Combination therapy using intestinal organoids and their extracellular vesicles for inflammatory bowel disease complicated with osteoporosis.

Background: Inflammatory bowel disease (IBD) with osteoporosis (OP) exhibits a clinically significant comorbidity, for which no effective treatment is currently available. Intestinal organoids (IOs), engineered through three-dimensional (3D) coculture systems, demonstrated intrinsic regenerative potentials. Additionally, extracellular vesicles derived from IOs (IOEVs) have been identified as potent nanoscale mediators capable of modulating intestinal inflammation. Methods: In this study, we successfully established IOs and isolated IOEVs. miRNA sequencing in IOEVs revealed IBD-associated miRNAs, which may alleviate inflammatory response and have osteogenic effects. An in vitro model of IBD was established using lipopolysaccharide (LPS) to induce inflammation. Additionally, the dextran sulfate sodium (DSS)-induced IBD mouse model was employed to evaluate in vivo effects. Results: In the LPS-induced in vitro model, treatment with IOs and IOEVs resulted in reduced cell necrosis and apoptosis. In DSS-induced IBD mouse models, treatment led to restoration of body weight and colon morphology. Histological assessment revealed an increase in intestinal crypts and normalization of tissue architecture. Immunological analyses showed upregulation of ZO-1 and Ki67 and downregulation of Caspase-3, suggesting enhanced mucosal barrier integrity and cellular proliferation with decreased apoptosis. Cytokine profiling showed downregulation of pro-inflammatory cytokines TNF-α, IL-1β, IL-6 and upregulation of anti-inflammatory cytokine IL-10. Importantly, the combination of IOs and IOEVs reversed osteoporosis progression in IBD, improving bone mass and quality. Conclusion: Collectively, these multimodal findings establish a novel paradigm for gut-bone axis modulation through organoid-derived biologics, offering a promising therapeutic strategy for managing IBD-associated osteoporosis. The translational potential of this article: This study highlights the translational potential of intestinal organoids and their extracellular vesicles as a dual-action biologic therapy that alleviates intestinal inflammation and reverses bone loss in IBD-associated osteoporosis. The identification of functional miRNAs within IOEVs supports their development as minimally invasive, cell-free therapeutics for systemic complications in inflammatory disease.