Guiding cellular behavior with micro/nanostructured cellulose acetate substrates fabricated using femtosecond laser lithography

The lack of efficient, reliable, and sustainable substrates for directed cell growth and tissue engineering remains a critical challenge in modern biomedical applications. Conventional cell culture platforms, such as flat and featureless surfaces, fail to replicate the complex microenvironments necessary for guiding cellular behavior, including contact guidance. This limitation is particularly significant in applications such as skeletal muscle tissue engineering, where the alignment and morphology of cells play a critical role in functionality. Additionally, the growing demand for eco-friendly materials necessitates the development of renewable and biocompatible alternatives to synthetic polymers. To address these challenges, we have fabricated micro- and nanopatterned substrates on cellulose acetate, a sustainable and biodegradable biopolymer, using a femtosecond laser. Ultrafast laser processing enables precise patterning of cellulose surfaces to achieve isotropic and anisotropic features of physiological relevance. Substrates with anisotropic grooves were evaluated for their ability to support and guide the alignment of mouse myoblast cells (C2C12), with the results demonstrating the potential of these engineered patterned substrates in supporting cell attachment, proliferation, and alignment. This work not only shows the potential of using cellulose acetate as an advanced cell culture platform, but also highlights femtosecond laser processing as a versatile tool for creating substrates that mimic natural extracellular environments. By addressing the key limitations of current cell culture methods, this study advances the development of sustainable, functional scaffolds for tissue engineering and regenerative medicine applications.