A green fabrication process (organic solvent-free) of artificial scaffolds is required in tissue engineering field. In this work, a series of aligned three-dimensional (3D) scaffolds are made from biodegradable waterborne polyurethane (PU) emulsion via directional freeze–drying method to ensure no organic byproducts. After optimizing the concentration of polymer in the emulsion and investigating different freezing temperatures, an aligned PUs scaffold (PU14) generated from 14?wt% polymer content and processed at ?196°C was selected based on the desired oriented porous structure (pore size of 32.5?±?9.3?μm, porosity of 92%) and balanced mechanical properties both in the horizontal direction (strength of 41.3?kPa, modulus of 72.3?kPa) and in the vertical direction (strength of 45.5?kPa, modulus of 139.3?kPa). The response of L929 cells and the regeneration of muscle tissue demonstrated that such pure material-based aligned 3D scaffold can facilitate the development of orientated cells and anisotropic tissue regeneration both in vitro and in vivo. Thus, these pure material-based scaffolds with ordered architecture have great potentials in tissue engineering for biological anisotropic tissue regeneration, such as muscle, nerve, spinal cord and so on.
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