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Electro-assembled dynamically adaptive collagen can recapitulate the biology’s ability to tune structures and functions. 

Abstract Janus porous biomaterials are gaining increasing attention and there are considerable efforts to develop simple, rapid, and scalable methods capable of tuning micro‐ and macro‐structures. Here, a single‐step electro‐fabrication method to create a Janus porous film by the electrodeposition of the amino‐polysaccharide chitosan is reported. Specifically, a Janus structure emerges spontaneously when electrodeposition is performed at sub‐ambient temperature (0–5 °C). Sub‐ambient temperature electrodeposition experiments show that: a Janus microstructure emerges (potentially as the result of a subtle alteration of the intermolecular interactions responsible for self‐assembly); important microstructural features (pore size, porosity, and thicknesses) can be tuned by conditions; and this method is readily scalable (vs serial printing) and can yield complex tubular structures with Janus faces. In vitro studies demonstrate anisotropic cell guidance, and in vivo studies using a rat calvarial defect model further confirm the beneficial features of such Janus porous film for guided bone regeneration. In summary, these results further demonstrate that electro‐fabrication provides a simple and scalable platform technology for the controlled functional structures of soft matter for applications in regenerative medicine.