Osmotic Remodeling of Extracellular Vesicles for Precision Nanomedicine
Francesca Susa, Lorenzo Arnaboldi, Veronica De Giorgis, Marco Ghirimoldi, Elettra Barberis, Marco Brucale, Francesco Valle, Mario Malerba, Marta Vallino, Angelo Musicò, Roberto Frigerio, Alessandro Gori, Vera Mugoni, Claudia Voena, Roberto Pisano, Fabrizio Pirri, Silvia Arpicco, Marcello Manfredi, Tania LimongiABSTRACT
Extracellular vesicles (EVs) are increasingly explored as diagnostic and therapeutic platforms. Translation is constrained by protocols that must balance efficacy and safety, especially for EVs from tumor or immortalized cells that may carry oncogenic or immunostimulatory cargo. This study presents a rapid and reproducible engineering of EVs using bidistilled water as the sole reagent. The hypotonic environment induces osmotic lysis of EVs isolated from B lymphocytes, generating a remodeled vesicle population. The process markedly reduces a broad spectrum of intraluminal components with potential immunogenic or tumor‐promoting activity, while preserving the vesicular membrane scaffold. Osmotic remodeling is associated with reorganization of membrane‐associated lipids and proteins and with a preferential uptake of the remodeled vesicles by tumor cells over non‐malignant counterparts in our in vitro model. The engineered EVs retain structural integrity and are amenable, at a proof‐of‐concept level, to reloading with defined bioactive compounds, suggesting their potential as customizable delivery systems. Overall, this osmotic remodeling strategy provides a versatile technological platform for the generation of membrane‐derived vesicles with tunable properties and promising applicability in drug delivery, nanodiagnostics, and immune modulation, whose translational performance and safety will need to be established in source‐ and indication‐specific preclinical studies.