ABSTRACT
Biofilm formation triggered by uncontrolled protein adsorption, on medical devices is the leading cause of catheter-associated urinary tract infections (CAUTI) during implantation. Herein, we report a water-based, green and one-step strategy to functionalize surfaces of silicone catheters, poly(dimethylsiloxane) (PDMS), with antifouling and antimicrobial substances to avoid uncontrolled protein adsorption and microbial attachment. A novel synergetic formulation consisting of an anionic glycosaminoglycan (hyaluronic acid, HA) and a lysine-derived biocompatible cationic surfactant (Nε-myristoyl-lysine methyl ester, MKM) was prepared, resulting in the formation of nanoparticles (NPs, ca. 100-250 nm). Besides their high stability and long-lasting hydrophilicity in ambient and aqueous environments for 60 days, the nanometric layers (48 ± 3 nm) of HA-MKM NPs on PDMS showed no adsorption of BSA and lysozyme and substantially lower adsorption of fibrinogen as revealed by a quartz crystal microbalance with dissipation (QCM-D). In vitro antimicrobial test with S. aureus, E. coli, P. aeruginosa, P. mirabilis, C. albicans microbes under dynamic conditions revealed that the microbial growth was hampered by 85% compared with unmodified PDMS. Given the multiple functionalities, charges and diverse physiochemical properties of polysaccharide-lysine-based surfactant mixtures, this approach can be easily extended to the development of novel coatings on other silicone-based materials, thereby broadening potential applicability of PDMS-based biomaterials/devices in microfluidics, diagnostic biosensors and others.
ABSTRACT
Key biological functions involved in cell survival have been studied to understand the difference between the impact of exposure to TiO2 nanoparticles (TiO2-NPs) and their bulk counterparts (bulk-TiO2). By selecting a unicellular eukaryotic model organism and applying proteomic analysis an overview of the possible impact of exposure could be obtained. In this study, we investigated the early response of unicellular eukaryotic protozoan Tetrahymena thermophila exposed to TiO2-NPs or bulk-TiO2 particles at subtoxic concentrations for this organism. The proteomic analysis based on 2DE + nLC-ESI-MS/MS revealed 930 distinct protein spots, among which 77 were differentially expressed and 18 were unambiguously identified. We identified alterations in metabolic pathways, including lipid and fatty acid metabolism, purine metabolism and energetic metabolism, as well as salt stress and protein degradation. This proteomic study is consistent with our previous findings, where the early response of T. thermophila to subtoxic concentrations of TiO2 particles included alterations in lipid and fatty acid metabolism and ion regulation. The response to the lowest TiO2-NPs concentration differed significantly from the response to higher TiO2-NPs concentration and both bulk-TiO2 concentrations. Alterations on the physiological landscape were significant after exposure to both nano- and bulk-TiO2; however, no toxic effects were evidenced even at very high exposure concentrations. This study confirms the relevance of the alteration of the lipid profile and lipid metabolism in understanding the early impact of TiO2-NPs in eukaryotic cells, for example, phagocytosing cells like macrophages and ciliated cells in the respiratory epithelium.
Subject(s)
Nanoparticles/toxicity , Proteomics , Protozoan Proteins/metabolism , Tetrahymena thermophila/drug effects , Titanium/toxicity , Dose-Response Relationship, Drug , Lipid Peroxidation/drug effects , Metabolic Networks and Pathways/drug effects , Nanoparticles/chemistry , Particle Size , Protozoan Proteins/genetics , Tandem Mass Spectrometry , Tetrahymena thermophila/genetics , Tetrahymena thermophila/metabolism , Time Factors , Titanium/chemistryABSTRACT
Perindopril Erbumine (PER) is one of the newly used angiotensin-converting enzyme inhibitors (ACE inhibitors) and is used for the treatment of patients with hypertension and symptomatic heart failure. It has two main degradation pathways, i.e. the degradation by hydrolysis and the degradation by cyclization. An isothermal heat conduction microcalorimetry (MC) and high pressure liquid chromatography (HPLC) were used for the characterization of aqueous solutions of PER and its stability properties. The rates of heat evolved during degradation of perindopril were measured by MC as a function of temperature and pH and from these data rate constant and change in enthalpy of the reactions were determined. With the HPLC method the concentration of perindopril and its degradation products were measured as a function of time in aqueous solutions of different pH that were stored at different temperatures. We demonstrated that reactions of degradation of perindopril at observed conditions follow the first order kinetics. The Arrhenius equation for each pH was determined. At pH 6.8 only one degradation pathway is present, i.e. the degradation by hydrolysis. Degradation constants for this pathway calculated from MC data are in good agreement with those obtained from HPLC. MC as a non-specific technique was shown to be useful in studies of PER when one reaction was present in the sample and also when more chemical and physical processes were simultaneously running.
Subject(s)
Angiotensin-Converting Enzyme Inhibitors/chemistry , Perindopril/chemistry , Thermodynamics , Calorimetry/methods , Chemistry, Pharmaceutical , Chromatography, High Pressure Liquid/methods , Drug Stability , Drug Storage , Hydrogen-Ion Concentration , Kinetics , Pharmaceutical SolutionsABSTRACT
The stability properties of enalapril maleate (EM) and of different tablet formulations including EM were studied by isothermal microcalorimetry and by high performance liquid chromatography (HPLC). It was shown that water content of the sample and elevated temperature have a high impact on stability properties of the substance itself and of the formulations including this substance. The degradation is more extensive at higher water content and at elevated temperature. The type of the tablet formulation (5 or 20mg EM tablet formulation) also has an impact: the 5 EM tablet formulation is the less stable one. The heat output of individual tablet formulations was used to evaluate the enthalpy changes and to calculate the difference in the amount of degraded EM between various samples. These results agreed satisfactorily with those obtained by HPLC. Isothermal microcalorimetry proved to be a fast and predictive method that could be used in preformulation studies to accelerate the pharmaceutical development and shorten the time before launching the product to the market.