Tunable anisotropic nano/micro-topography aligns myotubes and accelerates maturation

Skeletal muscle regeneration requires biomaterials that replicate the anisotropic architecture of native tissue. However, existing micro/nanofabrication approaches for creating aligned topographies-such as photolithography and nanoimprinting-typically require photomasks, master molds, or cleanroom facilities, which increase cost and complexity and limit their preclinical and clinical translation. Here, we present a simple, mask- and mold-free bottom-up strategy to fabricate one-dimensional (1D, anisotropic) and two-dimensional (2D, isotropic) wrinkled MXene multilayer coatings with tunable nano- to microscale topographies. MXene multilayer films were assembled via layer-by-layer (LbL) deposition and thermally shrunk to generate controlled wrinkle geometries. C2C12 myoblasts cultured on micro-wrinkled MXene surfaces exhibited markedly enhanced alignment, elongation, and proliferation compared to planar and nano-textured substrates. Gene expression analysis revealed strong upregulation of key myogenic markers (MyoD, Myogenin, Acta1, Myh3, and Mck) that was largely sustained over 7 days of differentiation, accompanied by a higher fusion index and the formation of elongated, multinucleated myotubes with organized α-actinin striations. These findings demonstrate that the anisotropic topography of MXene coatings effectively promotes myogenic differentiation and structural maturation, establishing a tunable and accessible platform for aligned muscle tissue engineering.