Speaker
Description
The regeneration of skeletal muscle (SM) tissue and its interfaces—namely the myotendinous junction (MTJ) and neuromuscular junction (NMJ)—remains a significant challenge in both engineering and clinical domains. Recent advances in biofabrication have begun to address these hurdles, enabling the creation of biomimetic architectures with precise spatial control over cellular organization and mechanical properties.
Here, we present a comprehensive investigation into the use of microfluidics-assisted 3D rotary wet-spinning (RoWS) for the scalable fabrication of functional, anisotropic, and vascularized hydrogel-based constructs tailored for skeletal muscle, MTJ, and microvascular tissue engineering. Our recently developed high-throughput RoWS platform enables the continuous production of core–shell hydrogel microfibers, which serve as modular building blocks for hierarchically structured 3D scaffolds. By optimizing key parameters—including flow rates, rotational speed, and fiber composition—we produced highly aligned fiber bundles that guide uniaxial cell alignment and promote myogenic differentiation.
Using this platform, we demonstrated that constructs encapsulating human pericytes exhibit significantly enhanced myogenic maturation compared to traditional 2D cultures and 3D bulk hydrogels. Proteomic analysis revealed a distinct molecular signature associated with the anisotropic 3D microenvironment, marked by upregulation of contractile and extracellular matrix proteins. In vivo studies in a murine model of volumetric muscle loss further validated the regenerative potential of these constructs, which showed successful engraftment and restoration of muscle architecture.
Building on these findings, we adapted the RoWS system to engineer MTJ-like hydrogel yarns through sequential wet-spinning of C2C12 myoblasts and NIH 3T3 fibroblasts. This strategy recapitulates the graded biological and structural organization of the native muscle–tendon interface. The resulting constructs displayed uniaxial cellular alignment and expression of tissue-specific markers (e.g., MyHC, collagen I/III), and featured hallmark finger-like projections at the muscle-tendon transition zone—mimicking the native MTJ microarchitecture with high fidelity.
To address the crucial need for vascularization in engineered muscle constructs, we also developed a co-culture system comprising human mesenchymal stem cells (hMSCs) and human umbilical vein endothelial cells (HUVECs) within a fibrin-based core. The RoWS facilitated the formation of aligned, capillary-like networks and robust expression of endothelial markers such as CD31, confirming successful microvascularization.
Currently, we are extending our platform to incorporate neuro-mesodermal progenitors (NMPs) for the fabrication of “tubuloids” that may emulate the structural and functional complexity of the NMJ.
Collectively, these findings underscore the versatility and translational promise of the RoWS biofabrication platform for engineering anisotropic, multicellular, and vascularized musculoskeletal tissue constructs. With its high functional performance, biomimetic precision, and scalability, this platform offers a compelling foundation for both next-generation in vitro models and future therapeutic strategies in tissue regeneration.
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