Engineered bioactive methylcellulose/gelatin composite hydrogel scaffolds with tuned micromechanics and nanotopography for enhanced myogenic differentiation of porcine muscle stem cells.
The escalating global demand for sustainable protein sources has accelerated research into cultivated meat. A critical requirement for large-scale production is developing edible, three-dimensional (3D) scaffolds that support cell growth and muscular tissue formation. This study developed and evaluated a biocompatible, cost-effective 3D scaffold for cultivated meat applications. Here, a novel methylcellulose/gelatin composite scaffold (4MC2G) was engineered via freeze-drying, demonstrating significant improvements over pure methylcellulose for myogenesis. Gelatin (G) incorporation significantly enhanced the physicochemical and biological performance of the scaffold. Mechanically, the 4MC2G scaffold exhibited a substantial increase in elastic modulus and displayed desirable nonlinear hyperelastic behavior. Microstructural analysis using SEM revealed that G incorporation refined the pore architecture (43.59 ± 14.21 μm) and created a more heterogeneous surface morphology. The cumulative release profile showed an initial burst release of G (~60.91 ± 0.81%) during the first 6 h, followed by a stabilized release rate. AFM confirmed that G incorporation enhanced the nanotopography and stiffness of both dry and hydrated scaffolds, which are critical for mechanotransduction. Additionally, the 4MC2G scaffold significantly promoted C2C12 myoblast adhesion, proliferation, and differentiation compared to its pure MC counterpart. Robust myogenic differentiation was confirmed by myotube formation, the upregulation of myosin heavy chain (MHC) and myogenin (MYOG), and the downregulation of paired box 7 (PAX7) mRNA expression. Finally, porcine muscle stem cell (PMSC) validation confirmed attachment, myotube formation, and MHC expression, establishing 4MC2G as a promising biocompatible and edible platform. These findings provide a valuable strategy for designing macromolecular composites for sustainable food production.