Engineering Biomimetic 3D Microenvironments for Extracellular Vesicle Programming Toward Clinical Translation

The cell secretome includes extracellular vesicles (EVs), nanoscale lipid bilayer-enclosed particles that carry diverse bioactive cargos, including proteins, lipids, and nucleic acids. As key mediators of paracrine signaling, EVs reflect the molecular and functional characteristics of their parent cells and play critical roles in regulating tissue homeostasis and regeneration. Growing evidence supports their therapeutic potential across a wide range of diseases. However, the clinical translation of EV-based therapies remains limited by challenges related to yield, purity, targeting specificity, and functional consistency. Recent advances in biomimetic culture systems—particularly three-dimensional (3D) platforms that recapitulate features of the native extracellular matrix microenvironment—have demonstrated a strong influence on cell phenotype, secretory activity, and EV composition. This review highlights how biochemical and mechanical cues within 3D culture systems regulate EV biogenesis, cargo loading, and functional outcomes and discusses their implications for improving the scalability, efficacy, and clinical translation of EV-based therapeutics.