Fig. 4.

3D MnO₂ hybrid nanoscaffolds self-assembled with ECM proteins. a, b Schematic diagram illustrating the mechanism for the self-assembly of ECM protein (laminin) and MnO₂ nanosheets through a non-covalent crosslinking mechanism. This mechanism was utilized to synthesize MnO₂ laminin hybrid nanoscaffolds, and iPSC-NSCs cultured on MnO₂-laminin hybrid nanoscaffolds successfully differentiated into mature (MAP2+) neurons after 6 days (b). c–e Representative SEM images of MnO₂ laminin hybrid nanoscaffolds (c, d), and iPSC-NSC-encapsulated hybrid nanoscaffolds (e). f–h Immunostaining results on neuronal markers (TuJ1, labeled with green) demonstrate significant enhancements of cell adhesion and neuronal differentiation of iPSC-NSCs differentiated on MnO₂ laminin hybrid nanoscaffolds (h) compared to both control (glass, f) substrates and MnO₂ hybrid nanoscaffolds (g). Average percentage of neuron and neurite lengths were labeled on the bottom left of each image. i Time-lapse calcium imaging results of iPSC-NSC differentiated neurons on MnO₂ laminin hybrid nanoscaffolds. j, k representative immunostaining images on mature neuronal markers (i: MAP2 and k: Synapsin 1, labeled with green) from iPSC-NSC-differentiated neurons on MnO₂ laminin hybrid nanoscaffolds. Scale bars: c 10 μm; d 500 nm; e–h 100 μm; j, k 100 μm