Nerve growth factor (NGF)-conjugated electrospun nanostructures with topographical cues for neuronal differentiation of mesenchymal stem cells |
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| Authors: | Young Il Cho Ji Suk Choi Seo Young Jeong Hyuk Sang Yoo |
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| Affiliation: | 1. Department of Life and Nanopharmaceutical Science, Kyunghee University, Seoul, Republic of Korea;2. Department of Biomaterials Engineering, Kangwon National University, Chuncheon 200-701, Republic of Korea;3. Institute of Bioscience and Bioengineering, Kangwon National University, Republic of Korea;1. Department of Neurosurgery, The First Affiliated Hospital of Chongqing Medical University, Youyi Road 1, Yuzhong District, Chongqing 400016, China;2. Department of Chemistry, School of Life Sciences, University of Sussex, Brighton BN1 9QJ, United Kingdom;3. Department of Pharmacology, School of Pharmacy of Chongqing Medical University, Chongqing 400016, China;4. Department of Neurosurgery, Affiliated Hospital of Zunyi Medical College, Zunyi, Guizhou 563000, China;1. Center for Biomedical Engineering, Department of Medicine, Brigham and Women''s Hospital, Harvard Medical School, Cambridge, MA 02139, USA;2. Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA;3. Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA;4. David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA;5. Department of Electronics and Telecommunications, Politecnico di Torino, Torino 10129, Italy;6. School of Materials Science and Engineering, University of Science and Technology, Beijing, Beijing 100083, China;7. Center for Space Human Robotics, Italian Institute of Technology, Torino, Italy;1. Department of Orthopaedic Surgery, Duke University Medical Center, Durham, NC 27710, USA;2. Department of Biomedical Engineering, Duke University, Durham, NC 27708, USA;1. Foundation for Research and Technology-Hellas (F.O.R.T.H.), Institute of Electronic Structure and Laser (I.E.S.L.), Heraklion, Greece;2. Department of Biology, University of Crete, Heraklion, Greece;3. Department of Pharmacology, School of Medicine, University of Crete, Heraklion, Greece;4. Department of Physics, University of Crete, Heraklion, Greece;5. Department of Materials Science and Technology, University of Crete, Heraklion, Greece |
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| Abstract: | Mesenchymal stem cells (MSCs) were cultivated on the surface of nerve growth factor (NGF)-conjugated aligned nanofibrous meshes for neuronal differentiation. Amine-terminated poly(ethylene glycol) was conjugated to poly(ε-caprolactone) to prepare amine-functionalized block copolymers. The synthesized polymer was electrospun in a rotating drum to prepare aligned nanofibrous meshes. A nerve growth factor was chemically immobilized on the surface-exposed amine groups of the electrospun nanofibrous meshes in the aqueous phase. In vitro release profiles of the nerve growth factor were investigated for NGF-immobilized nanofibrous meshes. The conjugated nerve growth factor was not released for 7 days, while the growth factor physically adsorbed on the nanofibrous meshes showed an initial burst release. MSCs were cultivated on the NGF-conjugated nanofibrous meshes for 5 days, and total RNA was extracted from the cultivated cells. mRNA was extracted from cells for measuring expression levels of neuronal differentiation markers, including nestin, tubulin βIII and map2, in the cultivated stem cells. The conjugation of NGF significantly increased the expression levels of the marker proteins for neuron cells while physically adsorbed NGFs on nanofibrous meshes showed low expression of these marker genes. Furthermore, alignments of nanofibrous meshes clearly increased the expression levels of neuronal makers while the nanofibrous mesh without the topographical cue did not affect neuronal differentiation of the cultivated stem cells. Confocal microscopy revealed that the stem cells on the NGF-conjugated aligned nanofibrous meshes showed intense staining with antibodies against neuronal makers as well as elongated morphology compared to other groups. Thus, the NGF-conjugated nanofibrous meshes with topographical cues significantly increased the neuronal differentiation of mesenchymal stem cells in comparison to NGF-adsorbed nanofibrous meshes. |
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