Quantification of C60-induced membrane disruption using a quartz crystal microbalance

Abstract

Direct contact between fullerene C₆₀ nanoparticles (NPs) and cell membranes is one of mechanisms for its cytotoxicity. In this study, the influence of C₆₀ NPs on lipid membranes was investigated. Giant unilamellar vesicles (GUVs) were used as model cell membranes to observe the membrane disruption after C₆₀ exposure. C₆₀ NPs disrupted the positively charged GUVs but not the negatively charged vesicles, confirming the role of electrostatic forces. To quantify the C₆₀ adhesion on membrane and the induced membrane disruption, a supported lipid bilayer (SLB) and a layer of small unilamellar vesicles (SUVs) were used to cover the sensor of a quartz crystal microbalance (QCM). The mass change on the SLB (Δm_SLB) was caused by the C₆₀ adhesion on the membrane, while the mass change on the SUV layer (Δm_SUV) was the combined result of C₆₀ adhesion (mass increase) and SUV disruption (mass loss). The surface area of SLB (A_SLB) was much smaller than the surface area of SUV (A_SUV), but Δm_SLB was larger than Δm_SUV after C₆₀ deposition, indicating that C₆₀ NPs caused remarkable membrane disruption. Therefore a new method was built to quantify the degree of NP-induced membrane disruption using the values of Δm_SUV/Δm_SLB and A_SUV/A_SLB. In this way, C₆₀ can be compared with other types of NPs to know which one causes more serious membrane disruption. In addition, C₆₀ NPs caused negligible change in the membrane phase, indicating that membrane gelation was not the mechanism of cytotoxicity for C₆₀ NPs. This study provides important information to predict the environmental hazard presented by fullerene NPs and to evaluate the degree of membrane damage caused by different NPs.