Tailorable Fiber Composite Cell Sheet For Wound Healing

While mesenchymal stem cell (MSC) sheets hold huge promise for tissue repair, their clinical translation is hindered by structural instability and compromised cell viability. To overcome these limitations, we developed tunable fibrous sheet composites that enable precise control over sheet maturation time and architecture, providing an engineered solution for standardized, homogeneous cell sheet fabrication. In vitro analyses assessed apoptosis/senescence markers (qPCR: Bax/Caspase3/p21; IF: γH2A.X/p53) and stemness properties (SOX2/Nanog IF; trilineage differentiation‌). Functional profiling of LPS-activated P-CS secretomes revealed epithelial migration (HaCaT scratch assay), angiogenesis (HUVEC tube formation assay), and macrophage polarization (BMDM CD206+/iNOS+ ratio). In vivo tracking in rat full-thickness skin wounds and oral mucosal defects demonstrated cellular retention dynamics and tissue regeneration capacity. In vitro, P-CSs exhibited reduced apoptosis and senescence markers while maintaining stemness. Under LPS stimulation, P-CSs retained proliferative capacity and suppressed apoptosis-related genes. Conditioned medium from LPS-treated P-CSs promoted migration, tubulogenesis, and M2 polarization. In vivo, P-CSs demonstrated dual regenerative-immunomodulatory efficacy: enhancing DFSCs retention and tissue regeneration via accelerated re-epithelialization/angiogenesis while suppressing pro-inflammatory Th17/M1 macrophage infiltration and promoting reparative M2 macrophage polarization. A tailorable ECM-mimetic nanofiber composite cell sheet (P-CSs) with enhanced structural adaptability, preserved bioactivity, immunomodulatory activity, and regenerative potential, providing a standardized platform for scalable clinical-grade production of functional grafts to accelerate translation.