Lionel Marcon

Cellular and in vivo toxicity of functionalized nanodiamond in Xenopus embryos

Recently, nanodiamond particles (ND) have emerged as a promising tool in the field of nanobiotechnology. However, studies about the impact of ND on living organisms are still limited to raw materials and primarily confined to in vitro studies. In this work, we investigated the cytotoxicity and in vivo toxicity of ND correlated with their chemical surface functionality (-OH, -NH2 or -CO2H). Two model systems have been used, human embryonic kidney 293 (HEK293) cells and Xenopus laevis embryos. Cell viability assays showed that ND were not cytotoxic to HEK293 cells for concentrations below 50 μg mL−1. Our data suggest that the cytotoxicity may be due to the affinity of cationic particles for the negatively charged cell membrane. In parallel, visual monitoring of microinjected early-stage embryos showed a potential embryotoxicity and teratogenicity for carboxylated ND-CO2H. ND seem to have a negative impact on the gastrulation and neurulation stages inducing phenotypical abnormalities and high mortality.

Functionalization of Diamond Nanoparticles Using “Click” Chemistry

The paper reports on covalent linking of different alkyne-containing (decyne, ethynylferrocene, and N-propargyl-1-pyrenecarboxamide) compounds to azide-terminated nanodiamond (ND) particles. Azide-terminated particles (ND-N(3)) were obtained from amine-terminated nanodiamond particles (ND-NH(2)) through the reaction with 4-azidobenzoic acid in the presence of a carbodiimide coupling agent. Functionalized ND particles with long alkyl chain groups can be easily dispersed in various organic solvents without any apparent precipitation after several hours. The course of the reaction was followed using Fourier transform infrared (FT-IR) spectroscopy, UV/vis spectroscopy, fluorescence, cyclic voltammetry, thermogravimetric analysis (TGA), and particle size measurements. The surface loading of pyrene bearing a terminal acetylene group was found to be 0.54 mmol/g. Because of its gentle nature and specificity, the chemistry developed in this work can be used as a general platform for the preparation of functional nanoparticles for various applications.

Combined nanogap nanoparticles nanosensor for electrical detection of biomolecular interactions between polypeptides

A concept for the electrical detection of a biological interaction is proposed, mainly based on the conductance variation of a nanometer size-gap (typically less than 100 nm) between two planar electrodes. A functionalized surface was used in the vicinity of the gap in order to concentrate the ligand/receptor complex between the electrodes. The chemistry chosen for the immobilization of the ligand on the biosensor surface is compatible with peptide structures. The receptor in solution was labeled with gold particles which can be inserted into the gap. A significant conductance variation was observed without having to use a silver enhancer solution in the case of biotin/streptavidin or biotin/antibiotin antibodies model ligand/receptor interactions.

The antimicrobial effect of silicon nanowires decorated with silver and copper nanoparticles.

The paper reports on the preparation and antibacterial activity of silicon nanowire (SiNW) substrates coated with Ag or Cu nanoparticles (NPs) against Escherichia coli (E. coli) bacteria. The substrates are easily prepared using the metal-assisted chemical etching of crystalline silicon in hydrofluoric acid/silver nitrate (HF/AgNO3) aqueous solution. Decoration of the SiNWs with metal NPs is achieved by simple immersion in HF aqueous solutions containing silver or copper salts. The SiNWs coated with Ag NPs are biocompatible with human lung adenocarcinoma epithelial cell line A549 while possessing strong antibacterial properties to E. coli. In contrast, the SiNWs decorated with Cu NPs showed higher cytotoxicity and slightly lower antibacterial activity. Moreover, it was also observed that leakage of sugars and proteins from the cell wall of E. coli in interaction with SiNWs decorated with Ag NPs is higher compared to SiNWs modified with Cu NPs.

Photochemical Immobilization of Proteins and Peptides on Benzophenone-Terminated Boron-Doped Diamond Surfaces

The successful covalent linking of green fluorescence protein and streptavidin to patterned benzophenone-modified boron-doped diamond (BDD) electrodes is demonstrated. Photoreactive benzophenone moieties were covalently grafted to oxidized diamond surfaces via an esterification reaction. Patterned BDD surfaces were obtained using a UV/ozone lithographic approach either on hydrogen-terminated BDD or on poly(ethylene)-glycol-modified BDD surfaces. UV light (lambda = 365 nm) irradiation of the patterned BDD surfaces in the presence of green fluorescence protein (GFP) or streptavidin resulted in the covalent immobilization of the proteins. The presence of poly(ethylene) glycol chains reduces significantly the nonspecific adsorption of the proteins. The success of the photoimmobilization of streptavidin was evidenced through biomolecular interaction with avidin. The preservation of the biological activity was furthermore underlined by photoimmobilization of peptides directly onto benzophenone modified BDD using a photomask.

Cell micropatterning on superhydrophobic diamond nanowires

Cell micropatterning was achieved in a spatially controlled manner based on heterogeneously wetted superhydrophilic/superhydrophobic diamond nanowire (NW) surfaces. Diamond NWs were synthesized on boron-doped diamond substrates using reactive ion etching and functionalized with octadecyltrichlorosilane to achieve superhydrophobicity. Superhydrophilic motifs of 400×400 μm(2) and 10×10 μm(2) single cell-sized motifs, surrounded by superhydrophobic regions, were then generated by selectively exposing the substrates to UV light. This design allowed successful patterning of single HeLa and MCF-10A cells within the superhydrophilic regions without additional surface modification. To add a further level of complexity, micropatterned co-cultures were obtained using bovine serum albumin to promote cell adhesion. This method is simple and does not require any complicated processing steps such as mask deposition or template removal. Potential applications are in the development of cell-based biological assays in well-controlled and biologically relevant environments.

Cell adhesion properties on chemically micropatterned boron-doped diamond surfaces.

The adhesion properties of living cells were investigated on a range of chemically modified boron-doped diamond (BDD) surfaces. We studied the influence of oxidized, H-, amine- (NH(2)-), methyl- (CH(3)-), trifluoromethyl- (CF(3)-) and vinyl- (CH(2)═CH-) terminated BDD surfaces on human osteosarcoma U2OS and mouse fibroblast L929 cells behavior. Cell-surface interactions were analyzed by fluorescence microscopy in terms of cell attachment, spreading and proliferation. U2OS cells poorly adhered on hydrophobic surfaces and their growth was blocked. In contrast, L929 cells were mainly influenced by the presence of perfluoroalkyl chains in regard to their morphology. The results were subsequently applied to selectively micropattern U2OS cells on dual hydrophobic/hydrophilic surfaces prepared by a UV/ozone lithographic approach. U2OS cells colonized preferentially hydrophilic (oxide-terminated) motifs, forming confluent arrays with distinguishable edges separating the alkyl regions.

Characterization of nanogap chemical reactivity using peptide-capped gold nanoparticles and electrical detection.

Nanogaps are usually combined with synthetic or biological molecules to produce nanodevices having novel properties. This combination is better realized by controlling the chemical properties of the nanogap. We show here that the presence of a specific chemical group inside nanogaps (30-90 nm) can be probed electrically using 10 nm gold nanoparticles derivatized by complementary functional groups. 100-10(4)-fold current increases were observed following the site-specific insertion of gold nanoparticles into the nanogap.

Biocompatibility of semiconducting silicon nanowires

Abstract: Silicon nanowires (SiNW) have been shown to be potential candidates for biological applications such as drug delivery systems, in vivo imaging agents and biosensors. However, concerns have been raised over adverse effects that SiNW may exert on biological systems. The first objective of this chapter is to offer a general review of the studies on the biocompatibility of SiNW in vitro and in vivo . The second objective is then to discuss the relevance of published data and present trends in the toxicity studies of SiNW in relation to their possible future applications.

Fundamental studies in nanosciences at the Institute of Electronics, Microelectronics, and Nanotechnology (IEMN)

This paper gives an overview over the fundamental research in nanosciences at the Institute of Electronics, Microelectronics and Nanotechnology (IEMN). We present some highlights from the numerical simulation of the electronic structure of nanowires and nanotubes, the charge spectroscopy of Si nanoparticles and C nanotubes, the scanning tunnelling spectroscopy of semiconductor quantum dots, to research in surface science for bio-screening.