Exploiting topography and stiffness to promote muscle fibre self-organization in vitro

During tissuegenesis and organogenesis processes cell behavior is profoundly affected by biophysical and biochemical signals derived from surrounding extracellular matrix (ECM). Cell fate is influenced by material properties and more specifically cell-material crosstalk plays a crucial role in the architecture and organization of tissues. For instance, microstructure and stiffness of ECM are known to control and guide specific morphogenetic events. In many cases, the morphological structure of tissues, such as muscle, is closely linked to their function. Emerging evidence suggests that the self-assembly and organization of muscle require localized geometrical and biochemical cues that guide myofibrillogenesis. Here we report the development of nanoengineered platforms for in vitro skeletal muscle self-assembly. The aim of this study is to define the optimal set of initial conditions that direct and control muscle supracellular self-organization. Our findings emphasize the pivotal role of substrate nanotopography, stiffness and adhesivity in guiding cell orientation and fusion, key factors for muscle tissue engineering. These insights offer a platform for engineering functional muscle tissues with native-like structural and functional properties. This work provides valuable information for the optimization of substrate properties to regulate cellular self-organization and tissue morphogenesis in vitro, with potential applications in regenerative medicine and tissue engineering.