Sandra Casale

Sensing the Charge State of Single Gold Nanoparticles via Work Function Measurements

Electrostatic interactions at the nanoscale can lead to novel properties and functionalities that bulk materials and devices do not have. Here we used Kelvin probe force microscopy (KPFM) to study the work function (WF) of gold nanoparticles (NPs) deposited on a Si wafer covered by a monolayer of alkyl chains, which provide a tunnel junction. We find that the WF of Au NPs is size-dependent and deviates strongly from that of the bulk Au. We attribute the WF change to the charging of the NPs, which is a consequence of the difference in WF between Au and the substrate. For an NP with 10 nm diameter charged with ∼ 5 electrons, the WF is found to be only ∼ 3.6 eV. A classical electrostatic model is derived that explains the observations in a quantitative way. We also demonstrate that the WF and charge state of Au NPs are influenced by chemical changes of the underlying substrate. Therefore, Au NPs could be used for chemical and biological sensing, whose environmentally sensitive charge state can be read out by work function measurements.

Optimizing the immobilization of gold nanoparticles on functionalized silicon surfaces: amine- vs thiol-terminated silane

Immobilization of gold nanoparticles on planar surfaces is of great interest to many scientific communities; chemists, physicists, biologists, and the various communities working at the interfaces between these disciplines. Controlling the immobilization step, especially nanoparticles dispersion and coverage, is an important issue for all of these communities. We studied the parameters that can influence this interaction, starting with the nature of the terminal chemical function. Thus, we have carefully grafted silanes terminated by either amine or thiol groups starting from aminopropyltriethoxysilane (APTES) or mercaptopropyltriethoxysilane. We also changed the chain length for thiol-terminated layers through covalent grafting of mercaptoundecanoic acid (MUA) on APTES-modified layers, and the protocol of nanoparticles deposition to evaluate whether other factors must be taken into consideration to rationalize this interaction. The formed layers were characterized by X-ray photoelectron spectroscopy and gold nanoparticles deposition was monitored by scanning electron microscopy and surface-enhanced Raman scattering. We observed significant differences in terms of nanoparticles dispersion and density depending on the nature of the chemical layer on silicon. The use of ultrasounds during the deposition process was very efficient to limit aggregates formation. The optimal deposition procedures were obtained through the use of APTES and APTES/MUA functionalization. They were compared in terms of coverage, dispersion, and densities of isolated nanoparticles. The APTES/MUA surfaces clearly showed better results that may arise from both the longer chain and the dilution of thiol end groups.

The amphiphilic hydrophobin Vmh2 plays a key role in one step synthesis of hybrid protein-gold nanoparticles.

We report a simple and original method to synthesize gold nanoparticles in which a fungal protein, the hydrophobin Vmh2 from Pleurotus ostreatus and dicarboxylic acid-terminated polyethylene-glycol (PEG) has been used as additional components in a one step process, leading to hybrid protein-metal nanoparticles (NPs). The nanoparticles have been characterized by ultra-violet/visible, infrared and X-ray photoelectron spectroscopies, dynamic light scattering and also by electron microscopy imaging. The results of these analytical techniques highlight nanometric sized, stable, hybrid complexes of about 12 nm, with outer surface rich in functional chemical groups. Interaction with protein and antibodies has also been exploited.

Bioconjugated gold nanorods to enhance the sensitivity of FT-SPR-based biosensors.

In this paper, we report a feasible solvent-free approach for the synthesis and self-assembling of gold nanorods after bioconjugation to antibodies; scanning electron microscopy (SEM) and transmission electron microscopy (TEM) images revealed a remarkable shape and size narrow distribution, as well as a tendency to form linear assemblies on a clean gold surface, upon interaction with antibodies. These bioconjugated gold nanorods were in turns tested in a model Fourier Transform Surface Plasmon Resonance (FT-SPR)-based immunosensor, leading to an improvement of the detection sensitivity by a factor of 8. The results highlight the simplicity of the synthesis protocol of gold nanorods and the interest of using them as labels to enhance the sensitivity of SPR-based sensors.

Selective amino acid substitution reduces cytotoxicity of the antimicrobial peptide mastoparan

The emergence of antibiotic-resistant clinical isolates and the decreased rate of development of new antibiotics are a constant threat to human health. In this context, the therapeutic value of mastoparan (MP), a toxin from wasp venom, has been extensively studied. However, since MP shows significant cytotoxic activities, further optimization is needed. Here we evaluated the antimicrobial and cytolytic activities of an MP analog created by Ala-substitution in positions 5 and 8, named [I5, R8] mastoparan ([I5, R8] MP). We found that [I5, R8] MP displayed a broad-spectrum antimicrobial activity against bacteria and fungi (MIC in the range 3-25μM), without being hemolytic or cytotoxic toward HEK-293 cells. In addition, [I5, R8] MP-amide was highly potent (MIC=3μM) against antibiotic-resistant bacteria. The interaction with microbial membranes was investigated revealing that [I5, R8] MP is able to form an active amphipathic α-helix conformation and to disturb membranes causing lysis and cell death. Based on our findings, we hypothesize that [I5, R8] MP follows a mechanism of action similar to that proposed for MP, where the pore-forming activity leads to cell death. Our results indicate that hydrophobic moment modified by amino acid substitution may enhance MP selectivity.

End-Cretaceous akaganéite as a mineral marker of Deccan volcanism in the sedimentary record

An enigmatic chloride-rich iron (oxyhydr)oxide has been recently identified together with mercury anomalies in End-Cretaceous marine sediments coeval with the Deccan Traps eruptions. The mineral was observed in Bidart (France) and Gubbio (Italy), suggesting a widespread phenomenon. However, the exact nature and origin of this Cl-bearing mineral remained speculative. Here, we characterized the accurate composition and nanostructure of this chloride-rich phase by using micro-Raman spectroscopy, Transmission (TEM) and Scanning (SEM) Electron Microscopy on Focused Ion Beam foils. We also provide new evidence of its occurrence in Zumaia, a reference KPg section from Spain. Results confirm akaganéite (β-FeOOH) as the main phase, with chloride content of 3–5 atomic weight %. Akaganéite particles are constituted by the aggregation of nanorods of akaganéite. Internal structures contain empty spaces, suggesting formation in a low-density (atmospheric) environment. This new mineralogical evidence supports the hypothesis that the observed akaganéite was formed in the Deccan volcanic plume and was transported to the Atlantic and Tethysian realms through the stratosphere. Therefore, akaganéite provides a potential new sedimentary marker to identify the imprint of the Deccan eruptions in the stratigraphic record and is evidence of volcanic halogen degassing and its potential role for the Cretaceous-Tertiary mass extinction.

Brake wear (nano)particle characterization and toxicity on airway epithelial cells in vitro

Particulate air pollution results from different sources, among which those related to road traffic have a significant impact on human health. Combustion-derived particles emitted by thermal engines have been incriminated and are now better controlled. In contrast, non-exhaust emission sources related to car wear and degradation processes are not yet regulated. Here we report on brake wear particles (BWP) harvested in two test facilities operating in France, providing samples from different braking systems and driving/testing conditions. Using a combination of light scattering, X-ray fluorescence, optical and electron microscopy, the particle size and elemental composition are revealed. The BWP are shown to be in the nano- to micrometer range and to have a low carbonaceous content (6%), iron and copper being the main components (> 40%). To evaluate the toxicity potential of its nano-sized fraction, brake wear nanoparticles are isolated by sonication, filtration and ultra-centrifugation techniques, leading to stable colloidal dispersions. A significant outcome of this study is that the nano-sized fraction represents 26% by mass of the initial BWP. Human bronchial epithelial cells (Calu-3) are used as relevant target cells to investigate their cytotoxicity. We observe a clear short-term loss of viability associated to reactive oxygen species generation, but with limited pro-inflammatory effects. On an actual cell-deposited mass-dose basis, the cytotoxicity of the nanosized fraction is similar to that of BWP, suggesting that the cytotoxicity is particle size independent. To conclude, brake wear dust contains substantial amount of metallic nanoparticles exhibiting toxicity for lung cells, and should warrant further consideration.

Hybrid Hydrophobin/Gold Nanoparticles: Synthesis and Characterization of New Synthetic Probes for Biological Applications

We report a simple and original method to synthesize gold nanoparticles in which a fungal protein, the hydrophobin Vmh2 from Pleurotus ostreatus, mixed to cetyltetrammonium bromide (CTAB) has been used as additional component in a one-step synthesis, leading to shell-like hybrid protein-metal nanoparticles (NPs). The nanoparticles have been characterized by ultra-violet/visible and infrared spectroscopies, and also by electron microscopy imaging. The results of these analytical techniques highlight nanometric sized, stable, hybrid complexes of about 10 nm, with a micelles-like hydrophobins rearrangement.