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New insight into artifactual phenomena during in vitro toxicity assessment of engineered nanoparticles: Study of TNF-α adsorption on alumina oxide nanoparticle

Abstract : Biomolecules can be adsorbed on nanoparticles (NPs) and degraded during in vitro toxicity assays. These artifactual phenomena could lead to misinterpretation of biological activity, such as false-negative results. To avoid possible underestimation of cytokine release after contact between NP and cells, we propose a methodology to account for these artifactual phenomena and lead to accurate measurements. We focused on the pro-inflammatory cytokine tumor necrosis factor TNF-α. We studied well-characterized boehmite engineered NP [aluminum oxide hydroxide, AlO(OH)]. The rate of TNF-α degradation and its adsorption (on boehmite and on the walls of wells) were determined in cell-free conditions by adding a known TNF-α concentration (1500 pg/ml) under various experimental conditions. After a 24-h incubation, we quantified that 7 wt.% of the initial TNF-α was degraded over time, 6 wt.% adsorbed on the walls of 96-well plates, and 13 wt.% adsorbed on the boehmite surface. Finally, boehmite NP were incubated with murine macrophages (RAW 264.7 cell line). The release of TNF-α was assessed for boehmite NP and the experimental data were corrected considering the artifactual phenomena, which accounted for about 20-30% of the total
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Submitted on : Monday, March 11, 2013 - 4:31:26 PM
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Mélanie Pailleux, Delphine Boudard, Jérémie Pourchez, Valérie Forest, Philippe Grosseau, et al.. New insight into artifactual phenomena during in vitro toxicity assessment of engineered nanoparticles: Study of TNF-α adsorption on alumina oxide nanoparticle. Toxicology in Vitro, Elsevier, 2013, 27, pp.1049-1056. ⟨10.1016/j.tiv.2013.01.022⟩. ⟨hal-00799129⟩

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