Composite nanofibrous membranes hold significant potential for the development of innovative biomedical materials and applications as their properties can be tuned by combining the synergistic effects of the nanofiller and the polymer matrix. Moreover, the nanofibrous structure can further enhance this feature thanks to the extremely high functional surface area. In the present work, a simple and fast method for the preparation of electrospun composite membranes based on cross-linked poly(ethylene oxide) (PEO) nanofibers and silver nanoparticles (Ag NPs) is proposed. For this purpose, a precursor of the Ag NPs (i.e., AgNO 3 ) is introduced into the aqueous solution of PEO in the presence of a suitable photoinitiator and a photo-cross-linker. Electrospinning of the solution is carried out, and the subsequent irradiation of the electrospun mat allows the simultaneous photoinduced cross-linking of PEO and the photoinduced synthesis of Ag NPs. The in situ generation of Ag NPs is confirmed through different characterization techniques (i.e., UV−vis spectroscopy, dynamic light scattering, and scanning electron microscopy). The structural, morphological, and mechanical properties of the resulting electrospun composite nanofibers are analyzed, and the effect of the content of the Ag NP precursor is evaluated. It is demonstrated that PEO fibers are chemically resistant and mechanically robust thanks to cross-linking, while Ag NPs induce significant antibacterial activity to the electrospun materials, which are also shown to be noncytotoxic by using both human fibroblasts and 3D-reconstructed skin equivalents. The proposed PEO-based nanofibrous mats decorated with Ag NPs thus exhibit promising potential as biomedical materials, especially for wound healing applications.