Abstract Background Inflammatory bowel disease (IBD) is a chronic, relapsing inflammation disorder of the gastrointestinal tract characterized by disrupted intestinal epithelial barrier function. Despite advances in treatment, including biologic agents, achieving sustained remission remains challenging for many IBD patients. This highlights the urgent need for novel therapeutic strategies. Milk-derived extracellular vesicles (MDEs) have emerged as a promising therapeutic option. Methods MDEs were isolated from raw cow milk through sequential centrifugation, filtration, and ultracentrifugation. Their morphology and size were characterized by transmission electron microscopy, dynamic light scattering, and Western blot analysis of CD81. Functional studies were performed using intestinal epithelial cell lines, a murine model of dextran sulfate sodium (DSS)-induced colitis, and patient-derived colon organoids (PDCOs) from IBD and non-IBD tissues. RNA was extracted from MDEs and cells for mRNA and miRNA expression profiling by qRT-PCR. MDE uptake was visualized by fluorescence microscopy, and proliferation and viability were assessed by Ki67 immunostaining and CCK-8 assays. Results Using a murine model of DSS-induced colitis, we observed that MDEs significantly ameliorated disease symptoms. The upregulation of β-catenin, a crucial mediator of Wnt signaling, in colonic tissues, suggests that MDEs may facilitate epithelial regeneration and restore barrier function. In PDCOs, MDEs were internalized and modulated the expression of key signaling molecules, such as the upregulation of β-catenin, cyclin D1, and the proliferation marker Ki67, indicating their potential to promote IEC proliferation and intestinal barrier repair. Importantly, MDEs demonstrated selective activity by downregulating β-catenin and cyclin D1 in colon cancer cells, leading to reduced proliferation. Conclusion Our findings demonstrate that MDEs promote intestinal epithelial recovery while maintaining selective anti-proliferative activity in malignant cells. These results highlight MDEs as a safe, naturally derived therapeutic candidate for intestinal barrier repair in IBD and related inflammatory disorders. Conflict of interest: Reif, Shimon: CMO of EXOSOMM Birimberg Schwartz, Liron: No conflict of interest Grunewald, Myriam: No conflict of interest Duran, Deborah: No conflict of interest Sebbag-Sznajder, Naama: No conflict of interest Musseri, Mirit: No conflict of interest Toledano, Tirtsa: No conflict of interest Dr. Golan-Gerstl, Regina: CTO of EXOSOMM.
Reif et al. (Thu,) studied this question.