Formulation and Structural Study of a Biocompatible Water-in-Oil Microemulsion as an Appropriate Enzyme Carrier: The Model Case of Horseradish Peroxidase.

A novel biocompatible water-in-oil microemulsion was developed using nonionic surfactants and was investigated as a potential enzyme delivery system for pharmaceutical applications. The system was composed of isopropyl myristate/polysorbate 80 (Tween 80)/distilled monoglycerides/water/propylene glycol (PG), had a low total surfactant concentration (8.3% w/w), and was able to incorporate approximately 3% w/w aqueous phase containing horseradish peroxidase (HRP). Structural and activity aspects of the system were studied using a variety of techniques such as dynamic light scattering (DLS), electron paramagnetic resonance (EPR), and dynamic interfacial tension. The apparent hydrodynamic diameter of the empty droplets was calculated at about 37 nm. Different enzyme concentrations, ranging from 0.01 to 1.39 µM, were used for both DLS and EPR studies to effectively determine the localization of the macromolecule in the microemulsion. According to the results, for high enzyme concentrations, a participation of HRP in the surfactant monolayer of the microemulsion is evident. The number of reverse micelles in the microemulsion was defined by a theoretical model and was used to clarify how the enzyme concentration affects the number of empty and loaded reverse micelles. To assure that the system allows the enzyme to retain its catalytic activity, an oxidative reaction catalyzed by HRP was successfully carried out with the use of the model substrate 2,2'-azino-bis[3-ethylbenzothiazoline-6-sulfonic acid]. The influence of several parameters such as temperature, pH, and PG concentration was examined to optimize the reaction conditions, and a kinetic study was conducted revealing an ordered-Bi-Bi mechanism. Values of all kinetic parameters were determined. The release of the encapsulated enzyme was studied using an adequate receiver phase, revealing the effectiveness of the proposed microemulsion not only as a microreactor but also as a carrier for therapeutic biomolecules.

Oil from Mullet Roe Byproducts: Effect of Oil Extraction Method on Human Erythrocytes and Platelets.

Background: The valorization of byproducts to obtain high nutritional value foods is of utmost importance for our planet where the population is booming. Among these products are oils rich in ω-3 fatty acids produced from fishery byproducts. Recently, mullet roe oil from roe byproducts was produced that was rich in the ω-3 fatty acids eicosatetraenoic acid (EPA) and docosahexaenoic acid (DHA). Oils are customarily characterized for their composition and degree of oxidation but little is known of their biological effects, especially the effect of the extraction method. Methods: The purpose of this study was to evaluate the effects of freshly extracted mullet roe oil from mullet roe byproducts and the effect of the extraction method on human red blood cells (hRBCs) and platelets. To this end, the hemocompatibility (cytotoxicity), oxidative effects, and erythrocyte membrane changes were examined after 1 and 24 h of incubation. Antiplatelet effects were also assessed in vitro. Results: The expeller press oil extraction method and alcalase-assisted extraction produced the most biocompatible oils, as shown by hemocompatibility measurements and the absence of erythrocyte membrane alterations. Solvent extracts and protease-assisted extraction oils resulted in the rupture of red blood cells at different examined dilutions, creating hemolysis. Conclusions: It seems that the proper functioning of oil-erythrocyte interactions cannot be explained solely by ROS. Further investigations combining chemical analysis with oil-cell interactions could be used as an input to design high nutritional value oils using green extraction technologies. All samples exhibited promising antiplatelet and antiblood clotting effects in vitro.

Injection Molded PP Foams Using Food Ingredients for Food Packaging Applications.

A new approach to the creation of polypropylene (PP) based foaming materials was developed using food grade foaming agents that were coated on the PP pellets. More specifically, sodium bicarbonate and organic acids were used to coat PP pellets using either polyethyleneoxide (PEO) or lipid esters as coating stabilizers. In order to overcome the problem of the thermal decomposition of sodium bicarbonate at temperatures lower than the PP melting temperature, which makes the direct foaming during melt mixing impossible, the proposed methodology was proved quite efficient. Thus, new PP masterbatches were prepared, where the foaming agents were incorporated as coating at PP pellets at contents up to 10%, and initially used in Lab scale injection machines in order to find the best combination of materials that resulted in the production of foamed articles. Subsequently selected material combinations were tested in an industrial scale injection molding machine, where an optimization of the injection parameters was attempted. The outcome of this was the production of PP articles with significantly increased void fraction, up to 14%, decreased thermal conductivity, up to 20%, and various pore sizes as was observed via microscopic examination using SEM and CLSM.

Fractionation of a hydrocolloid emulsifier reclaimed from winery waste.

Complex hydrocolloids have been isolated and fractionated using a consecutive elution process, starting from winery waste. These extracts consist mainly of polysaccharidic populations and of smaller protein molecules and they exhibit emulsifying, thickening and texture-modifying activity. This work is a systematic study of these individual populations, as fractionated with preparative size exclusion chromatography (Prep-SEC) in terms of their chemical identity, surface properties, and emulsification behavior. The fractions have been characterized via SEC-MALLS, FTIR, DLS, zeta potential, and interfacial tension measurements. The results highlight the antagonistic and synergistic effects of the individual components of the above-mentioned complex natural material (winery waste extract) towards its emulsifying behavior, and provide a model for the kinetics of the evolution of a Pickering interfacial layer.

Insights on the Optical Properties of Poly(3,4-Ethylenedioxythiophene):Poly(styrenesulfonate) Formulations by Optical Metrology.

Poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) is among the most widely used polymers that are used as printed transparent electrodes for flexible Organic Electronic (OE) devices, such as Organic Photovoltaics (OPVs). The understanding of their optical properties and the correlation of the optical properties with their electronic properties and metallic-like behavior can lead to the optimization of their functionality as transparent electrodes in multilayer OE device architectures. In this work, we study the optical properties of different PEDOT:PSS formulations by non-destructive Spectroscopic Ellipsometry (SE), from the infrared to the far ultraviolet spectral regions. The optical response of PEDOT:PSS includes an intense optical absorption originated from the conductive part (PEDOT) at lower photon energies, whereas the electronic transition energies of the non-conductive PSS part have been measured at higher photon energies. Based on the different PEDOT:PSS formulations, the optical investigation revealed significant information on the relative contribution of conductive PEDOT and insulating PSS parts of the PEDOT:PSS formulation in the overall optical response, which can strongly impact the final device functionality and its optical transparency.

Effect of oleic acid on the properties of protein adsorbed layers at water/oil interfaces: An EPR study combined with dynamic interfacial tension measurements.

Multiphase food systems consist of complex interfacial layers where surface active molecules complete compete for adsorption and interact with one another affecting the interfacial properties and the behavior of the food systems involved. The present work focuses on the examination of interactions between proteins playing an which play an important role in milk-rich food systems (namely κ-casein and ß-lactoglobulin) and oleic acid (an anionic surfactant abundant in food systems) at the oil/water interface. An interesting feature of this system is that the molecules interacting at the interface originate from different phases and do not transfer/dissolve significantly to the other phase. The systems were examined using Electron Paramagnetic Resonance (EPR) spectroscopy and Dynamic Interfacial Tension measurements (dynamic interfacial tension and dynamic interfacial dilatational rheology). This combination of experimental methods provided the dynamics of adsorption at the interface and the mechanical properties of the interface allowing valuable insight on the interactions of the different molecules. The above information was coupled with direct information on fatty acid mobility in the oil bulk phase and indirect information on the degree of relative fatty acid/protein adsorption at the interface by EPR. Overall a synergistic effect of the protein and fatty acid on decreasing interfacial tension of the oil/water interface was evidenced. The fatty acid interacted differently with the random coil κ-casein and globular ß-lactoglobulin in terms of relative adsorption and in terms on its effect on mechanical properties. Thermal denaturation of ß-lactoglobulin affected the protein's interaction with the fatty acid due to conformational changes and exposure of non-polar sites.

Novel nanostructured biomaterials: implications for coronary stent thrombosis.

BACKGROUND: Nanomedicine has the potential to revolutionize medicine and help clinicians to treat cardiovascular disease through the improvement of stents. Advanced nanomaterials and tools for monitoring cell-material interactions will aid in inhibiting stent thrombosis. Although titanium boron nitride (TiBN), titanium diboride, and carbon nanotube (CNT) thin films are emerging materials in the biomaterial field, the effect of their surface properties on platelet adhesion is relatively unexplored. OBJECTIVE AND METHODS: In this study, novel nanomaterials made of amorphous carbon, CNTs, titanium diboride, and TiBN were grown by vacuum deposition techniques to assess their role as potential stent coatings. Platelet response towards the nanostructured surfaces of the samples was analyzed in line with their physicochemical properties. As the stent skeleton is formed mainly of stainless steel, this material was used as reference material. Platelet adhesion studies were carried out by atomic force microscopy and scanning electron microscopy observations. A cell viability study was performed to assess the cytocompatibility of all thin film groups for 24 hours with a standard immortalized cell line. RESULTS: The nanotopographic features of material surface, stoichiometry, and wetting properties were found to be significant factors in dictating platelet behavior and cell viability. The TiBN films with higher nitrogen contents were less thrombogenic compared with the biased carbon films and control. The carbon hybridization in carbon films and hydrophilicity, which were strongly dependent on the deposition process and its parameters, affected the thrombogenicity potential. The hydrophobic CNT materials with high nanoroughness exhibited less hemocompatibility in comparison with the other classes of materials. All the thin film groups exhibited good cytocompatibility, with the surface roughness and surface free energy influencing the viability of cells.