Introduction: An early substantial loss of basal forebrain cholinergic neurons (BFCNs) is a common property of Alzheimer’s disease and the degeneration of functional BFCNs is related to learning and memory deficits. As a biocompatible and conductive scaffold for growth of neural stem cells, three-dimensional graphene foam (3D-GF) supports applications in tissue engineering and regenerative medicine. Although its effects on differentiation have been demonstrated, the effect of 3D-GF scaffold on the generation of BFCNs still remains unknown. Methods: In this study, we used 3D-GF as a culture substrate for neural progenitor cells (NPCs) and demonstrated that this scaffold material promotes the differentiation of BFCNs while maintaining excellent cell viability and proliferation. Results: Immunofluorescence analysis, real-time polymerase chain reaction, Western blotting, and ELISA revealed that the proportion of BFCNs at 21 days of differentiation reached approximately 30.5% on 3D-GF compared with TCPS group that only presented 9.7%. Furthermore, a cell adhesion study suggested that 3D-GF scaffold enhances the expression of adhesion proteins including vinculin, integrin, and N-cadherin. These findings indicate that 3D-GF scaffold materials are preferable candidates for the differentiation of BFCNs from NPCs. Conclusions: These results suggest new opportunities for the application of 3D-GF scaffold as a neural scaffold for cholinergic neurons therapies based on NPCs.

Antequera D, Portero A, Bolos M, Orive G, Hernandez RM, Pedraz JL, et al. Encapsulated VEGF-secreting cells enhance proliferation of neuronal progenitors in the hippocampus of AβPP/Ps1 mice. J Alzheimers Dis. 2012;29(1):187–200.
Bissonnette CJ, Lyass L, Bhattacharyya BJ, Belmadani A, Miller RJ, Kessler JA. The controlled generation of functional basal forebrain cholinergic neurons from human embryonic stem cells. Stem Cell. 2011;29(5):802–11.
Chao TI, Xiang S, Chen CS, Chin WC, Nelson AJ, Wang C, et al. Carbon nanotubes promote neuron differentiation from human embryonic stem cells. Biochem Biophys Res Commun. 2009;384(4):426–30.
Chen Z, Ren W, Gao L, Liu B, Pei S, Cheng HM. Three-dimensional flexible and conductive interconnected graphene networks grown by chemical vapour deposition. Nat Mater. 2011;10(6):424–8.
Critchley DR, Holt MR, Barry ST, Priddle H, Hemmings L, Norman J. Integrin-mediated cell adhesion: the cytoskeletal connection. Biochem Soc Symp. 1999;65:79–99.
Crowder SW, Prasai D, Rath R, Balikov DA, Bae H, Bolotin KI, et al. Three-dimensional graphene foams promote osteogenic differentiation of human mesenchymal stem cells. Nanoscale. 2013;5(10):4171–6.
Cusimano M, Biziato D, Brambilla E, Donega M, Alfaro-Cervello C, Snider S, et al. Transplanted neural stem/precursor cells instruct phagocytes and reduce secondary tissue damage in the injured spinal cord. Brain. 2012;135(Pt 2):447–60.
Geim AK, Novoselov KS. The rise of graphene. Nat Mater. 2007;6(3):183–91.
Giboureau N, Som IM, Boucher-Arnold A, Guilloteau D, Kassiou M. Pet radioligands for the vesicular acetylcholine transporter (vacht). Curr Top Med Chem. 2010;10(15):1569–83.
Hedegaard C, Kjaer-Sorensen K, Madsen LB, Henriksen C, Momeni J, Bendixen C, et al. Porcine synapsin 1: syn1 gene analysis and functional characterization of the promoter. FEBS Open Bio. 2013;3:411–20.
Hu Y, Qu ZY, Cao SY, Li Q, Ma L, Krencik R, et al. Directed differentiation of basal forebrain cholinergic neurons from human pluripotent stem cells. J Neurosci Methods. 2016b;266:42–9.
Jiang Z, Song Q, Tang M, Yang L, Cheng Y, Zhang M, et al. Enhanced migration of neural stem cells by microglia grown on a three-dimensional graphene scaffold. ACS Appl Mater Inter. 2016;8(38):25069–77.
Ku SH, Lee M, Park CB. Carbon-based nanomaterials for tissue engineering. Adv Healthc Mater. 2013;2:244–60.
Lalwani G, Gopalan A, D’Agati M, Sankaran JS, Judex S, Qin YX, et al. Porous three-dimensional carbon nanotube scaffolds for tissue engineering. J Biomed Mater Res A. 2015;103(10):3212–25.
Langer R, Vacanti JP. Tissue engineering. Science. 1993;260(5110):920–6.
Li N, Zhang Q, Gao S, Song Q, Huang R, Wang L, et al. Three-dimensional graphene foam as a biocompatible and conductive scaffold for neural stem cells. Sci Rep. 2013;3:1604.
Loeblein M, Perry G, Tsang SH, Xiao W, Collard D, Coquet P, et al. Three-dimensional graphene: a biocompatible and biodegradable scaffold with enhanced oxygenation. Adv Healthc Mater. 2016;5(10):1177–91.
Mason C, Dunnill P. A brief definition of regenerative medicine. Regen Med. 2008;3:1–5.
Mufson EJ, Ginsberg SD, Ikonomovic MD, DeKosky ST. Human cholinergic basal forebrain: chemoanatomy and neurologic dysfunction. J Chem Neuroanat. 2003;26(4):233–42.
Nayak TR, Andersen H, Makam VS, Khaw C, Bae S, Xu X, et al. Graphene for controlled and accelerated osteogenic differentiation of human mesenchymal stem cells. ACS Nano. 2011;5(6):4670–8.
Shadjou N, Hasanzadeh M. Graphene and its nanostructure derivatives for use in bone tissue engineering: recent advances. J Biomed Mater Res A. 2016;104(5):1250–75.
Sudhof TC, Czernik AJ, Kao HT, Takei K, Johnston PA, Horiuchi A, et al. Synapsins: mosaics of shared and individual domains in a family of synaptic vesicle phosphoproteins. Science. 1989;245(4925):1474–80.
Takeichi M. The cadherins: cell-cell adhesion molecules controlling animal morphogenesis. Development. 1988;102(4):639–55.
Tozeren A, Wu S, Hoxter B, Xu W, Adamson ED, Byers SW. Vinculin and cell-cell adhesion. Cell Adhes Commun. 1998;5(1):49–59.
Yue C, Jing N. The promise of stem cells in the therapy of alzheimer’s disease. Transl Neurodegener. 2015;4:8.
You do not currently have access to this content.