Protein Corona Formation on Soft Nanocarriers Under Biomimetic Circulatory Flow

ABSTRACT Upon exposure to biological fluids, nanoparticles are rapidly coated by a layer of adsorbed proteins known as the protein corona (PC). PC composition strongly influences the nano‐bio interactions, yet its formation under physiologically relevant flow remains insufficiently explored. Here, a venous circulation‐mimicking system is designed to replicate physiological shear forces and dynamic protein exchange encountered during intravenous administration. PC formation on commonly utilized soft nanocarriers, including liposomes, poly(lactic‐ co ‐glycolic acid) nanoparticles (PLGA NPs), and lipid‐polymer hybrid nanoparticles (LPNPs) is investigated under biomimetic flow. The results demonstrate that flow‐induced PC formation alters the physicochemical properties of nanocarriers in a nanocarrier type‐dependent manner. Proteomic PC profiling reveals a largely shared protein core among all nanocarriers, alongside distinct material‐specific protein signatures. Uptake studies in A549 cells show that PC formation markedly reduces nanocarrier uptake for all formulations, indicating that pre‐formed coronas persistently modulate nanoparticle‐cell interactions. These findings highlight the central role of PC formation in defining nanocarrier biological identity and demonstrate that biomimetic circulation modulates corona organization without overriding serum‐driven effects. This study highlights the importance of incorporating dynamic and physiologically relevant conditions into in vitro models to better understand and evaluate nanocarrier behavior, and guide the rational design of nanomedicine platforms.