title: 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.
authors: LINA, EA 4624, F-42023 Saint-Etienne, France. pailleux@emse.fr
authors: Mélanie
authors: M
authors: Pailleux
authors: Delphine
authors: D
authors: Boudard
authors: Jérémie
authors: J
authors: Pourchez
authors: Valérie
authors: V
authors: Forest
authors: Philippe
authors: P
authors: Grosseau
authors: Michèle
authors: M
authors: Cottier
mesh_terms: D000327:Adsorption; D000536:Aluminum Hydroxide / Q000737:chemistry* / Q000633:toxicity; D000537:Aluminum Oxide / Q000737:chemistry* / Q000633:toxicity; D000818:Animals; D016477:Artifacts; D002460:Cell Line; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D051379:Mice; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles / Q000737:chemistry* / Q000633:toxicity / Q000648:ultrastructure; D018675:Toxicity Tests; D014409:Tumor Necrosis Factor-alpha / Q000737:chemistry* / Q000378:metabolism
chemical_list: D014409:Tumor Necrosis Factor-alpha; D000536:Aluminum Hydroxide; C069471:aluminum oxide hydroxide; D007770:L-Lactate Dehydrogenase; D000537:Aluminum Oxide