Microfluidic-assisted analysis of extracellular vesicles for advanced biomedical applications

Extracellular vesicles are lipid bilayer-enclosed nanoparticles that play a crucial role in intercellular communication by transporting bioactive molecules across cells, tissues, and even organisms. Although certain aspects of EV biogenesis, function, and metabolic pathways are yet to be fully elucidated, their distinctive features such as cargo protection, structural stability, and the capacity to cross biological barriers render them highly promising for medical applications, especially as novel biomarkers for disease diagnosis. However, the ultrasmall size and complex surface properties pose substantial challenges to their analysis. Microfluidic technology has emerged as a powerful tool for EV isolation and analysis. It offers remarkable advantages, including high throughput, efficiency, and minimal sample consumption, providing innovative solutions for isolating and analyzing EVs with unparalleled precision. Moreover, the integration of artificial intelligence and advanced digital analytics with microfluidic platforms has opened new avenues for enhanced data processing and diagnostic accuracy. In this review, we comprehensively summarize recent advances in EV research, beginning with an in-depth analysis of EVs' biogenesis mechanisms and the formation of disease-specific biomarkers. Then EV-mediated pathological processes with diagnostic significance are discussed, followed by a comprehensive evaluation of microfluidic-based approaches for isolation, purification, and single-vesicle analysis of EVs. The clinical diagnostic potential of EVs is assessed across major disease categories including cancers, cardiovascular disorders, neurological conditions, infectious diseases, and autoimmune pathologies, highlighting their emerging role as next-generation biomarkers. Finally, the current advances in EV-based diagnostics are discussed, along with the future perspectives on microfluidic-based EV analysis towards various biomedical applications and clinical translation.