Oriented surfaces of adsorbed cellulose nanowhiskers promote skeletal muscle myogenesis

Cellulose nanowhiskers (CNWs) are high-aspect-ratio rod-like nanoparticles prepared via partial hydrolysis of cellulose. For the first time, CNWs have been extracted from the marine invertebrate Ascidiella aspersa, yielding animal-derived CNWs with particularly small diameters of only a few nanometres. Oriented surfaces of adsorbed CNWs were prepared using a flexible and facile spin-coating method, allowing the modulation of CNW adsorption and relative orientation. Due to the shape and nanoscale dimensions of the CNWs, C2C12 myoblasts adopted increasingly oriented morphologies in response to more densely adsorbed and oriented CNW surfaces. In addition, the degree of myoblast fusion was greatest on the highly oriented CNW surfaces, and even low-orientation CNW surfaces promoted more extensive fusion than flat control surfaces. Highly oriented multinuclear myotubes formed on the oriented CNW surfaces and fibrillar fibronectin deposited on the surfaces was also modelled in a highly oriented arrangement after only 4 days in culture. With a mean feature height of only 5-6 nm, the CNW surfaces present the smallest features ever reported to induce contact guidance in skeletal muscle myoblasts, highlighting the potential for nanoscale materials for engineering oriented tissues such as skeletal muscle.