Sarcopenia is an age-related skeletal muscle disorder characterized by the progressive loss of muscle mass and function, resulting in physical disability and mortality. Aberrant reactive oxygen species (ROS) generation and oxidative tissue damages in skeletal muscles have been implicated as one of key causative factors in the multiple mechanisms of sarcopenia. Tobacco smoke contains many toxic chemicals including highly unstable free radicals and ROS. Currently, heated tobacco products (HTPs) are widely used as substitutes for cigarette smoking around the world. However, it is poorly understood the effects of these new generation tobacco products on skeletal muscles. In the present study, we investigated the muscular injury induced by cigarette smoke extract (CSE) and HTPs using murine myoblast C2C12 cells as an in-vitro model. The exposure of either CSE or HTPs for 24 h decreased the cell viability in C2C12 myoblasts, which protected by treatment with 1 mM N-acetylcysteine (NAC). In addition, the differentiation to myotube and the expression of myogenic marker protein were suppressed by CSE or HTPs exposure. Interestingly, the formation of stress fibers was disrupted by the treatment with CSE, and the aberrant membrane protrusion was observed. These abnormalities in cytoskeletons and cell shapes were also rescued by the treatment of NAC. These findings suggested that CSE and/or HTPs can damage skeletal muscles through the impairment of cytoskeleton formation, which may contribute to the progress of muscular damages, and age-related muscular diseases.