Human Skeletal Muscle\nMyoblast Culture in Aligned\nBacterial Nanocellulose and Commercial Matrices

Bacterial nanocellulose (BNC) is a durable, flexible,\nand dynamic\nbiomaterial capable of serving a wide variety of fields, sectors,\nand applications within biotechnology, healthcare, electronics, agriculture,\nfashion, and others. BNC is produced spontaneously in carbohydrate-rich\nbacterial culture media, forming a cellulosic pellicle via a nanonetwork\nof fibrils extruded from certain genera. Herein, we demonstrate engineering\nBNC-based scaffolds with tunable physical and mechanical properties\nthrough postprocessing. Human skeletal muscle myoblasts (HSMMs) were\ncultured on these scaffolds, and in vitro electrical stimulation was\napplied to promote cellular function for tissue engineering applications.\nWe compared physiologic maturation markers of human skeletal muscle\nmyoblast development using a 2.5-dimensional culture paradigm in fabricated\nBNC scaffolds, compared to two-dimensional (2D) controls. We demonstrate\nthat the culture of human skeletal muscle myoblasts on BNC scaffolds\ndeveloped under electrical stimulation produced highly aligned, physiologic\nmorphology of human skeletal muscle myofibers compared to unstimulated\nBNC and standard 2D culture. Furthermore, we compared an array of\nmetrics to assess the BNC scaffold in a rigorous head-to-head study\nwith commercially available, clinically approved matrices, Kerecis\nOmega3 Wound Matrix (Marigen) and Phoenix as well as a gelatin methacryloyl\n(GelMA) hydrogel. The BNC scaffold outcompeted industry standard matrices\nas well as a 20% GelMA hydrogel in durability and sustained the support\nof human skeletal muscle myoblasts in vitro. This work offers a robust\ndemonstration of BNC scaffold cytocompatibility with human skeletal\nmuscle cells and sets the basis for future work in healthcare, bioengineering,\nand medical implant technological development.