3D anisotropic conductive fibers electrically stimulated myogenesis

Recapitulation of in vivo environments that drive muscle cells to organize into a physiologically relevant 3D architecture remains a major challenge for muscle tissue engineering. To recreate electrophysiology of muscle tissues, electroactive biomaterials have been used to stimulate muscle cells with exogenous electrical fields. In particular, the use of electroactive biomaterials with an anisotropic micro-/nanostructure that closely mimic the native skeletal-muscle extracellular matrix (ECM) is desirable for skeletal muscle tissue engineering. Herein, we present a hierarchically organized, anisotropic, and conductive Polycaprolactone/gold (PCL/Au) scaffold for guiding myoblasts alignment and promoting the elongation and maturation of myotubes under electrical stim-ulation. Culturing with H9c2 myoblasts cells indicated that the nanotopographic cues was crucial for nuclei alignment, while the presence of microscale grooves effectively enhanced both the formation and elongation of myotubes. The anisotropic structure also leads to anisotropic conductivity. Under electrical stimulation, the elongation and maturation of myotubes were significantly enhanced along the anisotropic scaffold. Specifically, compared to the unstimulated group (0 V), the myotube area percentage increased by 1.4, 1.9 and 2.4 times in the 1 V, 2 V, 3 V groups, respectively. In addition, the myotube average length in the 1 V group increased by 1.3 times compared to that of the unstimulated group, and significantly increased by 1.8 and 2.0 times in the 2 V, 3 V groups, respectively. Impressively, the longest myotubes reached more than 4 mm in both 2 V and 3 V groups. Overall, our conductive, anisotropic 3D nano/microfibrous scaffolds with the application of electrical stimulation provides a desirable platform for skeletal muscle tissue engineering.