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The molecular responses of macrophages to copper-based nanoparticles havebeen investigated via a combination of proteomic and biochemical approaches,using the RAW264.7 cell line as a model. Both metallic copper and copper oxidenanoparticles have been tested, with copper ion and zirconium oxidenanoparticles used as controls. Proteomic analysis highlighted changes inproteins implicated in oxidative stress responses (superoxide dismutases andperoxiredoxins), glutathione biosynthesis, the actomyosin cytoskeleton, andmitochondrial proteins (especially oxidative phosphorylation complex subunits).Validation studies employing functional analyses showed that the increases inglutathione biosynthesis and in mitochondrial complexes observed in theproteomic screen were critical to cell survival upon stress with copper-basednanoparticles; pharmacological inhibition of these two pathways enhanced cellvulnerability to copper-based nanoparticles, but not to copper ions.Furthermore, functional analyses using primary macrophages derived from bonemarrow showed a decrease in reduced glutathione levels, a decrease in themitochondrial transmembrane potential, and inhibition of phagocytosis and oflipopolysaccharide-induced nitric oxide production. However, only a fraction ofthese effects could be obtained with copper ions. In conclusion, this studyshowed that macrophage functions are significantly altered by copper-basednanoparticles. Also highlighted are the cellular pathways modulated by cellsfor survival and the exemplified cross-toxicities that can occur betweencopper-based nanoparticles and pharmacological agents.

Molecular responses of mouse macrophages to copper and copper oxide nanoparticles inferred from proteomic analyses