Ana I. Salazar-Puerta, Sara Kheirkhah, Jonathan Stranan, Hallie Harris, Nada Khattab, Megumi Fukuda, Grant Barringer, Carlos A Vasquez-Martinez, Samuel Cortes, Neil Ott, William Lawrence, Devleena Das, Jordan Moore, Tiam M. Saffari, Juan D. Salazar-Gil, Diego Alzate-Correa, Junyan Yu, Kavya Dathathreya, Natalia Higuita‐Castro, Tatiana Cuellar-Gaviria, W. David Arnold, Amy M. Moore, Daniel Gallego Pérez
Peripheral nerve injuries (PNIs) remain a major clinical challenge, often leaving patients with lifelong sensory, motor, and functional impairments despite surgical repair. While gene and cell therapies hold promise, their translation has been hampered by the lack of safe, efficient, and targeted delivery strategies. Here, we introduce vasculogenic tissue nanotransfection (TNT) as a nonviral, reprogramming-based therapeutic platform to enhance nerve regeneration to augment surgical reconstruction. This one-time, millisecond-scale intervention reprograms resident cells in situ toward a vasculogenic phenotype, fostering neovascularization and vascular remodeling to support axonal regeneration. Through integrated in vitro screening and in vivo validation, we identified an optimized formulation of vasculogenic genes ( Etv2 , Fli1 , and Foxc2 ; EFF ) that maximized reprogramming efficiency and regenerative potential. In a long-segment nerve defect model reconstructed with isografts, TNT-mediated delivery of EFF markedly improved functional recovery, including grip strength and muscle contractility, accompanied by increased vascular density and myelinated axon counts. Together, these findings establish TNT-mediated vasculogenic reprogramming as a transformative adjunct to surgical repair of PNIs, offering a clinically translatable strategy to accelerate nerve regeneration and restore function.