. ES-PLA Scaffolds Do Not Increase BCSC Population
4. Discussion
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Molecular Weight (g/mol) | Melting Point (°C) | Glass Transition (°C) | Young’s Modulus (MPa) | Strain at Break (%) | Degradation Time (Months) |
---|---|---|---|---|---|
30,000 | 173–178 | 60–65 | 108 | 3.5% | 12 |
Gene | Forward sequence (5′-3′) | Reverse sequence (5′-3′) |
---|---|---|
SNAIL | GCTGCAGGACTCTAATCCAGA | ATCTCCGGAGGTGGGATG |
SOx2 | AACCCCAAGATGCACAACTC | GCTTAGCCTCGTCGATGAAC |
E-cadh | TGGAGGAATTCTTGCTTTGC | CGCTCTCCTCCGAAGAAAC |
VIM | TGGTCTAACGGTTTCCCCTA | GACCTCGGAGCGAGAGTG |
EGFR | CATGTCGATGGACTTCCAGA | GGGACAGCTTGGATCACACT |
STAT3 | CACCTTCAGGATGTCCGGAA | ATCCTGGAGATTCTCTACCACTTTCA |
GAPDH | TCTTCCAGGAGCGAGATC | CAGAGATGATGACCCTTTTG |
Scaffold trait | 12% | 15% |
---|---|---|
Weight (mg) | 5.83 ± 0.63 | 6.23 ± 0.44 |
Thickness (µm) | 68.33 ± 6.89 | 68.00 ± 3.51 |
12% PLA | Side | Magnification | ||
200× | 1500× | 5000× | ||
Top | ||||
Bottom | ||||
15% PLA | Top | |||
Bottom | ||||
Scale bars: 150 µm | Scale bars: 20 µm | Scale bars: 6 µm |
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Polonio-Alcalá, E.; Rabionet, M.; Gallardo, X.; Angelats, D.; Ciurana, J.; Ruiz-Martínez, S.; Puig, T. PLA Electrospun Scaffolds for Three-Dimensional Triple-Negative Breast Cancer Cell Culture. Polymers 2019, 11, 916. https://doi.org/10.3390/polym11050916
Polonio-Alcalá E, Rabionet M, Gallardo X, Angelats D, Ciurana J, Ruiz-Martínez S, Puig T. PLA Electrospun Scaffolds for Three-Dimensional Triple-Negative Breast Cancer Cell Culture. Polymers. 2019; 11(5):916. https://doi.org/10.3390/polym11050916
Chicago/Turabian StylePolonio-Alcalá, Emma, Marc Rabionet, Xavier Gallardo, David Angelats, Joaquim Ciurana, Santiago Ruiz-Martínez, and Teresa Puig. 2019. "PLA Electrospun Scaffolds for Three-Dimensional Triple-Negative Breast Cancer Cell Culture" Polymers 11, no. 5: 916. https://doi.org/10.3390/polym11050916