S. Ghosal, Rita Leporati, Árpád Varga, Petra Susánszki, Nóra Fekete, Tamás László, Tünde Bárkai, Fülöp Károly Grébecz, Delaram Khamari, Tárek Zoltán Magyar, Marcus Höring, Krisztina V Vukman, Bernadett R. Bodnár, Brachyahu Meir Kestecher, Mohamed Abdel Fattah, Csaba Bödör, József Maléth, Gerhard Liebisch, Evelyn Orsó, Edit I. Buzás, Xabier Osteikoetxea
Conclusion These findings provide a robust compositional framework to distinguish non-vesicular extracellular particles from canonical EVs and underscore their emerging significance as mediators of intercellular communication and potential vehicles for targeted delivery.
Abstract Background Mechanisms governing non-vesicular extracellular particle (NVEP) biogenesis and compositional diversity remain incompletely characterized. Method This study integrates multiparametric profiling to define NVEPs. NVEPs were isolated by differential ultracentrifugation and characterized using complementary biochemical, lipidomic, and imaging approaches, including protein-to-lipid ratio analysis, lipid profiling, and advanced microscopy. Result Comprehensive characterization demonstrated that NVEPs exhibit significantly elevated protein-to-lipid ratios compared to extracellular vesicles (EVs) subtypes. Lipidomic profiling revealed pronounced enrichment in cholesteryl esters and triacylglycerols, with distinct carbon chain length signatures differentiating NVEPs from small EVs. Super-resolution microscopy confirmed marker heterogeneity across populations, with NVEPs showing specific enrichment of Arf6 and CD63. Conclusion These findings provide a robust compositional framework to distinguish non-vesicular extracellular particles from canonical EVs and underscore their emerging significance as mediators of intercellular communication and potential vehicles for targeted delivery.