Wound infections by Gram-negative bacteria are a significant challenge for medical trials, in part because of antibiotic resistance and robust immune evasion mechanisms. Such infections often cause extended hospitalizations, increased healthcare costs, and high morbidity and mortality rates. Biofilms and endotoxins from Gram-negative bacteria, contributing to chronic inflammation and delayed wound healing, further complicate their management. Most conventional therapeutic approaches, including broad-spectrum antibiotics and wound debridement, are often inadequate and thus represent a critical demand for novel and effective strategies. Eukaryotic extracellular vesicles (EVs), involving microvesicles and exosomes, have emerged as promising therapeutic tools because of their natural biocompatibility and multifunctional properties. EVs play a pivotal role in wound healing through mechanisms such as immune modulation, promotion of angiogenesis and tissue regeneration, antimicrobial action, and the activation of paracrine signaling pathways. EVs offer a holistic approach to infections and tissue repair by delivering bioactive molecules such as cytokines, growth factors, miRNAs, and antimicrobial peptides. Their potential to disrupt bacterial biofilms, neutralize endotoxins, and stimulate regenerative processes positions them as transformative agents in addressing Gram-negative bacterial infections. This mini-review, therefore, discusses the therapeutic mechanisms and clinical implications of eukaryotic EVs in wound healing, with their promise to revolutionize infection management by reducing dependence on antibiotics.