Chronic diabetic glucolipotoxicity is associated with progressive skeletal muscle atrophy, bone loss, cellular senescence, and ferroptotic injury. Because ferroptosis inhibition alone may not restore compromised myogenic capacity, we developed a combined stem cell-conditioning strategy integrating fullerol priming with the stem cell screen-enrich-combine(-biomaterials) circulating system (SECCS). This integrated three-dimensional culture platform uses porous β-TCP granules and intermittent medium circulation to generate BMSCs-derived extracellular vesicles (EVs). Among 2D-EVs, F-2D-EVs, SECCS-EVs, and F-SECCS-EVs, F-SECCS-EVs exhibited the strongest anti-ferroptotic and pro-regenerative effects. In glucolipotoxicity-challenged myoblasts, F-SECCS-EVs attenuated ferroptotic injury and enhanced proliferation and myogenic differentiation. Transcriptomic and functional experiments identified FOXO3 as an important mediator linking antioxidant defense with myogenic programs. In diabetic mice, F-SECCS-EVs improved motor function, muscle strength, myofiber cross-sectional area, and myogenic marker expression while modulating macrophage polarization. Free fullerol, 2D-EVs, and F-2D-EVs produced limited recovery, whereas SECCS-EVs were less effective than F-SECCS-EVs. These muscle benefits were accompanied by favorable changes in tibial bone structure. Collectively, these findings support fullerol-primed SECCS culture as a strategy for coordinating ferroptosis control with myogenic recovery in diabetes-associated musculoskeletal degeneration.
