Proteins dominate in the surface layers formed on materials exposed to extracellular polymeric substances from bacterial cultures.

The chemical compositions of the surface conditioning layers formed by different types of solutions (from isolated EPS to whole culture media), involving different bacterial strains relevant for biocorrosion were compared, as they may influence the initial step in biofilm formation. Different substrata (polystyrene, glass, steel) were conditioned and analyzed by X-ray photoelectron spectroscopy. Peak decomposition and assignment were validated by correlations between independent spectral data and the ubiquitous presence of organic contaminants on inorganic substrata was taken into account. Proteins or peptides were found to be a major constituent of all conditioning layers and polysaccharides were not present in appreciable concentrations; the proportion of nitrogen which may be due to DNA was lower than 15%. There was no significant difference between the compositions of the adlayers formed from different conditioning solutions, except for the adlayers produced with tightly bound EPS extracted from D. alaskensis.

Oxidation of laccase for improved cathode biofuel cell performances.

Graphite rods were modified by substituted aryldiazonium salts allowing subsequent laccase immobilisation and direct electron transfer at the cathode. Two covalent enzyme immobilisation methods were performed with carboxy and amino substituted grafted groups, either via the formation of an amide bond or a Schiff base between the glycosidic groups of the enzyme and the amino groups on the electrode surface, respectively. Laccase adsorption efficiency was consistently compared to the covalent attachment method on the same carbon surface, showing that the latter method led to a higher immobilisation yield when the electrode surface was functionalised with carboxylic groups, as shown from both laccase activity measurement towards an organic reducing substrate, ABTS, and quantitative XPS analysis. Both analytical methods led to similar laccase surface coverage estimations. From activity measurements, when laccase was covalently immobilised on the electrode functionalised with carboxylic groups, the surface coverage was found to be 43 ± 2% whereas it was only 10 ± 3% when laccase was adsorbed. Biocatalysed dioxygen reduction current was also higher in the case of covalent immobilisation. For the first time, oxidised laccase performances were compared to unmodified laccase, showing significant improved efficiency when using oxidised laccase: the current obtained with oxidised laccase was 141 ± 37 µA cm(-2) compared to 28 ± 6 µA cm(-2) for unmodified laccase after covalent immobilisation of the enzyme on a graphite electrode functionalised with carboxylic groups.

Chitosan-coated electrospun nanofibers with antibacterial activity.

Charged nanofibers were prepared by electrospinning (ESP) poly(ε-caprolactone) with a copolymer bearing carboxylic acid functions. The presence of these functions allowed exposing some negative charges on the fiber surface, by dipping the fibers in a phosphate buffer. A layer of chitosan, a polycation in acidic medium, was then deposited on the nanofiber surface, thanks to electrostatic attraction. Fibers were characterized at each step of the process and the influence of the copolymer architecture on chitosan deposition was discussed. The antibacterial activity of the resulting fibers was finally assessed.

Conditioning materials with biomacromolecules: composition of the adlayer and influence on cleanability.

The influence of substrate hydrophobicity and biomacromolecules (dextran, bovine serum albumin - BSA) adsorption on the cleanability of surfaces soiled by spraying aqueous suspensions of quartz particles (10-30µm size), then dried, was investigated using glass and polystyrene as substrates. The cleanability was evaluated using radial flow cell (RFC). The surface composition was determined by X-ray photoelectron spectroscopy (XPS). The interpretation of XPS data allowed the complexity due to the ubiquitous presence of organic contaminants to be coped with, and the surface composition to be expressed in terms of both the amount of adlayer and the mass concentration of adlayer constituents. When soiled with a suspension of particles in water, glass was much less cleanable than polystyrene, which was attributed to its much lower water contact angle, in agreement with previous observations on starch soil. Dextran was easily desorbed and did not affect the cleanability. The presence of BSA at the interface strongly improved the cleanability of glass while the contact angle did not change appreciably. In contrast, soiling polystyrene with quartz particles suspended in a BSA solution instead of water did not change markedly the cleanability, while the contact angle was much lower and the aggregates of soiling particles were more flat. These observations are explained by the major role of capillary forces developed upon drying, which influence the closeness of the contact between the soiling particles and the substrate and, thereby, the adherence of particles. The capillary forces are proportional to the liquid surface tension and depend in a more complex way on contact angles of the particles and of the substrate. The dependence of cleanability on capillary forces, and in particular on the liquid surface tension, is predominant as compared with its dependence on the size and shape of the soiling aggregates, which influence the efficiency of shear forces exerted by the flowing water upon cleaning.

Calibration of the X-ray photoelectron spectroscopy binding energy scale for the characterization of heterogeneous catalysts: is everything really under control?

Investigations of X-ray photoelectron spectra from solid samples need corrections for the surface charging effect. For powder samples such as heterogeneous catalysts and their supports, the C-(C,H) component of the C 1s peak is often used as an internal standard for the calibration of the binding energy scale. Although this method is widely recognized as suitable for the study of heterogeneous catalysts, we show that a significant calibration bias can be encountered upon comparing samples with different bulk composition. In this paper, a series of SiO2-Al2O3 supports and Pd/SiO2-Al2O3 catalysts with various Si/Al ratios were studied. The spectra issued from these samples were processed with the classical calibration method on the basis of the carbon peak. Important discrepancies in the relative position of the photoelectron peaks were noticed. After systematically discarding instrument-related issues, a true chemical influence of the bulk matrix on the analyzed surface species was evidenced. The extent of this chemical effect was dependent on the composition of the sample and more precisely on its ionicity. Two possible mechanisms for this chemical effect were proposed and discussed. Finally, an alternative calibration method was offered.

Surface characterization, collagen adsorption and cell behaviour on poly(L-lactide-co-glycolide).

Poly(L-lactide-co-glycolide) (PLG) was modified through the adsorption of collagen to improve the behaviour of fibroblasts and osteoblasts. As reference materials cell-resistant polystyrene (PS) and cell-conductive tissue-culture polystyrene (TCPS) were also evaluated. The physicochemical surface properties of the materials were studied by X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), and water contact angle measurements. The morphology of cells was examined using optical microscopy, while their growth was evaluated by both crystal violet and MTT tests. Nitric oxide level and protein concentration were tested in cell supernatants. The results showed that the adsorbed amount and the organization of the adsorbed collagen were influenced by surface hydrophobicity. Cell culture experiments on native substrates revealed that cell attachment, spreading and growth enhanced, depending on the substrate, in the following order: PS

Surface chemical composition of diatoms.

Among diatoms, Phaeodactylum tricornutum is a peculiar species that exists in three morphotypes with distinct cell wall structures and low silica content. X-ray photoelectron spectroscopy (XPS) analysis was performed on P. tricornutum and compared with diatom Thalassiosira pseudonana; the results provide new information on the chemical composition (elements, chemical functions, classes of biochemical compounds) of the cell surface. Two types of silicon were found: condensed silica (SiO(2)) and weakly polymerised silicate. Cells of T. pseudonana showed the highest concentration of silicon, with a majority in the form of condensed silica. For the fusiform and triradiate morphotypes of P. tricornutum, the majority of the small concentration of silica found was in the form of weakly polymerised silicate. For all morphotypes of P. tricornutum, higher polysaccharide concentrations replaced silica as a structural part of the cell wall. In both diatoms, a high concentration of lipids was measured, in the form of carboxylic esters. Protonated nitrogen and phosphate were found in correlated amounts and attributed not only to phospholipids but also to phosphoproteins. Chloride ions characterised by a high electron density might be associated to these moieties. Sulfate groups were also detected, principally in P. tricornutum, and attributed to monoesters of polysaccharides.

Principal component analysis: a versatile method for processing and investigation of XPS spectra.

Given the relevance of principal component analysis (PCA) to the treatment of spectrometric data, we have evaluated potentialities and limitations of such useful statistical approach for the harvesting of information in large sets of X-ray photoelectron spectroscopy (XPS) spectra. Examples allowed highlighting the contribution of PCA to data treatment by comparing the results of this data analysis with those obtained by the usual XPS quantification methods. PCA was shown to improve the identification of chemical shifts of interest and to reveal correlations between peak components. First attempts to use the method led to poor results, which showed mainly the distance between series of samples analyzed at different moments. To weaken the effect of variations of minor interest, a data normalization strategy was developed and tested. A second issue was encountered with spectra suffering of an even slightly inaccurate binding energy scale correction. Indeed, minor shifts of energy channels lead to the PCA being performed on incorrect variables and consequently to misleading information. In order to improve the energy scale correction and to speed up this step of data pretreatment, a data processing method based on PCA was used. Finally, the overlap of different sources of variation was studied. Since the intensity of a given energy channel consists of electrons from several origins, having suffered inelastic collisions (background) or not (peaks), the PCA approach cannot compare them separately, which may lead to confusion or loss of information. By extracting the peaks from the background and considering them as new variables, the effect of the elemental composition could be taken into account in the case of spectra with very different backgrounds. In conclusion, PCA is a very useful diagnostic tool for the interpretation of XPS spectra, but it requires a careful and appropriate data pretreatment.