Confinement of Triple-Enzyme-Involved Antioxidant Cascade in Two-Dimensional Nanostructure.

Application of antioxidant enzymes in medical or industrial processes is limited due to their high sensitivity to environmental conditions. Incorporation of such enzymes in nanostructures provides a promising route to obtain highly efficient and robust biocatalytic system to scavenge reactive oxygen species (ROS). Here, this question was addressed by confinement of superoxide dismutase (SOD), horseradish peroxidase (HRP), and catalase (CAT) enzymes into nanostructures containing polyelectrolyte building blocks (alginate (Alg) and trimethyl chitosan (TMC)) and delaminated layered double hydroxide (dLDH) nanoparticle support. The nanocomposite possessed excellent structural and colloidal stability, while antioxidant tests revealed that the enzymes remained active upon immobilization and the developed composite greatly reduced intracellular oxidative stress in two-dimensional cell cultures. Moreover, it effectively prevented hydrogen peroxide-induced double stranded DNA breaks, which is a common consequence of oxidative stress. The results provide important tools to design complex nanostructures with multienzymatic antioxidant activities for ROS scavenging.

Application of Lacunarity for Quantification of Single Molecule Localization Microscopy Images.

The quantitative analysis of datasets achieved by single molecule localization microscopy is vital for studying the structure of subcellular organizations. Cluster analysis has emerged as a multi-faceted tool in the structural analysis of localization datasets. However, the results it produces greatly depend on the set parameters, and the process can be computationally intensive. Here we present a new approach for structural analysis using lacunarity. Unlike cluster analysis, lacunarity can be calculated quickly while providing definitive information about the structure of the localizations. Using simulated data, we demonstrate how lacunarity results can be interpreted. We use these interpretations to compare our lacunarity analysis with our previous cluster analysis-based results in the field of DNA repair, showing the new algorithm's efficiency.

Hierarchical Self-Assembly of Metal-Ion-Modulated Chitosan Tubules.

Soft materials such as gels or biological tissues can develop via self-assembly under chemo-mechanical forces. Here, we report the instantaneous formation of soft tubular structures with a two-level hierarchy by injecting a mixture of inorganic salt and chitosan (CS) solution from below into a reactor filled with alkaline solution. Folding and wrinkling instabilities occur on the originally smooth surface controlled by the salt composition and concentration. Liesegang-like precipitation patterns develop on the outer surface on a µm length scale in the presence of calcium chloride, while the precipitate particles are distributed evenly in the bulk as corroborated by X-ray µ-CT. On the other hand, barium hydroxide precipitates out only in the thin outer layer of the CS tubule when barium chloride is introduced into the CS solution. Independent of the concentration of the weakly interacting salt, an electric potential gradient across the CS membrane develops, which vanishes when the pH difference between the two sides of the membrane diminishes.

Influence of adsorption of ionic liquid constituents on the stability of layered double hydroxide colloids.

The influence of ionic liquid (IL) anions and cations on the charging and aggregation properties of layered double hydroxide (LDH) nanoparticles was systematically studied. Surface charge characteristics were explored using zeta potential measurements, while aggregation processes were followed in dynamic light scattering experiments in aqueous IL solutions. The results revealed that the aggregation rates of LDHs were sensitive to the composition of ILs leading to IL-dependent critical coagulation concentration (CCC) values being obtained. The origin of the interparticle forces was found to be electrostatic, in line with the classical Derjaguin-Landau-Verwey-Overbeek (DLVO) theory, as the experimental aggregation kinetics were in good agreement with the predicted data. The ion specific adsorption of IL anions led to different surface charge densities for LDHs, which decreased in the order Cl- > Br- > DCA- > SCN- > NO3- for counterions and BMIM+ > BMPYR+ > BMPY+ > BMPIP+ in the case of coions resulting in weaker electrical double layer repulsion in these sequences. Since van der Waals forces are always present and their strength does not depend significantly on the ionic strength, the CCC values decreased in the above order. The present results shed light on the importance of the interfacial arrangement of the IL constituent ions on the colloidal stability of particle dispersions and provide important information on the design of stable or unstable particle-ionic liquid systems.

Enhancing the yield of calcium carbonate precipitation by obstacles in laminar flow in a confined geometry.

Flow-driven precipitation experiments are performed in model porous media shaped within the confinement of a Hele-Shaw cell. Precipitation pattern formation and the yield of the reaction are investigated when borosilicate glass beads of different sizes are used in a mono-layer arrangement. The trend of the amount of precipitate produced in various porous media is estimated via visual observation. In addition, a new method is elaborated to complement such image analysis based results by titration experiments performed on gel-embedded precipitate patterns. The yield of confined porous systems is compared to experiments carried out in unsegmented reactors. It is found that the obstacles increase the amount of product and preserve its radial spatial distribution. The precipitate pattern is successfully conserved in a slightly cross-linked hydrogel matrix and its microstructure is examined using SEM. The spatial distribution of the precipitate across the cell gap is revealed using X-ray microtomography.

Catalytic antioxidant nanocomposites based on sequential adsorption of redox active metal complexes and polyelectrolytes on nanoclay particles.

An antioxidant nanocomposite was prepared by successive adsorption of redox active metal complexes (copper(ii)-bipyridyl and iron(iii)-citrate) and polyelectrolytes (poly(styrene sulfonate) and poly(diallyldimethyl ammonium)) on layered double hydroxide nanoclay. The experimental conditions were optimized in each preparation step and thus, the final composite formed highly stable colloids, i.e., excellent resistance against salt-induced aggregation was achieved. Due to the synergistic effect of the metal complexes, the developed composite showed remarkable activity in the dismutation of superoxide radicals, close to the one determined for the native superoxide dismutase enzyme. The obtained composite is highly selective for superoxide radical dismutation, while its activity in other antioxidant tests was close to negligible. Structural characterization of the composite revealed that the excellent superoxide radical scavenging ability originated from the advantageous coordination geometry around the copper(ii) center formed upon immobilization. The structure formed around the metal centers led to optimal redox features and consequently, to an improved superoxide dismutase-like activity. The catalytic antioxidant composite is a promising candidate to reduce oxidative stress in industrial manufacturing processes, where natural enzymes quickly lose their activity due to the harsh environmental conditions.

Out-of-plane auxetic nonwoven as a designer meta-biomaterial.

Biomaterials are porous and three-dimensional (3D) templates, which are used as biological substitutes in tissue engineering. Targeting the optimal design of biomaterials requires a synergy between mechanical, porous, mass transport, and biological properties. To address this challenge, we propose a non-periodic meta-biomaterial in the form of an out-of-plane auxetic nonwoven scaffold that possesses a 3D interconnected highly porous structure with remarkable mechanical properties corresponding to conventional nonwoven material. A design strategy of utilizing larger fiber diameters to enhance the porosity and permeability characteristics successfully devised the nonwoven scaffold with an extraordinary out-of-plane auxetic effect. In situ tensile-X-ray microcomputed tomography (microCT) analysis has been carried out to monitor the variation in the morphological characteristics.

Flexible planar supercapacitors by straightforward filtration and laser processing steps.

There is ever increasing demand for flexible energy storage devices due to the development of wearable electronics and other small electronic devices. The electrode flexibility is best provided by a special set of nanomaterials, but the required methodology typically consists of multiple steps and are designed just for the specific materials. Here, a facile and scalable method of making flexible and mechanically robust planar supercapacitors with interdigital electrode structure made of commercial carbon nanomaterials and silver nanowires is presented. The capacitor structure is achieved with vacuum filtration through a micropatterned contact mask and finished with simple laser processing steps. A maximum specific capacitance of 4 F cm-3 was measured with cyclic voltammetry at scan rate of 5 mV s-1. The reliability and charge transfer properties of devices were further investigated with galvanostatic charge-discharge measurements and electrochemical impedance spectroscopy, respectively. Furthermore, mechanical bending tests confirmed the devices have excellent mechanical integrity, and the deformations have no adverse effects on the electrochemical charge-discharge behavior and stability.

A Stimulus-Responsive Polymer Composite Surface with Magnetic Field-Governed Wetting and Photocatalytic Properties.

With the increasing demand for liquid manipulation and microfluidic techniques, surfaces with real-time tunable wetting properties are becoming the focus of materials science researches. In this study, we present a simple preparation method for a 0.5-4 µm carbonyl iron (carbonyl Fe) loaded polydimethylsiloxane (PDMS)-based magnetic composite coating with magnetic field-tailored wetting properties. Moreover, the embedded 6.3-16.7 wt.% Ag-TiO2 plasmonic photocatalyst (d~50 nm) content provides additional visible light photoreactivity to the external stimuli-responsive composite grass surfaces, while the efficiency of this photocatalytic behavior also turned out to be dependent on the external magnetic field. The inclusion of the photocatalyst introduced hierarchical surface roughness to the micro-grass, resulting in the broadening of the achievable contact and sliding angle ranges. The photocatalyst-infused coatings are also capable of catching and releasing water droplets, which alongside their multifunctional (photocatalytic activity and tunable wetting characteristics) nature makes surfaces of this kind the novel sophisticated tools of liquid manipulation.

Photocatalytic elimination of interfacial water pollutants by floatable photoreactive composite nanoparticles.

Disastrous oil spills cause severe environmental issues. The shortcomings of current cleaning methods for remediating oil have prompted the latest research drive to create intelligent nanoparticles that absorb oil. We, therefore, synthesized 197 ± 50 nm floatable photoreactive hybrid nanoparticles with Ag-TiO2 plasmonic photocatalyst (Eg = 3.08 eV) content to eliminate interfacial water pollutants, especially toluene-based artificial oil spill. We found that the composite particles have non-wetting properties in the aqueous media and float easily on the surface of the water due to the moderate hydrophobic nature (Θ = 113°) of the matrix of polystyrene, and these properties lead to elevated absorption of the interfacial organic pollutants (e.g., mineral oil). We showed that (28.5 mol%) divinylbenzene cross-linker content was required for adequate swelling capacity (2.15 g/g), whereas incorporated 15.8% Ag-TiO2 content in the swollen particles was enough for efficient photodegradation of the artificial oil spill under 150 min LED light (λmax = 405 nm) irradiation. The swollen polymer particles with embedded 32 ± 7 nm Ag-TiO2 content increase the efficiency of photooxidation by increased the direct contact between both the photocatalysts and the artificial oil spill. Finally, it was also presented that the composite particles destroy themselves: after approximately one and a half months of continuous LED light irradiation, the organic polymer component of the composite was almost completely (88.5%) photodegraded by the incorporated inorganic photocatalyst particles.

Layered Double Hydroxide Nanoparticles to Overcome the Hydrophobicity of Ellagic Acid: An Antioxidant Hybrid Material.

Ellagic acid (EA), a polyphenolic antioxidant of poor water solubility, was intercalated into biocompatible layered double hydroxide (LDH) nanoparticles by the coprecipitation method. Structural investigation of the composite revealed that the lactone bonds split under the synthetic experimental conditions, and EA was transformed to 4,4',5,5',6,6'-hexahydroxydiphenic acid during intercalation. To improve the surface properties of the EA-LDH composite, the samples were treated with different organic solvents. The antioxidant activity of the LDH hybrids was assessed in test reactions. Most of the obtained hybrids showed antioxidant activity comparable to the one of the free EA indicating that the spontaneous structural transformation upon immobilization did not change the efficiency in radical scavenging. Treatments with organic solvents influenced the activities of the materials remarkably. The main advantage of the immobilization procedure is that the products can be applied in aqueous samples in high concentrations overcoming the problem related to the low solubility of EA in water. The developed composites of high antioxidant content can be applied as efficient reactive oxygen species scavenging materials during biomedical treatments or industrial manufacturing processes.

Fast optical method for characterizing plasmonic nanoparticle adhesion on functionalized surfaces.

In this paper, a rapid optical method for characterizing plasmonic (gold) nanoparticle (AuNP) adhesion is presented. Two different methods were used for AuNP preparation: the well-known Turkevich method resulted in particles with negative surface charge; for preparing AuNPs with positive surface charge, stainless steel was used as reducing agent. The solid surface for adhesion was provided by a column packed with pristine or surface-modified glass beads. The size of the nanoparticles was studied by transmission electron microscopy (TEM) and small-angle X-ray scattering (SAXS); the surface charge of the components was determined by streaming potential measurements. The characterization of adhesion was performed in a flow system by UV-Vis spectroscopy. During the adhesion experiments, the role of the surface charge, the particle size, and the pH were studied, as well as the adhered amount of gold nanoparticles and the surface coverage values. The latter was estimated by theoretical calculations and defined by the quotient of the measured and the maximal adhered amount of nanoparticles, which could be determined by the cross-sectional area of the NPs and the specific surface area of the glass beads. The results are verified by the polarization reflectometric interference spectroscopy (PRIfS) method: silica nanoparticles with diameters of a few hundred (d~450) nanometers were immobilized on the surface of glass substrate by the Langmuir-Blodgett method, the surface was modified similar to the 3D (continuous flow packed column) system, and gold nanoparticles from different pH solutions were adhered during the measurements. These kinds of modified surfaces allow the investigation of biomolecule adsorption in the same reflectometric setup. Graphical abstract.

Magnetic-Field-Manipulated Growth of Flow-Driven Precipitate Membrane Tubes.

Chemobrionics is an emerging scientific field focusing on the coupling of chemical reactions and different forms of motion, that is, transport processes. Numerous phenomena appearing in various gradient fields, for example, pH, concentration, temperature, and so on, are thoroughly investigated to mimic living systems in which spatial separation plays a major role in proper functioning. In this context, chemical garden experiments have received increased attention because they inherently involve membrane formation and various transport processes. In this work, a noninvasive external magnetic field was applied to gain control over the directionality of membrane structures obtained by injecting one reactant solution into the other in a three-dimensional domain. The geometry of the resulted patterns was quantitatively characterized as a function of the injection rate and the magnitude of magnetic induction. The magnetic field was proven to influence the microstructure of precipitate tubes by diminishing spatial defects.

Preparation of novel tissue acidosis-responsive chitosan drug nanoparticles: Characterization and in vitro release properties of Ca2+ channel blocker nimodipine drug molecules.

The pH-responsive intelligent drug release facility of hydrophobically modified chitosan nanoparticles (Chit NPs) (d = 5.2 ±â€¯1.1 nm) was presented in the case of poorly water soluble Ca2+ channel blocker nimodipine (NIMO) drug molecules. The adequate pH-sensitivity, i.e. the suitable drug carrier properties of the initial hydrophilic Chit were achieved by reductive amination of Chit with hexanal (C6-) and dodecanal (C12-) aldehydes. The successful modifications of the macromolecule were evidenced via FTIR measurements: the band appearing at 1412 cm-1 (CN stretching in aliphatic amines) in the cases of the hydrophobically modified Chit samples shows that the CN bond successfully formed between the Chit and the aldehydes. Hydrophobization of the polymer unambiguously led to lower water contents with lower intermolecular interactions in the prepared hydrogel matrix: the initial hydrophilic Chit has the highest water content (78.6 wt%) and the increasing hydrophobicity of the polymer resulted in decreasing water content (C6-chit.: 74.2 wt% and C12-chit.: 47.1 wt%). Furthermore, it was established that the length of the side chain of the aldehyde influences the pH-dependent solubility properties of the Chit. Transparent homogenous polymer solution was obtained at lower pH, while at higher pH the formation of polymer (nano)particles was determined and the corresponding cut-off pH values showed decreasing tendency with increasing hydrophobic feature (pH = 7.47, 6.73 and 2.49 for initial Chit, C6-chit and C12-chit, respectively). Next the poorly water soluble NIMO drug was encapsulated with the C6-chit with adequate pH-sensitive properties. The polymer-stabilized NIMO particles with 10 wt% NIMO content resulted in stable dispersion in aqueous phase, the formation of polymer shell increased in the water solubility/dispersibility of the initial hydrophobic drug. According to the drug release experiments, we clearly confirmed that the encapsulated low crystallinity NIMO drug remained closed in the polymer NPs at normal tissue pH (pH = 7.4, PBS buffer, physiological condition) but at pH < 6.5 which is typical for seriously ischemic brain tissue, 93.6% of the available 0.14 mg/ml NIMO was released into the buffer solution under 8 h release time. According to this in vitro study, the presented pH-sensitive drug carrier system could be useful to selectively target ischemic brain regions characterized by acidosis, to achieve neuroprotection at tissue zones at risk of injury, without any undesirable side effects caused by systemic drug administration.

A redox strategy to tailor the release properties of Fe(III)-alginate aerogels for oral drug delivery.

Iron(III)-crosslinked alginate aerogel beads (d = 3-5 mm) were prepared and loaded with ibuprofen by using the technique of adsorptive deposition from supercritical CO2. Additional formulations were prepared where the aerogels were co-impregnated by ibuprofen and ascorbic acid. The release of ibuprofen from the Fe(III)-alginate is much faster in pH = 7.4 (PBS) than in pH = 2.0 (HCl), which can be explained by the faster dissolution and higher swelling of the alginate matrix in PBS. By decreasing the size of the beads and using a higher G content alginate the release rate could be slightly increased. A marked acceleration of drug release was achieved in both HCl and PBS by incorporating ascorbic acid into the Fe(III)-alginate aerogel preparations. The explanation is that in aqueous media ascorbic acid in situ reduces the crosslinking Fe(III) to Fe(II). The latter does not interact strongly with alginate, which promotes the hydration of the chains, thus the erosion and dissolution of the carrier matrix.

Spherical LDH-Ag°-montmorillonite heterocoagulated system with a pH-dependent sol-gel structure for controlled accessibility of AgNPs immobilized on the clay lamellae.

Aqueous suspensions of spherical ZnMgAl-layered double hydroxides [LDH(sph)] and antibacterial silver nanoparticles (AgNPs) deposited on the lamellae of montmorillonite were used for the synthesis of composites, which behave like coherent gels at low pH (≲4.5) and incoherent sols at higher pH (≳4.5). The composition of the composite was chosen as LDH(sph)/Ag°-montm. = 25:75 wt % in order to ensure a sol-gel transition that can also be characterized by viscometry. This pH-sensitive heterocoagulated system consisting of oppositely charged colloid particles was suitable for the release of antimicrobial AgNPs immobilized on the clay lamellae via a pH-controlled gel-sol transition. The heterocoagulation process was also characterized by surface charge titration measurements. Spherical LDH/Ag°-montmorillonite composite samples were identified by X-ray diffraction (XRD) measurements. The morphological properties of the composites were studied, and the presence of the heterocoagulated structure was confirmed by scanning electron microscopy (SEM). The nanoscale structure of the LDH(sph)-Ag°-montmorillonite composite obtained was also verified by small-angle X-ray scattering (SAXS), and the rheological characteristics were studied at various pH values. The viscosity and yield value of the composite decreased by an order of magnitude upon increasing the pH from 3.0 to 5.5. The sol-gel transition of the composite suspension was reversible in the previously mentioned pH range.

Optical and structural properties of protein/gold hybrid bio-nanofilms prepared by layer-by-layer method.

Lysozyme/gold thin layers were prepared by layer-by-layer (LbL) self-assembly method. The build-up of the films was followed by UV-vis-absorbance spectra, quartz crystal microbalance (QCM) and surface plasmon resonance (SPR) techniques. The structural property of films was examined by X-ray diffraction (XRD) measurements, while their morphology was studied by scanning electron microscopy (SEM) and atomic force microscopy (AFM). It was found that gold nanoparticles (NPs) had cubic crystalline structure, the primary particles form aggregates in the thin layer due to the presence of lysozyme molecules. The UV-vis measurements prove change in particle size while the colour of the film changes from wine-red to blue. The layer thickness of films was determined using the above methods and the loose, porous structure of the films explains the difference in the results. The vapour adsorption property of hybrid layers was also studied by QCM using different saturated vapours and ammonia gas. The lysozyme/Au films were most sensitive for ammonia gas among the tested gases/vapours due to the strongest interaction between the functional groups of the protein.

Structural, optical, and adsorption properties of ZnO(2)/poly(acrylic acid) hybrid thin porous films prepared by ionic strength controlled layer-by-layer method.

ZnO(2)/poly(acrylic acid) sandwich structures were prepared by layer-by-layer (LbL) self-assembly. The structure and optical behavior of the hybrid films were controlled by changing the surface charge and conformation of the poly(acrylic acid). The buildup of the films was followed by UV-vis absorption and reflection spectroscopy, atomic force microscopy (AFM), X-ray diffraction (XRD), and quartz crystal microbalance (QCM) measurements. It was found that the ionic strength of the polymer solution had a great influence on the film thickness which, in turn, affected the optical properties. The water vapor adsorption isotherms of the films determined by QCM showed an adsorption hysteresis characteristic of porous thin layer structures. The adsorption of water molecules inside the films changed the effective refractive index resulting in a change of the reflection properties. This phenomenon is shown to be exploited for the application of the films as optical sensors. The polarizability of water molecules in the adsorption layer was also determined. It was found that polarization of water molecules in the adsorption layer is much lower than in the liquid water when the surface coverage (Theta) is low.