Internalization, cytotoxicity, oxidative stress and inflammation of multi-walled carbon nanotubes in human endothelial cells: influence of pre-incubation with bovine serum albumin

Abstract

When entering circulation, multi-walled carbon nanotubes (MWCNTs) will inevitably adsorb proteins, which can consequently influence their toxicity to cells lining human blood vessels. In this study, we investigated the influence of pre-incubation with bovine serum albumin (BSA) on internalization, cytotoxicity, oxidative stress and inflammation induced by pristine/carboxylated MWCNTs to human umbilical vein endothelial cells (HUVECs). Atomic force microscopy (AFM) indicated the adsorption of proteins onto the surface of MWCNTs, which consequently increased the diameter. Pre-incubation with BSA did not obviously influence the hydrodynamic sizes, but decreased the zeta potential of MWCNTs. Transmission electron microscopy (TEM) indicated the internalization of both types of MWCNTs into HUVECs, whereas pre-incubation with BSA appeared to enhance the internalization. MWCNT exposure induced cytotoxicity and oxidative stress, as well as a modest inflammatory response shown as an increased THP-1 adhesion to HUVECs, but not release of interleukin 6 (IL-6) or tumor necrosis factor (TNFα). Exposure to MWCNTs pre-incubated with BSA induced less cytotoxicity to HUVECs, associated with increased intracellular glutathione (GSH). However, MWCNTs induced IL-6 and TNFα release, as well as THP-1 adhesion to HUVECs, were enhanced after pre-incubation with BSA. In summary, these data indicated that pre-incubation with BSA could enhance the internalization of MWCNTs to HUVECs, which consequently reduces the cytotoxicity and oxidative stress, but enhances the inflammatory response of MWCNTs. The reduced cytotoxicity and oxidative stress, and enhanced inflammatory responses are likely due to a combined effect of BSA and MWCNTs, which suggests that when assessing the biological effects of MWCNTs in circulation, it is necessary to consider the interactions between MWCNTs and serum proteins.