Hybrid 3D-printed/electrospun scaffolds drive myogenic differentiation of mesenchymal stem cells (MSCs).

The development of functional scaffolds that support cell differentiation and tissue regeneration remains a major challenge in regenerative medicine. In this study, we designed and fabricated hybrid scaffolds (HS) composed of poly(lactic-co-caprolactone) (PLA-PCL) by integrating 3D extrusion-based printing and electrospinning techniques. Two distinct 3D-printed architectures were explored-aligned filaments and grid patterns-which were subsequently coated with a layer of electrospun nanofibers to better mimic the anisotropic and hierarchical structure of native skeletal muscle tissue. Bone marrow porcine mesenchymal stem cells (p-MSCs) were seeded onto the scaffolds and cultured under myogenic conditions. To evaluate the progression of myogenic differentiation, we assessed the expression of early and late myogenic markers, MyoD and Myogenin respectively, at 14, 21 and 28 days. A comprehensive physical-mechanical characterization was performed, including morphological analysis, porosity measurements, and uniaxial tensile testing. The results demonstrated that the HS provided a biomimetic microenvironment that supported p-MSC attachment, viability, and differentiation. Notably, the HS aligned architecture enhanced the expression of myogenic markers compared to the grid design, suggesting a role of topographical cues in directing lineage commitment. These findings highlight the potential of dual-fabricated PLA-PCL scaffolds as a promising platform for guiding myogenic differentiation and may serve as a foundation for promoting functional skeletal muscle regeneration.