To maintain osmotic balance, cells usually produce neutral solutes (i.e., osmolytes), together with charged species to cope with salinity stress. Osmolytes are known to be important in stabilizing/destabilizing macromolecules (e.g., proteins) via depletion/accumulation around their surfaces. To better understand the physiological fate of nanoparticles (NPs), we investigated the effect of osmolytes [(urea and trimethylamine N-oxide (TMAO)] and specific anions (NO3- and F-) on the interactions between NPs and supported lipid bilayers (SLBs). Carboxylated polystyrene NPs (60 nm) and 1,2-dioleoyl-sn-glycero3-phosphocholine (DOPC) were chosen as model NPs and lipid. Quartz crystal microbalance with dissipation monitoring (QCM-D) was used to quantify NP deposition dynamics. Microscale thermophoresis (MST) was used to characterize the affinity between DOPC vesicles (or NPs) and osmolytes. Our results show that osmolytes are capable of protecting SLBs from NP-induced disruption. Upon NP deposition onto supported vesicle layers (SVLs), the leakage of encapsulated dyes decreased with the addition of osmolytes. The combination of kosmotropes (TMAO and F-) are more efficient than that of chaotropes (urea and NO3-) in weakening the hydrophobic interaction between NPs and SLBs by preferential binding to NPs and/or SLBs. (C) 2020 Elsevier Inc. All rights reserved.