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Etude de l'intéraction nanoparticules-bactéries : application à l'élaboration d'un biocapteur

Abstract : Despite the growing enthusiasm for nanotechnologies, nanoparticles (NPs) might put environmental safety and human health at risk, as they can interact with biological systems and affect their behavior. It is therefore essential to know their mechanisms of interactions in order not only to prevent their potential risks but also to benefit from their unique properties, such as in biosensors design. In this context, we study the cytotoxicity of silica NPs, with diverse sizes and charges, on the properties of Escherichia coli and Bacillus subtilis bacteria, by means of atomic force microscopy and viability tests. Negatively charged NPs (NPs-) with a diameter φ lower than a critical diameter φc, 50 - 80 nm, (i) lead to the isolation of E. coli bacteria, (ii) induce a "spherification" of the cell initially rod shaped, and (iii) cause the formation of pore-like lesions in the outer membrane and a reorganization of its structure. For B. subtilis bacteria, only the degradation of the peptidoglycane’s structure was observed. Though, for both strains, an antibacterial activity was shown for NPs- below φc, which potentially lead to the cell lysis whereas, above φc, NPs- have no effect on population, morphology or bacterial structure. As positively charged NPs are concerned, whatever their diameter, they lead to a strong aggregation of the cells, due to electrostatic interactions, and tend to favor the formation of membrane invaginations, not necessarily involving cell lysis. This fundamental study has been used to develop an electrochemical biosensor for bacteria, which are of great importance for biomedical, environmental and defense issues. NPs involved in such tools offer a fast, high-sensitive and low-cost way of detection. A polyelectrolyte multilayer was used to immobilize harmless NPs (φ = 100 nm), which are, then, functionalized with specific antibodies, in order to enhance the final detection of E. coli bacteria. All steps were optimized by a spin coating process and studied through quartz microbalance and cyclic voltametry measurements. Integrating NPs in this biosensor resulted in a linear and unsaturated detection of E. coli bacteria in a wide range of concentration (until 10^9 CFU/mL) and a limit of detection of 10^6 CFU/mL.
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Submitted on : Tuesday, December 12, 2017 - 12:35:14 AM
Last modification on : Friday, October 19, 2018 - 1:36:05 PM

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Marion Mathelié-Guinlet. Etude de l'intéraction nanoparticules-bactéries : application à l'élaboration d'un biocapteur. Chimie théorique et/ou physique. Université de Bordeaux, 2017. Français. ⟨NNT : 2017BORD0697⟩. ⟨tel-01661488⟩

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