Abstract

Goat milk-derived extracellular vesicles (GMEVs) are natural nanostructures with potential roles in nutrition and therapeutic delivery, yet their gastrointestinal fate remains poorly understood. Here, we systematically investigated the stability, microbial utilization, and biodistribution of GMEVs. Isolated GMEVs exhibited characteristic size distribution and bilayer morphology, with lipids dominated by glycerolipids (36.0%) and phospholipids (35.9%). They remained structurally stable in simulated digestion fluids but aggregated under colonic conditions. Fermentation assays showed GMEVs promoted microbial metabolism, increasing short-chain fatty acids and enriching Lactobacillus . Cellular uptake in Caco-2 cells occurred via caveolin- and clathrin-mediated pathways. In vivo fluorescence imaging in mice confirmed sequential passage through the gastrointestinal tract, with absorption by colonic enterocytes and concurrent microbial utilization, followed by detectable distribution to the liver and brain. Collectively, these findings establish GMEVs as resilient, multifunctional vesicles that couple nanocarrier capacity with prebiotic-like activity, highlighting their translational potential for oral drug delivery and microbiota-targeted interventions.