Characterization of Exosome-like Nanoparticles from Saffron Tepals and Their Immunostimulatory Activity

Plant exosomes exhibit high stability and easy absorption, and have emerged as promising bioactive tools due to their potential health benefits and biomedical applications. Saffron tepals contain abundant metabolites with potential therapeutic properties and were used for exosome extraction by ultracentrifugation and gradient purification. The exosomes showed an average particle size of 151.5 ± 79.6 nm and exhibited a spherical morphology. Five well-conserved miRNAs-miR157, miR166, miR168, miR396, and miR398-were identified in the exosomes, which are involved in the coordination of growth and physiological plant responses with endogenous and environmental abiotic and biotic signals, and their potential targets in mammals are upregulated in specific cancer types and associated with inflammation. Proteome analysis revealed an enrichment of proteasome proteins, ribosomal proteins, and proteins involved in the cytoskeleton, transport across the membrane (ABC transporters), and vesicle trafficking (RAB GTPases, TM9SF and Coatomer subunits). Metabolite analyses showed mainly anthocyanins. The exosomes have selective stimulatory activity on macrophages, increasing the expression of surface molecules (<i>CD80</i> and <i>CD86</i>), and cytokines (<i>IL-1β</i>, <i>IL-6</i>, and <i>TNF-α</i>), but not the levels of <i>IL-10</i>. Overall, these results indicated that saffron flowers are an effective and abundant source of exosomes as new nanomedicines for human health.