Lyophilized mesenchymal stromal cell-derived extracellular vesicles for the oral treatment of inflammatory bowel disease: a novel cell-free therapeutic strategy

BACKGROUND: Inflammatory bowel disease (IBD), encompassing ulcerative colitis (UC) and Crohn's disease (CD), is a chronic and relapsing inflammatory disorder of the gastrointestinal tract. Current standard treatments, including aminosalicylates, corticosteroids, immunomodulators, and biologic therapies, can induce and maintain remission; however, a considerable proportion of patients show limited responsiveness or experience adverse effects. This highlights the urgent need for safer and more effective therapeutic alternatives. Mesenchymal stromal cell-derived extracellular vesicles (MSC-EVs) represent a promising cell-free therapeutic modality owing to their immunomodulatory and reparative capabilities. These vesicles replicate many of the therapeutic benefits of mesenchymal stromal cells (MSCs) while avoiding the risks associated with conventional cell-based treatments. Nevertheless, clinical translation of MSC-EVs for IBD management remains constrained by issues such as limited storage stability and invasive delivery routes. METHODS: Extracellular vesicles (EVs) were isolated from the culture supernatant of human umbilical cord-derived MSCs via tangential flow filtration (TFF) followed by ultracentrifugation, and were characterized using transmission electron microscopy, western blotting, and nanoflow cytometry. To enhance their stability, MSC-EVs were lyophilized with trehalose and mannitol serving as cryoprotectants. The lyophilized powder was subsequently encapsulated in enteric-coated capsules for oral administration. The therapeutic efficacy of this formulation was evaluated in a dextran sulfate sodium (DSS)-induced rat colitis model. Disease activity was recorded daily, and post-experimental analyses included histopathology, immunohistochemistry, quantitative real-time polymerase chain reaction (qRT-PCR), and serum biochemistry to evaluate hepatorenal function. RESULTS: A robust lyophilization protocol was successfully established to enhance MSC-EV stability, along with an innovative oral delivery system for IBD therapy. In DSS-induced colitis rats, lyophilized MSC-EVs markedly alleviated disease severity, as indicated by a reduced disease activity index (DAI), preserved colon length, and improved histological architecture. MSC-EV administration restored intestinal barrier integrity, modulated cytokine expression profiles, mitigated systemic inflammation, and improved hepatic and renal function. CONCLUSIONS: This study demonstrated the therapeutic efficacy of a lyophilized, orally administered MSC-EV formulation in experimental colitis. This approach not only enhances vesicle stability and simplifies storage but also provides a non-invasive, patient-friendly route of administration. These findings underscore the strong translational potential of lyophilized MSC-EVs as a practical and effective cell-free therapeutic strategy for IBD.