Functionalization of Electrospun Nanofibers and Fiber Alignment Enhance Neural Stem Cell Proliferation and Neuronal Differentiation
Neural stem cells (NSCs) have the potential to generate the cells of the nervous system and, when cultured on nanofiber scaffolds, constitute a promising approach for neural tissue engineering. In this work, the impact of combining nanofiber alignment with functionalization of the electrospun poly-ε...
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Frontiers Media S.A.
2020-10-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2020.580135/full |
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| author | Miriam C. Amores de Sousa Carlos A. V. Rodrigues Inês A. F. Ferreira Maria Margarida Diogo Robert J. Linhardt Joaquim M. S. Cabral Frederico Castelo Ferreira |
| author_facet | Miriam C. Amores de Sousa Carlos A. V. Rodrigues Inês A. F. Ferreira Maria Margarida Diogo Robert J. Linhardt Joaquim M. S. Cabral Frederico Castelo Ferreira |
| author_sort | Miriam C. Amores de Sousa |
| collection | DOAJ |
| description | Neural stem cells (NSCs) have the potential to generate the cells of the nervous system and, when cultured on nanofiber scaffolds, constitute a promising approach for neural tissue engineering. In this work, the impact of combining nanofiber alignment with functionalization of the electrospun poly-ε-caprolactone (PCL) nanofibers with biological adhesion motifs on the culture of an NSC line (CGR8-NS) is evaluated. A five-rank scale for fiber density was introduced, and a 4.5 level, corresponding to 70–80% fiber density, was selected for NSC in vitro culture. Aligned nanofibers directed NSC distribution and, especially in the presence of laminin (PCL-LN) and the RGD-containing peptide GRGDSP (PCL-RGD), promoted higher cell elongation, quantified by the eccentricity and axis ratio. In situ differentiation resulted in relatively higher percentage of cells expressing Tuj1 in PCL-LN, as well as significantly longer neurite development (41.1 ± 1.0 μm) than PCL-RGD (32.0 ± 1.0 μm), pristine PCL (25.1 ± 1.2 μm), or PCL-RGD randomly oriented fibers (26.5 ± 1.4 μm), suggesting that the presence of LN enhances neuronal differentiation. This study demonstrates that aligned nanofibers, functionalized with RGD, perform as well as PCL-LN fibers in terms of cell adhesion and proliferation. The presence of the full LN protein improves neuronal differentiation outcomes, which may be important for the use of this system in tissue engineering applications. |
| first_indexed | 2024-12-11T16:55:29Z |
| format | Article |
| id | doaj.art-397aaf712650441d9944240eaa7e7022 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-12-11T16:55:29Z |
| publishDate | 2020-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-397aaf712650441d9944240eaa7e70222022-12-22T00:57:58ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852020-10-01810.3389/fbioe.2020.580135580135Functionalization of Electrospun Nanofibers and Fiber Alignment Enhance Neural Stem Cell Proliferation and Neuronal DifferentiationMiriam C. Amores de Sousa0Carlos A. V. Rodrigues1Inês A. F. Ferreira2Maria Margarida Diogo3Robert J. Linhardt4Joaquim M. S. Cabral5Frederico Castelo Ferreira6Institute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, PortugalInstitute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, PortugalInstitute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, PortugalInstitute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, PortugalCenter for Biotechnology and Interdisciplinary Studies, Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Troy, NY, United StatesInstitute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, PortugalInstitute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, PortugalNeural stem cells (NSCs) have the potential to generate the cells of the nervous system and, when cultured on nanofiber scaffolds, constitute a promising approach for neural tissue engineering. In this work, the impact of combining nanofiber alignment with functionalization of the electrospun poly-ε-caprolactone (PCL) nanofibers with biological adhesion motifs on the culture of an NSC line (CGR8-NS) is evaluated. A five-rank scale for fiber density was introduced, and a 4.5 level, corresponding to 70–80% fiber density, was selected for NSC in vitro culture. Aligned nanofibers directed NSC distribution and, especially in the presence of laminin (PCL-LN) and the RGD-containing peptide GRGDSP (PCL-RGD), promoted higher cell elongation, quantified by the eccentricity and axis ratio. In situ differentiation resulted in relatively higher percentage of cells expressing Tuj1 in PCL-LN, as well as significantly longer neurite development (41.1 ± 1.0 μm) than PCL-RGD (32.0 ± 1.0 μm), pristine PCL (25.1 ± 1.2 μm), or PCL-RGD randomly oriented fibers (26.5 ± 1.4 μm), suggesting that the presence of LN enhances neuronal differentiation. This study demonstrates that aligned nanofibers, functionalized with RGD, perform as well as PCL-LN fibers in terms of cell adhesion and proliferation. The presence of the full LN protein improves neuronal differentiation outcomes, which may be important for the use of this system in tissue engineering applications.https://www.frontiersin.org/articles/10.3389/fbioe.2020.580135/fullPCLfunctionalizationnanofiberselectrospinningneural stem cellslaminin |
| spellingShingle | Miriam C. Amores de Sousa Carlos A. V. Rodrigues Inês A. F. Ferreira Maria Margarida Diogo Robert J. Linhardt Joaquim M. S. Cabral Frederico Castelo Ferreira Functionalization of Electrospun Nanofibers and Fiber Alignment Enhance Neural Stem Cell Proliferation and Neuronal Differentiation Frontiers in Bioengineering and Biotechnology PCL functionalization nanofibers electrospinning neural stem cells laminin |
| title | Functionalization of Electrospun Nanofibers and Fiber Alignment Enhance Neural Stem Cell Proliferation and Neuronal Differentiation |
| title_full | Functionalization of Electrospun Nanofibers and Fiber Alignment Enhance Neural Stem Cell Proliferation and Neuronal Differentiation |
| title_fullStr | Functionalization of Electrospun Nanofibers and Fiber Alignment Enhance Neural Stem Cell Proliferation and Neuronal Differentiation |
| title_full_unstemmed | Functionalization of Electrospun Nanofibers and Fiber Alignment Enhance Neural Stem Cell Proliferation and Neuronal Differentiation |
| title_short | Functionalization of Electrospun Nanofibers and Fiber Alignment Enhance Neural Stem Cell Proliferation and Neuronal Differentiation |
| title_sort | functionalization of electrospun nanofibers and fiber alignment enhance neural stem cell proliferation and neuronal differentiation |
| topic | PCL functionalization nanofibers electrospinning neural stem cells laminin |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2020.580135/full |
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