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. 2018 Aug 8;9:3147. doi: 10.1038/s41467-018-05599-2

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — Enhanced stem cell differentiation using biodegradable MnO₂ hybrid nanoscaffolds. a Representative TEM (Transmission Electron Microscopy) of atomic-thin layered MnO₂ nanosheets. Inset: HR (High Resolution) TEM image (image size: 18 Å by 14 Å). b A schematic diagram describing the intermolecular binding between MnO₂ nanosheets and selected functional groups that are commonly existent in ECM proteins and biomolecules. c A significantly upregulated ECM protein (laminin) binding towards 2D MnO₂ nanosheet, compared to control substrate [etched glass and polycaprolactone (PCL) substrates). These upregulated laminin binding was studied by a BCA protein assay. Data are mean ± s.d. n = 3, **P < 0.01 by one-way ANOVA (Analysis of variance) with Tukey post-hoc test. d, e, By modeling small molecule-nanosheet interactions (d), we summarized differential binding affinities of MnO₂ nanosheets towards of a library of functional groups (e). These simulation results aligned well with experimentally observed enhanced laminin binding, as laminin structures are rich in both amino and aromatic moieties. Exemplary molecule in d: toluene. Carbon and hydrogen atoms are colored in black and gray, respectively. f A representative SEM image showing the layered structure and homogeneous surface of MnO₂ hybrid nanoscaffolds fabricated using the vacuum filtration technique. Inset photograph: a free-standing MnO₂ hybrid nanoscaffold fabricated by vacuum filtration and manipulated by a tweezer. g A schematic diagram illustrating the proposed mechanism for the enhanced neuronal differentiation on MnO₂ hybrid nanoscaffolds. Red color indicates GAP43; Orange indicates FAK; Green indicates integrin complex. h, i Representative immunostaining data supporting the significantly enhanced neuronal differentiation and neurite outgrowth of iPSC-NSCs on MnO₂ hybrid nanoscaffolds (i) compared to control substrates (h). Cell nuclei are in blue and TuJ1 is pseudocolored in green. Average percentages of neuron and neurite lengths were measured using NeuronJ software (h: n = 9; i: n = 12) and labeled on the top right of each image. Scale bars: a 25 nm; f 500 nm; h, i 100 μm