Mannose-cRGD Engineered Lactobacillus helveticus-Derived Exosomes Enhance Loaded Pterostilbene Bioavailability: From Dual Insights of Membrane Lipid Orderliness and Dual-Targeting Endocytosis Modulation.

Exosomes derived from probiotics are emerging as innovative oral nanocarriers but face challenges in targeted delivery and stability. Here, Lactobacillus helveticus-derived exosomes (LHexo) were dual-functionalized with mannose and cRGD (McRLHexo), yielding improved membrane lipid ordering, denser PEG brush layer, and distinct "cloud-like" surface nanostructure. McRLHexo exhibited enhanced stability under UV (transmittance reduction > 98%), thermal, and digestive conditions (drug retention >86%), and increased pterostilbene (PTS) loading. Functionally, McRLHexo@PTS achieved efficient epithelial translocation via cRGD-integrin pathways and macrophage targeting via Man/cRGD dual-recognition. In DSS-induced colitis mice, McRLHexo showed optimal targeting accumulation and uptake in the colon. Importantly, McRLHexo@PTS markedly alleviated colitis, restoring body weight, colon length, and epithelial integrity while reducing inflammatory cytokines and oxidative stress and enhancing antioxidant defenses. This study establishes a dual-targeted probiotic exosome platform for oral therapy and provides insights into ligand-membrane engineering for precision intestinal delivery.