Abstract
Surface micro- and nanostructures profoundly affect the functional performance of nerve regeneration implants by modulating neurite responses. However, few studies have investigated the impact of discrete nanostructures, such as nanopillars and nanoholes, and their combination with microgrooves on neurite outgrowth and alignment. Furthermore, numerous techniques have been developed for surface micro-/nanopatterning, but simple and low-cost approaches are quite limited. In this work, we show that nanopillars and nanoholes, and their combination with microgrooves, can be patterned on polyurethane (PU) films using a low-cost, reusable photoresist master mold prepared via nanosphere lens lithography and UV-LED photolithography, with specific "reinforcement" methods for overcoming the inherent drawbacks of using photoresist masters. We show that the PU nanopillars and nanoholes increase the neurite length of pheochromocytoma 12 (PC12) cells through unique growth cone interactions. Moreover, we demonstrate, for the first time, that hierarchically patterned nano-/microstructured PU films enhance both PC12 neurite elongation and alignment, showing the potential use of our proposed method for the micro-/nanopatterning of polymers for nerve tissue engineering.