Myeloperoxidase (MPO) from activated neutrophils plays important roles in multiple human inflammatory diseases by catalyzing the formation of powerful oxidant hypochlorous acid (HOCl). As a major flavonoid in the human diet, quercetin has been suggested to act as antioxidant and anti-inflammatory agent in vitro and in vivo. In this study, we showed that quercetin inhibited MPO-mediated HOCl formation (75.0 ± 6.2% for 10 μM quercetin versus 100 ± 5.2% for control group, P < 0.01) and cytotoxicity to endothelial cells in vitro, while this flavonoid was nontoxic to endothelial cell cultures ( P > 0.05, all cases). Moreover, quercetin inhibited HOCl generation by stimulated neutrophils (a rich source of MPO) and protected endothelial cells from neutrophils-induced injury. Furthermore, quercetin could inhibit HOCl-induced endothelial dysfunction such as loss of cell viability, and decrease of nitric oxide formation in endothelial cells ( P < 0.05, all cases). Consistent with these in vitro data, quercetin attenuated lipopolysaccharide-induced endothelial dysfunction and increase of MPO activity in mouse aortas, while this flavonoid could protect against HOCl-mediated endothelial dysfunction in isolated aortas ( P < 0.05). Therefore, it was proposed that quercetin attenuated endothelial injury in inflammatory vasculature via inhibition of vascular-bound MPO-mediated HOCl formation or scavenging of HOCl. These data indicate that quercetin is a nontoxic inhibitor of MPO activity and MPO/neutrophils-induced cytotoxicity in endothelial cells and may be useful for targeting MPO-dependent vascular disease and inflammation.

The interactions between proteins and bioactive ligands (such as flavonoids and nanomaterials) are vital to the design of effective protein carriers for the protection of bioactive molecules and reduction of the cytotoxicity of nanotubes. Bovine serum albumin (BSA) can bind various bioactive components and subsequently form protein-ligand complexes. Herein, the binding of BSA to quercetin and single-walled carbon nanotubes (SWCNTs) was investigated by using experimental and molecular-docking methods. The fluorescence intensity of BSA was decreased by both quercetin and SWCNTs in static quenching mode (i.e., compound formation), which was authenticated by Stern-Volmer calculations. Although quercetin showed a higher affinity for BSA than SWCNTs, the binding of both components to BSA was located in site I (subdomain IIA). BSA-diligand complexes were successfully generated when SWCNTs and quercetin, in that sequence, were added. The cytotoxicity of SWCNTs and the formation of reactive oxygen species in endothelial cells were decreased with the BSA-diligand complexes relative to those of SWCNTs or BSA-SWCNT corona, whereas the stability problems of quercetin were ameliorated in the BSA-diligand complex relative to in the free flavonoid. The BSA-diligand complex showed a better inhibitive effect on the cytotoxicity of SWCNTs than the BSA-SWCNT complex, and thus the coexistence of quercetin played a crucial role. These data demonstrate the advantages and possibility of designing BSA carriers for the protection of bioactive ligands and reduction of the cytotoxicity of nanotubes in functional-food and biomedical applications.

Proteins in the bloodstream bind to carbon nanotubes (CNTs) through noncovalent interactions to form a protein corona, thereby effectively influencing the biological properties and blood biocompatibility of the CNTs. Here, we investigated the binding of common plasma proteins (i.e., human immunoglobulin G (IgG), human serum albumin (HSA), and fibrinogen (FG)) to carboxylated single-walled CNTs (SWCNTs), and evaluated the effects of these different protein coronas on cytotoxicity to endothelial cells and immune response to neutrophils in the bloodstream. Measurements of adsorption parameters revealed tight binding of proteins to SWCNTs, and the SWCNTs adsorption capacities followed the order FG > HSA > IgG. In addition, the basic residues (Arg, Lys, His) were found to play an important role in the formation of protein-SWCNTs corona complexes and determine their adsorption capacity. Consistent with the higher protein adsorption capacity, FG more significantly reduced the cytotoxicity of CNTs to human umbilical vein endothelial cells than the other two proteins. However, only treatment of SWCNTs with IgG resulted in the enhancement of CNT-induced myeloperoxidase (MPO) release (i.e., neutrophil activation) in neutrophils, while MPO-dependent degradation of CNTs induced less cytotoxicity than initial nanomaterials. Consistent with these effects of protein coronas, the presence of serum attenuated the cytotoxicity of CNTs and CNTs could induce neutrophil activation in human blood plasma. Our study demonstrates the ability of adsorbed plasma proteins to influence cytotoxicity and neutrophil response caused by CNTs in the bloodstream.

Carbon nanotubes are widely used in the area of biomedicine, and the binding of protein to carbon nanotubes are believed to play an important role in the potential cytotoxicity of nanomaterials. In this work, we investigated the effects of human fibrinogen-surface coatings on the biodegradation and cytotoxicity of carboxylated single-walled carbon nanotubes (SWCNTs). It was found that the electrostatic and π-π stacking interactions might be the crucial factors in stabilizing the binding of fibrinogen with SWCNTs by both theoretical and experimental approaches. Although naked SWCNTs could induce significant toxicity to macrophages, coating these nanomaterials with fibrinogen could greatly attenuate their toxicity. On the other hand, although SWCNTs and fibrinogen-preincubated SWCNTs were resistant to biodegradation in resting macrophages, both naked and fibrinogen-coated SWCNTs could be effectively and similarly degraded through myeloperoxidase (MPO) and peroxynitrite (ONOO)-dependent pathways in activated macrophages, where NADPH oxidase played a determinant role in the biodegradation process. Importantly, degraded SWCNTs by ONOO pathway in vitro induced less cytotoxicity than non-degraded nanotubes. These findings demonstrated that the binding of fibrinogen to SWCNTs could reduce cytotoxicity without affecting the biodegradation of nanotubes in activated inflammatory cells, providing a new route to design the safer nanotubes for future biomedical applications.