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Electrophoresis as a simple method to detect deleterious actions of engineered nanoparticles on living cells

Abstract : 17 The release of engineered nanoparticles (NPs) in the environment may have profound implications 18 for the health of aquatic biota. In this study, we evidence that the initial stage of NPs action on 19 bacteria can be detected upon measurement of the electrophoretic fingerprints of mixed NPs-cells 20 dispersions. Such electrokinetic signatures reflect a modification of the physicochemical cells and NPs 21 surface properties following changes in cell envelope organization, subsequent intracellular material 22 release and/or biomolecules cell excretion. The demonstration is based on a thorough investigation 23 of the electrohydrodynamic features of genetically engineered Escherichia coli bacteria exhibiting 24 distinct surface phenotypes (presence of adhesive YeeJ large proteins or F-pili proteinaceous 25 filaments) exposed to silica NPs (65 nm radius) functionalized by-NH 2 terminal groups. At pH 7, 26 electrostatics prevents interactions between bacteria and SiNH 2 NPs, regardless of the NPs 27 concentration considered (0-10-2 g L-1 range). At pH 3, electrostatically-driven interactions allow 28 intimate contacts between NPs and bacteria. In turn, significant modulation of the electrophoretic 29 determinants of cells and NPs are generated due to cells envelope alteration and bio-corona 30 acquisition by NPs. Differentiated roles of the cell surface appendages in the mediation of NPs 31 impacts are evidenced by the measured dependence of electropherograms on cell surface 32 phenotype and NPs concentration. Cell morphology and surface roughness, evaluated by Atomic 33 Force Microscopy (AFM) in liquid, confirm the pH and NPs concentration conditions where NPs-cells 34 interactions are operational. Combination of electrokinetics and AFM further pinpoints 35 heterogeneities in cells response at the single cell and population scales. Altogether, results show 36 that electrokinetics is suitable to detect the preliminary stage of events leading to NPs toxicity 37 toward microorganisms. 38 Environmental context: 39 Attractive interactions and subsequent contacts between NPs and microorganisms are the first steps 40 of a chain of events leading to adverse NPs effects toward cells. Herein we show that the 41 electrophoretic response of complex mixtures of engineered NPs (silica NPs) and bacteria (E. coli) 42 Page 2/27 reflects initial NPs-mediated cell surface damages. We thus provide a promising option for a rapid 43 electrokinetics-based detection of deleterious actions of NPs on biological cells. 44
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Elise Vouriot, Isabelle Bihannic, Audrey Beaussart, Yves Waldvogel, Angelina Razafitianamaharavo, et al.. Electrophoresis as a simple method to detect deleterious actions of engineered nanoparticles on living cells. Environmental Chemistry, CSIRO Publishing, 2019, 17, pp.39-53. ⟨10.1071/EN19190⟩. ⟨hal-02351027⟩



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