Synthesis of polymer materials for use as cell culture substrates

Up to today, several techniques have been used to maintain cells in culture for studying many aspects of cell biology and physiology. More often, cell culture is dependent on proper anchorage of cells to the growth surface. Thus, poly-l-lysine, fibronectin or laminin are the most commonly used substrates. In this study, electrosynthesized biocompatible polymer films are proposed as an alternative to these standard substrates. The electrosynthesized polymers tested were polyethylenimine, polypropylenimine and polypyrrole. Then, the adhesion, proliferation and morphology of rat neuronal cell lines were investigated on these polymer substrates in an attempt to develop new and efficient polymer materials for cell culture. During their growth on the polymers, the evolution of the cell morphology was monitored using both confocal microscopy and immunohistochemistry, leading to the conclusion of a normal development. An estimation of the adhesion and proliferation rates of rat neuronal cell cultures indicated that polyethylenimine and polypropylenimine were the best substrates for culturing olfactory neuronal cells. A method to favour the differentiation of the neuronal cells was also developed since the final aim of this work is to develop a biosensor for odour detection using differentiated neuronal cells as transducers. Consequently, a biosensor was microfabricated using silicon technology. This microsystem allowed us to culture the cells on a silicon wafer and to position the cells on certain parts of the silicon wafer.     

Electrochemistry at capped platinum nanoparticle Langmuir Blodgett films: A study of the influence of platinum amount and of number of LB layers

Platinum nanoparticles (n-Pt), over-grafted with 2-thiophenecarbonyl chloride are assembled on gold electrodes, by the Langmuir Blodgett (LB) technique using behenic acid (BHA) as promoting agent. These layers are electrochemically active without any preliminary activation. The [Fe(CN)₆]^3−/4− redox couple was used as electrochemical probe. This paper reports on the influence of the number of deposited LB layers, and the n-Pt density on the electrochemical response. n-Pt density was modified by the change of the “BHA/n-Pt” ratio. Cyclic voltammograms of “[Fe(CN)₆]^3−/4−” were observed whatever the coating conditions. As soon as the first layer was deposited the electrochemical response was associated to the n-Pt coverage, its response slightly increased up to a steady state for five or seven layers. As expected, the increase of the Pt density favored the increase of the current density. XPS analysis performed before and after electrochemical cycling showed that 4-mercaptoaniline capped platinum nanoparticles, and their over grafting were chemically and electrochemically stable. Analysis of influence of the number or the n-Pt density of the layers showed that the electrochemically active part of LB electrodes was provided by the last layer plus a part of the underlying one.     

Electronic transfer through Langmuir-Blodgett layers of capped platinum nanoparticles: An electrochemical approach

Amine functionalized platinum nanoparticles have been modified by over-grafting two different molecules, 2-thiophenecarbonyl chloride (Pt-1) and 1-hexyl-4-(4-isothiocyanatophenyl)-bicyclo (2, 2, 2) octane (Pt-2). Cyclic voltammetry was performed at gold electrodes coated with Langmuir-Blodgett (LB) mixed films of Pt-1 and Pt-2 nanoparticles, and behenic acid. From five layers the electrochemical response was essentially provided by the last LB component. The electrochemical responses towards the [Fe(CN)₆]^3−/4− couple were strongly influenced by the nature of the over-grafted molecules: films of Pt-2 presented an almost complete blocking effect, while films of Pt-1 allowed the redox reaction to occur on Pt nanoparticles. In order to understand the reasons for such different behaviors we built up hetero-nanostructures by superposing Pt-1 and Pt-2 LB layers in different ways, yielding different kinds of “sandwich” structures. The electrochemical response depended on the electrode ending. When Pt-1 nanoparticles were in the outer layer, in contact with the electrolyte solution, the electrode was electroactive toward the redox probe, while when Pt-2 layers were in the outer layer no electroactivity was detected. For sandwiches made of Pt-1, with a variable thickness of an intercalated film of Pt-2, the electrode response to [Fe(CN)₆]^3−/4− was modulated by the thickness of the inter-layer: the thicker the layer, the lower the response.     

Shear effect on dynamic behavior of microcantilever beam with manufacturing process defects

This paper is concerned with the investigation of the shear effect on the dynamic behavior of a thin microcantilever beam with manufacturing process defects. Unlike the Rayleigh beam model (RBM), the Timoshenko beam model (TBM) takes in consideration the shear effect on the resonance frequency. This effect become significant for thin microcantilever beams with larger slenderness ratios that are normally encountered in MEMS devices such as sensors. The TBM model is presented and analyzed by numerical simulation using Finite Element Method (FEM) to determine corrective factors for the correction of the effect of manufacturing process defects like the underetching at the clamped end of the microbeam and the nonrectangular cross section of the area. A semi-analytical approach is proposed for the extraction of the Young’s modulus from 3D FEM simulation with COMSOL Multiphysics software. This model was tested on measurements of a thin chromium microcantilever beam of dimensions (80 × 2 × 0.95 μm³). Final results indicate that the correction of the effect of manufacturing process defects is significant where the corrected value of Young’s modulus is very close to the experimental results and it is about 280.81 GPa.

     

Photoelectrochemical characterization of the p-Cu2O-non aqueous electrolyte junction

The current-voltage characteristics of the p-Cu₂O electrolyte junctions were analyzed. The electrode decomposition process was defined in the dark and under illumination in the liquid ammoniate of sodium iodide (LASI). It was shown that the photodecomposition process is slower in LASI. The photocurrents at the p-Cu₂O/LASI junctions were described using the Gärtner model. Some typical solid parameters such as energy gap and minority carrier diffusion length were determined.     

Surface modification of p-Si by a polyethylenimine coating: influence of the surface pre-treatment. Application to a potentiometric transducer as pH sensor

p-Si electrodes coated with linear polyethylenimine (L-PEI) allow the fabrication of a pH sensitive film for potentiometric transducers. The coating is realized in one step through the anodic oxidation of pure ethylenediamine (EDA) charged with 0.1 M LiCF₃SO₃ (Lithium Triflate). Such an electrochemical procedure leads to the thickness control of the coating. The best silicon surface pre-treatment before any coating is obtained with potassium dichromate in sulfuric acid, which leads to OH-terminated p-Si. This pre-treatment allows a uniform thin coating. In this work, the thickness is 2.6 nm. The pH response is high and close to 50 mV per pH unit.     

Electrodeposition and characterization of silane thin films from 3-(aminopropyl)triethoxysilane

3-(aminopropyl)triethoxysilane based films have been electrodeposited directly on polycrystalline gold and gold (111) electrodes in aqueous 3-(aminopropyl)triethoxysilane based electrolyte and in tetrahydrofurane based electrolyte. These films were characterized by means of IR-ATR and X-ray photoelectron spectroscopies. The film morphology was investigated by scanning tunneling microscopy while the film growth was observed by ellipsometry measurements. The vibrationnal and X-ray photoelectron analysis suggest that the chemical composition of the electrodeposited films either in liquid tetrahydrofurane or in liquid 3-(aminopropyl)triethoxysilane is identical. The resulting coating thickness is different for the same biasing time in the two liquid media. The gold surface is coated irreversibly by an amino terminated film of great interest for sensor applications which was used as the functionalized part of a surface plasmon resonance biosensor to monitor α-lactalbumin graft.     

Electrocatalyzed synthesis of polypeptides on platinum surface in concentrated glycine electrolytes and ab initio calculations coupled to spectroscopic analysis

The anodic oxidation of glycine in aqueous electrolyte on smooth platinum electrode was carried out by electrochemical quartz crystal microbalance coupled to cyclic voltammetry technique. Here, we performed electrochemical experiments in concentrated glycine-based electrolyte. Contrary to studies made in diluted medium, the reaction we describe leads to a strongly grafted polymer on the surface at pH values superior to 6, and up to 13, in an irreversible way. The electrodeposited mass is even very significant at pH 13. Several methods such as AFM topography and spectroscopic techniques were performed to characterize the resulting coating. The polymer growth in alkaline conditions during the anodic oxidation of glycine in water probably involves an electrocatalytic step. We showed the presence of amide bonds and then polypeptide formation on the smooth platinum surface. Periodic ab initio calculations on polyglycine II were performed and compared to XPS and vibrational spectra.     

The pH response of the InP/liquid ammonia interface at 223 K: A pure nernstian behavior

Liquid ammonia (NH₃liq.) provides an original electrochemical environment onto semiconductors electrodes (SC). Its particular interest is that water influences can be neglected in opposition to a lot of non-aqueous other solvents. Fundamentals electrochemistries of the SC are related to the energy diagram of the interface. In this paper it is established into NH₃liq., on both types, on p- and n-InP by flat band potential (V_fb) measurements. The V_fb are determined over the whole range of pH that reaches 33 pH units in this non-aqueous solvent. InP exhibits a pure nernstian behavior with a specific 44 mV/pH slope at 223 K. This is particular compare to the “under nernstian” dependency, observed in water onto InP. The actual band positions are related to the initial chemical state of the InP surface, since oxide free surfaces and thin native oxide covered surfaces differ slightly. Reproducible contrasted results from the acid-base equilibrium, which is supported by the V_fb according to the pH, for singular interfacial chemistry. This aspect is confirmed by XPS measurements performed before and after InP immersion into NH₃liq. They establish the perfect stability of the initial chemical composition of the semiconducting surface in contact with NH₃liq. whatever its pH conditions. NH₃liq. appears as an inert solvent which is able to create an acid-base equilibrium onto InP surfaces. Each InP surface chemistry support its own linear V_fbvs. pH variation. In NH₃liq., the poor water control on the building of the Helmholtz layer is well shown by the perfect V_fb alignment position from the intermediated pH buffered solution obtained from the addition of tetraethyl ammonium hydroxide ((Et)₄N⁺,OH⁻) dissolved in water (20%).     

Influence of water addition on structural and electrochemical behaviour of liquid ammoniates: A study of LiClO4·4NH 3 ammoniate/H2O mixtures

The ammoniates of lithium perchlorate (LiClO₄•xNH₃) can be used as liquid or solid electrolytes in lithium batteries. The compounds possess a very low ammonia pressure and a large electroactivity range at room temperature. The presence of water in the electrolyte is one of the main problems for the lithium anode. From calorimetric measurements on the water-ammoniate system and an X-ray powder diffraction study on crystalline compounds that appear on lithium dipped in the electrolyte, a maximum water content, x = 0.2, can be derived. Above this limit, the thermodynamic properties of the compounds change and the lithium reacts dramatically with the electrolyte. Electrochemical stability ranges behave, over all the water molar fractions studied, in accordance with structural features.