Protocol for fabrication of nanosubstrate embedded with nanogroove topography coated by a layer of nanocomposite for neuronal differentiation.

Neurodegenerative diseases mainly affect the vital characteristics of neurons, including axons. The fabricated nanotopographies can cause axonal regeneration manipulating the migration and differentiation of cells. Here, we present a protocol for the fabrication of nanosubstrate incorporated with nanogroove topography with a coated layer of polyaniline-chitosan (PANI-C) nanocomposite. We describe steps for investigating differences between bulk polydimethylsiloxane (PDMS) sheets and embedding sheets with nanogrooves. We then detail procedures for coating with a PANI-C nanocomposite layer on the substrate. For complete details on the use and execution of this protocol, please refer to Afsharian et al..