Functionalized Multi‐Walled Carbon Nanotube Enhanced Myogenic Differentiation for Aligned Topography‐Induced Skeletal Muscle Engineering

Abstract Skeletal muscle engineering utilizing bio‐activators and myogenic cells to regenerate tissues for volumetric muscle loss offers a promising alternative to tissue grafts. Modified biointerfaces with aligned micro‐scale topography and electroconductivity are critical for directing cellular behavior toward functional muscle constructs. This study modified polydimethylsiloxane (PDMS) with aligned surface topography and functionalized multi‐walled carbon nanotubes (fCNTs), creating a conductive scaffold (0.11 µScm ⁻¹ vs original 0.51 nScm ⁻¹ ) with regulated hydrophilicity (76 ± 2° vs original 50 ± 10° in water contact angle) and enhanced protein absorption. The fCNT‐wrinkled surfaces maintained >90% cell viability while promoting aligned myotube formation. Specifically, fCNT integration with aligned topography increased myotube length from 303.74 ± 27.61 µm to 441.63 ± 10.27 µm and elevated fusion index to 40.43% ± 2.67% within three differentiation days. Immunostaining confirmed enhanced myogenic maturation through improved cell alignment and nuclei organization. These biophysical modifications synergistically accelerated myoblast differentiation while maintaining cytocompatibility by combining electrical conductivity, optimized wettability, and directional cues. The demonstrated capacity to physiologically mimic native muscle microenvironments highlights this strategy's potential for improving muscle regeneration therapies through precise control of surface‐electrotopographical properties.