Small changes in PDMMLA structure influence the adsorption behavior of ECM proteins and syndecan-4 on PDMMLA derivative surfaces: Experimental validation by tensiometric surface force measurements.

Cardiovascular diseases are the leading cause of death around the world according to the World Health Organization. In-stent restenosis is an inflammatory response of the immune system to endovascular stent implantation in atherosclerotic patients. Biocompatible and biodegradable polymers are of great interest in this field in order to limit the side effects of stent treatments. Poly([R,S]-3,3-dimethylmalic acid) (PDMMLA) is a new biodegradable statistical polyester which presents promising properties as a stent coating. In this work, we studied by dynamic tensiometry, the adhesion of extracellular matrix proteins (bovine serum albumin, fibronectin, fibrinogen, and vitronectin) and plasma membrane proteoglycan (syndecan-4) on three PDMMLA derivatives with different hydrophilicity levels. The results show that proteins have different adhesion profiles and affinity on these surfaces. They show similar behavior on the most hydrophilic surface, making hydrophilic, ionic and hydrogen type bonds. Then we compared each protein's individual profile to that of a mixture of all studied proteins. The comparison shows that vitronectin and syndecan-4 are the quantitatively dominating proteins adsorbed by specific interactions. Based on the results from previous studies, this work allowed us to identify the most important PDMMLA surface as a promising biomaterial for bioactive stent-coating.

A facile route to glycated albumin detection.

In this paper we propose an easy way to detect the glycated form of human serum albumin which is biomarker for several diseases such as diabetes and Alzheimer. The detection platform is a label free impedimetric immunosensor, in which we used a monoclonal human serum albumin antibody as a bioreceptor and electrochemical impedance as a transducing method. The antibody was deposited onto a gold surface by simple physisorption technique. Bovine serum albumin was used as a blocking agent for non-specific binding interactions. Cyclic voltammetry and electrochemical impedance spectroscopy were used for the characterization of each layer. Human serum albumin was glycated at different levels with several concentrations of glucose ranging from 0 mM to 500 mM representing physiological, pathological (diabetic albumin) and suprapathological concentration of glucose. Through the calibration curves, we could clearly distinguish between two different areas related to physiological and pathological albumin glycation levels. The immunosensor displayed a linear range from 7.49% to 15.79% of glycated albumin to total albumin with a good sensitivity. Surface plasmon resonance imaging was also used to characterize the developed immunosensor.

Electrochemical Impedance Spectroscopy on Interdigitated Gold Microelectrodes for Glycosylated Human Serum Albumin Characterization.

Glycosylated albumin is considered as a potentially accurate indicator of shorter-term average glucose concentration compared with the current standard HbA1c and as such, it is attracting the interest of the scientific community as a possible diagnosis marker for diabetic patients. The purpose of this paper is to achieve a better understanding of the glycation effect of albumin on its electrochemical properties. That is done through the use of Interdigitated gold microelectrodes (IDGE) as support in a label free impedimetric immunosensor for the detection of human serum albumin detection in glycated (GA) and non-glycated (HSA) form. Anti-human serum albumin, a monoclonal antibody, was physisorbed on the surface of IDGE and used as a HSA/GA bioreceptor. Electrochemical impedance spectroscopy, cyclic voltammetry, and surface plasmon resonance imaging (SPRi) were used for the characterization of the grafted layers onto the gold surface. A detection range from 1 to 401 ng/mL of non glycated HSA antigen in phosphate buffered saline buffer was obtained with the impedance spectroscopy technique. The experiment led to the observation of a significant impedance difference between the glycated and non-glycated antigen of HSA. SPRi measurements confirmed these findings and allowed us to suggest an increase of the dielectric permittivity for human serum albumin upon glycation.

Impact of chlorhexidine digluconate and temperature on curli production in Escherichia coli-consequence on its adhesion ability.

Chlorhexidine-Digluconate (CHX-Dg) is a biocide widely used as disinfectant or antiseptic in clinical and domestic fields. It is often found in the formulation of solutions to treat superficial wounds. Nevertheless, few studies have focused on its effects on Escherichia coli while this bacterium is commonly involved in mixed infections. Therefore, the impact of CHX-Dg and temperature on E. coli was investigated; particularly the curli production. In accordance with bibliographic data, the curli production decreased when the temperature of the culture was shift from 30 °C to 37 °C. The bacterial adhesion to abiotic surfaces was also reduced. Surprisingly, the curli production at 37 °C was maintained in presence of antiseptic and the bacterial adhesion was improved at a very low concentration (1 µg ml-1) of CHX-Dg. Complementary investigations with a cpxR mutant demonstrated that the CpxA/R-TCS (Two-Component System) is involved in the temperature-dependent control of the curli expression. Indeed, the curli production was not altered by the growth temperature in the mutant. Otherwise, no relationship between CHX-Dg and the Cpx-TCS was shown. A subsequent proteomic investigation revealed the alteration of the production of 44 periplasmic and outer membrane proteins in presence of CHX-Dg. These proteins are involved in the transport of small molecules, the envelope integrity, the stress response as well as the protein folding.

Dynamic contact angle analysis of protein adsorption on polysaccharide multilayer's films for biomaterial reendothelialization.

Atherosclerosis is a major cardiovascular disease. One of the side effects is restenosis. The aim of this work was to study the coating of stents by dextran derivates based polyelectrolyte's multilayer (PEM) films in order to increase endothelialization of injured arterial wall after stent implantation. Films were composed with diethylaminoethyl dextran (DEAE) as polycation and dextran sulphate (DS) as polyanion. One film was composed with 4 bilayers of (DEAE-DS)4 and was labeled D-. The other film was the same as D- but with an added terminal layer of DEAE polycation: (DEAE-DS)4-DEAE (labeled D+). The dynamic adsorption/desorption of proteins on the films were characterized by dynamic contact angle (DCA) and atomic force microscopy (AFM). Human endothelial cell (HUVEC) adhesion and proliferation were quantified and correlated to protein adsorption analyzed by DCA for fibronectin, vitronectin, and bovine serum albumin (BSA). Our results showed that the endothelial cell response was optimal for films composed of DS as external layer. Fibronectin was found to be the only protein to exhibit a reversible change in conformation after desorption test. This behavior was only observed for (DEAE-DS)4 films. (DEAE-DS)4 films could enhance HUVEC proliferation in agreement with fibronectin ability to easily change from conformation.

Supported protein G on gold electrode: characterization and immunosensor application.

In this work, we study the electrochemical properties of protein layer grafted on gold electrode for C-reactive protein detection. Two CRP-antibody immobilization methods were used: the first method is based on direct physisorption of CRP-antibody onto the gold surface and the second method is based on oriented CRP-antibody with protein G intermediate layer. The two developed immunosensors were tested against CRP antigen in phosphate buffer saline solution and in human plasma. The electrochemical characterization of each immobilized layers was achieved by cyclic voltammetry and impedance spectroscopy. The morphology of the deposited biomolecules was observed by Atomic Force Microscopy and the roughness was measured. Moreover, contact angle measurement was used for wettability studies. The response of the developed immunosensors was reproducible, rapid, and highly stable and a detection limit of 100 fg/mL and 10 pg/mL antigen was observed with and without protein G respectively. The developed immunosensors was used for CRP detection in human plasma.

PEM anchorage on titanium using catechol grafting.

BACKGROUND: This study deals with the anchorage of polyelectrolyte films onto titanium surfaces via a cathecol-based linker for biomedical applications. METHODOLOGY: The following study uses a molecule functionalized with a catechol and a carboxylic acid: 3-(3,4-dihydroxyphenyl)propanoic acid. This molecule is anchored to the TiO(2) substrate via the catechol while the carboxylic acid reacts with polymers bearing amine groups. By providing a film anchorage of chemisorption type, it makes possible to deposit polyelectrolytes on the surface of titanium. PRINCIPAL FINDINGS: Infrared spectroscopy (ATR-FTIR), X-ray photoelectron spectroscopy (XPS), contact angle and atomic force microscopy (AFM) measurements show that the different steps of grafting have been successfully performed. CONCLUSIONS: This method based on catechol anchorage of polyelectrolytes open a window towards large possibilities of clinical applications.

Assessing bacterial adhesion using DLVO and XDLVO theories and the jet impingement technique.

In this study, the adhesion of two bacterial strains (Pseudomonas stutzeri PS, and Staphylococcus epidermidis, SE) to the glass and the indium tin oxide (ITO)-coated glass surfaces was examined qualitatively and quantitatively using the theoretical approaches and the jet impingement technique. A comparison between the DLVO and the extended DLVO (XDLVO) theories showed that the XDLVO predictions of bacterial adhesion and its reversibility are more accurate than DLVO predictions. The adhesion tests revealed that PS bacteria has much better adhesion rate than SE bacteria to both material surfaces, as predicted by XDLVO approach. Also both bacterial strains adhered better to the hydrophobic ITO-coated glass than to the hydrophilic glass surface, as predicted theoretically. Moreover, the microjet impingement technique was used not only to assess the bacterial adhesion strength on both materials, but also to verify the adhesion reversibility. The detachment stress values demonstrated that PS bacterial cells adhered strongly and irreversibly in the primary energy minimum, while SE bacterial cells adhered weakly and reversibly in the secondary energy minimum on both substrata surfaces. Also, the adhesion of both bacterial strains was found better and stronger on the hydrophobic ITO-coated glass surface than on the hydrophilic glass surface.