Ibtissem Gammoudi

Love-wave bacteria-based sensor for the detection of heavy metal toxicity in liquid medium

The present work deals with the development of a Love-wave bacteria-based sensor platform for the detection of heavy metals in liquid medium. The acoustic delay-line is inserted in an oscillation loop in order to record the resonance frequency in real-time. A Polydimethylsiloxane (PDMS) chip with a liquid chamber is maintained by pressure above the acoustic wave propagation path. Bacteria (Escherichia coli) were fixed as bioreceptors onto the sensitive surface of the sensor coated with a polyelectrolyte (PE) multilayer using a simple and efficient layer-by-layer (LbL) electrostatic self-assembly procedure. Poly(allylamine hydrochloride) (PAH cation) and poly(styrene sulfonate) (PSS anion) were alternatively deposited so that the strong attraction between oppositely charged polyelectrolytes resulted in the formation of a (PAH-PSS)(n)-PAH molecular multilayer. The real-time characterization of PE multilayer and bacteria deposition is based on the measurement of the resonance frequency perturbation due to mass loading during material deposition. Real-time response to various concentrations of cadmium (Cd(2+)) and mercury (Hg(2+)) has been investigated. A detection limit as low as 10(-12) mol/l has been achieved, above which the frequency increases gradually up to 10(-3) mol/l, after a delay of 60 s subsequent to their introduction onto bacterial cell-based biosensors. Beyond a 10(-3) mol/l a steep drop in frequency was observed. This response has been attributed to changes in viscoelastic properties, related to modifications in bacteria metabolism.

High sensitive mesoporous TiO2-coated Love wave device for heavy metal detection

This work deals with the design of a highly sensitive whole cell-based biosensor for heavy metal detection in liquid medium. The biosensor is constituted of a Love wave sensor coated with a polyelectrolyte multilayer (PEM). Escherichia coli bacteria are used as bioreceptors as their viscoelastic properties are influenced by toxic heavy metals. The acoustic sensor is constituted of a quartz substrate with interdigitated transducers and a SiO2 guiding layer. However, SiO2 shows some degradation when used in a saline medium. Mesoporous TiO2 presents good mechanical and chemical stability and offers a high active surface area. Then, the addition of a thin titania layer dip-coated onto the acoustic path of the sensor is proposed to overcome the silica degradation and to improve the mass effect sensitivity of the acoustic device. PEM and bacteria deposition, and heavy metal influence, are real time monitored through the resonance frequency variations of the acoustic device. The first polyelectrolyte layer is inserted through the titania mesoporosity, favouring rigid link of the PEM on the sensor and improving the device sensitivity. Also, the mesoporosity of surface increases the specific surface area which can be occupied and favors the formation of homogeneous PEM. It was found a frequency shift near -20±1 kHz for bacteria immobilization with titania film instead of -7±3 kHz with bare silica surface. The sensitivity is highlighted towards cadmium detection. Moreover, in this paper, particular attention is given to the immobilization of bacteria and to biosensor lifetime. Atomic Force Microscopy characterizations of the biosurface have been done for several weeks. They showed significant morphological differences depending on the bacterial life time. We noticed that the lifetime of the biosensor is longer in the case of using a mesoporous TiO2 layer.

Probing the threshold of membrane damage and cytotoxicity effects induced by silica nanoparticles in Escherichia coli bacteria

The engineering of nanomaterials, because of their specific properties, is increasingly being developed for commercial purposes over the past decades, to enhance diagnosis, cosmetics properties as well as sensing efficiency. However, the understanding of their fate and thus their interactions at the cellular level with bio-organisms remains elusive. Here, we investigate the size- and charge-dependence of the damages induced by silica nanoparticles (SiO2-NPs) on Gram-negative Escherichia coli bacteria. We show and quantify the existence of a NPs size threshold discriminating toxic and inert SiO2-NPs with a critical particle diameter (Φc) in the range 50nm-80nm. This particular threshold is identified at both the micrometer scale via viability tests through Colony Forming Units (CFU) counting, and the nanometer scale via atomic force microscopy (AFM). At this nanometer scale, AFM emphasizes the interaction between the cell membrane and SiO2-NPs from both topographic and mechanical points of view. For SiO2-NPs with Φ>Φc no change in E. coli morphology nor its outer membrane (OM) organization is observed unless the NPs are positively charged in which case reorganization and disruption of the OM are detected. Conversely, when Φ

Influence of oxidized lipids on palmitoyl-oleoyl-phosphatidylcholine organization, contribution of Langmuir monolayers and Langmuir–Blodgett films

In this work, we studied the interaction of two oxidized lipids, PoxnoPC and PazePC, with POPC phospholipid. Mean molecular areas obtained from (π-A) isotherms of mixed PoxnoPC-POPC and PazePC-POPC monolayers revealed different behaviors of these two oxidized lipids: the presence of PoxnoPC in the monolayers induces their expansion, mean molecular areas being higher than those expected in the case of ideal mixtures. PazePC-POPC behave on the whole ideally. This difference can be explained by a different conformation of oxidized lipids. Moreover the carboxylic function of PazePC is protonated under our experimental conditions, as shown by (π-A) isotherms of PazePC at different pH values. Both oxidized lipids induce also an increase of the monolayer elasticity, PoxnoPC being slightly more efficient than PazePC. These monolayers were transferred from the air-water interface onto mica supports for a study by AFM. AFM images are on the whole homogenous, suggesting the presence of only one lipid phase in both cases. However, in the case of PazePC-POPC monolayers, AFM images show also the presence of areas thicker of 7nm to 10nm than the surrounding lipid phase, probably due to the local formation of multilayer systems induced by compression.