Supported Erythrocyte Membranes on Piezoelectric Sensors for Studying the Interactions with Nanoparticles

Applications of nanoparticles (NPs) in nanodrugs, food additives, and cosmetics can result in the presence of nanomaterials in the human circulatory system and their attachment to red blood cells (RBCs), which may lead to cytotoxic effects. To investigate the interactions of NPs with RBC membranes (RBCm), supported erythrocyte membranes (SRBCm) were developed on piezoelectric sensors in a quartz crystal microbalance with dissipation (QCM-D) at 25 °C. A well-dispersed RBCm suspension at 1 mM NaCl and 0.2 mM NaHCO₃ was obtained from whole blood and comprised colloidal membrane fragments with the average hydrodynamic diameter and zeta potential of 390 nm and -0.53 mV, respectively, at pH 7.0. The thin and rigid SRBCm was formed mainly through the deposition of RBCm fragments on the poly-l-lysine-modified crystal sensor, leading to the average frequency shift of -26.2 Hz and the low ratio of the dissipation to frequency shift (7.2 × 10^-8 Hz^-1). The complete coverage of SRBCm was indicated by the plateau of the frequency shift in the stage of SRBCm formation and no deposition of negatively charged 106 nm polystyrene nanoparticles (PSNPs) on the SRBCm. Atomic force microscopy and immunofluorescence microscopy images showed that RBCm aggregates with the average size of 420 nm and erythrocyte membrane proteins existed on SRBCm, respectively. The methods of determining attachment efficiencies of model positively charged NPs (i.e., hematite NPs or HemNPs) and model negatively charged NPs (i.e., PSNPs) on SRBCm were demonstrated in 1 mM NaCl solution at pH 5.1 and pH 7.0, respectively. HemNPs exhibited a favorable deposition with an attachment efficiency of 0.99 while PSNPs did not show any attachment propensity toward SRBCm.