Immortalized Human Nasal Epithelial Cells-SV40

MSC-sEV Restored Nasal Barrier Function in Allergic Rhinitis Via Mir-143-GSK3B in Human Nasal Epithelial Cells

The nasal epithelial barrier is critical for defending against pathogens and allergens, with dysfunction implicated in allergic rhinitis (AR). Mesenchymal stem cell-derived small extracellular vesicles (MSC-sEV) show anti-inflammatory and regenerative potential, but their effects on nasal epithelial cells are unknown. Xu's team investigated MSC-sEV's therapeutic effects on AR and underlying mechanisms.

An ovalbumin-induced mouse AR model and immortalized human nasal epithelial cells (HNEpC) were used to assess MSC-sEV effects. Characterization and nanoparticle tracing analysis revealed that MSC-sEVs had an average diameter of 125 nm, with a size distribution between 87 and 200 nm (Fig 1A). Transmission electron microscopy confirmed their spherical morphology (Fig 1B). Western blotting showed enrichment of classical exosomal markers CD63 and CD9, and absence of the negative marker calnexin, compared to cell lysate (Fig 1C). To assess uptake by nasal epithelial cells, PKH26-labeled MSC-sEVs were cocultured with HNEpCs. Increasing PKH26 fluorescence intensity over time confirmed active uptake (Fig 1D). An experimental allergic rhinitis (AR) model was established via OVA intraperitoneal injection and nasal drops (Fig 1E). Western blot analysis showed reduced tight junction proteins (occludin, claudin-1) and mucoprotein MUC1, indicating barrier disruption, while elevated plasma IgE confirmed successful allergic sensitization (Fig 1F, G). In vivo, PKH26-labeled MSC-sEVs delivered intranasally produced positive fluorescence signals in nasal mucosa (Fig 1H). Collectively, they characterized MSC-sEVs, established an AR mouse model, and confirmed active MSC-sEV uptake by epithelial cells both in vitro and in vivo.