Improved virus removal in ceramic depth filters modified with MgO.

Ceramic filters, working on the depth filtration principle, are known to improve drinking water quality by removing human pathogenic microorganisms from contaminated water. However, these microfilters show no sufficient barrier for viruses having diameters down to 20 nm. Recently, it was shown that the addition of positively charged materials, for example, iron oxyhydroxide, can improve virus removal by adsorption mechanisms. In this work, we modified a common ceramic filter based on diatomaceous earth by introducing a novel virus adsorbent material, magnesium oxyhydroxide, into the filter matrix. Such filters showed an improved removal of about 4-log in regard to bacteriophages MS2 and PhiX174. This is explained with the electrostatic enhanced adsorption approach that is the favorable adsorption of negatively charged viruses onto positively charged patches in an otherwise negatively charged filter matrix. Furthermore, we provide theoretical evidence applying calculations according to Derjaguin-Landau-Verwey-Overbeek theory to strengthen our experimental results. However, modified filters showed a significant variance in virus removal efficiency over the course of long-term filtration experiments with virus removal increasing with filter operation time (or filter aging). This is explained by transformational changes of MgO in the filter upon contact with water. It also demonstrates that filter history is of great concern when filters working on the adsorption principles are evaluated in regard to their retention performance as their surface characteristics may alter with use.

Virus removal in ceramic depth filters based on diatomaceous earth.

Ceramic filter candles, based on the natural material diatomaceous earth, are widely used to purify water at the point-of-use. Although such depth filters are known to improve drinking water quality by removing human pathogenic protozoa and bacteria, their removal regarding viruses has rarely been investigated. These filters have relatively large pore diameters compared to the physical dimension of viruses. However, viruses may be retained by adsorption mechanisms due to intermolecular and surface forces. Here, we use three types of bacteriophages to investigate their removal during filtration and batch experiments conducted at different pH values and ionic strengths. Theoretical models based on DLVO-theory are applied in order to verify experimental results and assess surface forces involved in the adsorptive process. This was done by calculation of interaction energies between the filter surface and the viruses. For two small spherically shaped viruses (MS2 and PhiX174), these filters showed no significant removal. In the case of phage PhiX174, where attractive interactions were expected, due to electrostatic attraction of oppositely charged surfaces, only little adsorption was reported in the presence of divalent ions. Thus, we postulate the existence of an additional repulsive force between PhiX174 and the filter surface. It is hypothesized that such an additional energy barrier originates from either the phage's specific knobs that protrude from the viral capsid, enabling steric interactions, or hydration forces between the two hydrophilic interfaces of virus and filter. However, a larger-sized, tailed bacteriophage of the family Siphoviridae was removed by log 2 to 3, which is explained by postulating hydrophobic interactions.

Modification of ceramic microfilters with colloidal zirconia to promote the adsorption of viruses from water.

The purpose of this study was to test the feasibility of modifying commercial microporous ceramic bacteria filters to promote adsorption of viruses. The internal surface of the filter medium was coated with ZrO(2) nanopowder via dip-coating and heat-treatment in order to impart a filter surface charge opposite to that of the target viruses. Streaming potential measurements revealed a shift in the isoelectric point from pH <3 to between pH 5.5 and 9, respectively. While the base filter elements generally exhibited only 75% retention with respect to MS2 bacteriophages, the modified elements achieved a 7log removal (99.99999%) of these virus-like particles. The coating process also increased the specific surface area of the filters from approximately 2m(2)/g to between 12.5 and 25.5m(2)/g, thereby also potentially increasing their adsorption capacity. The results demonstrate that, given more development effort, the chosen manufacturing process has the potential to yield effective virus filters with throughputs superior to those of current virus filtration techniques.

Investigating the time-dependent zeta potential of wood surfaces.

This work reports on streaming potential measurements through natural capillaries in wood and investigates the cause of a time-dependent zeta potential measured during the equilibration of wood cell-walls with an electrolyte solution. For the biomaterial, this equilibration phase takes several hours, which is much longer than for many other materials that have been characterized by electrokinetic measurements. During this equilibration phase the zeta potential magnitude is decaying due to two parallel mechanisms: (i) the swelling of the cell-wall which causes a dimensional change reducing the charge density at the capillary interface; (ii) the transport of ions from the electrolyte solution into the permeable cell-wall which alters the electrical potential at the interface by internal charge compensation. The obtained results demonstrate the importance of equilibration kinetics for an accurate determination of the zeta potential, especially for materials that interact strongly with the measurement electrolyte. Moreover, the change in zeta potential with time can be correlated with the bulk swelling of wood if the effect of electrolyte ion diffusion is excluded. This study shows the potential of streaming potential measurements of wood, and possibly of other hygroscopic and nanoporous materials, to reveal kinetic information about their interaction with liquids, such as swelling and ion uptake.

Application of response surface methodology to tailor the surface chemistry of electrospun chitosan-poly(ethylene oxide) fibers.

Chitosan is a promising biocompatible polymer for regenerative engineering applications, but its processing remains challenging due to limited solubility and rigid crystalline structure. This work represents the development of electrospun chitosan/poly(ethylene oxide) blend nanofibrous membranes by means of a numerical analysis in order to identify and tailor the main influencing parameters with respect to accessible surface nitrogen functionalities which are of importance for the biological activity as well as for further functionalization. Depending on the solution composition, both gradient fibers and homogenous blended fiber structures could be obtained with surface nitrogen concentrations varying between 0 and 6.4%. Response surface methodology (RSM) revealed chitosan/poly(ethylene oxide) ratio and chitosan molecular weight as the main influencing factors with respect to accessible nitrogen surface atoms and respective concentrations. The model showed good adequacy hence providing a tool to tailor the surface properties of chitosan/poly(ethylene oxide) blends by addressing the amount of accessible chitosan.

Humic acid desorption from a positively charged nanocellulose surface.

Trimethyl-ammonium functionalized cellulose nanofibers (CNF) display a positively charged surface over a wide pH and ionic strength range, and constitute therefore not only an interesting biobased sorbent material, but also a positively charged model surface to investigate sorption phenomena. In the current work, CNF were totally or partially covered by humic acid (HA) at pH ∼6, and the desorption process of HA from CNF was thereafter studied in batch and continuous filtration experiments at various pH and ionic strength. It is found that the desorbed amount of HA increases and the kinetic of desorption is faster via increasing the pH and ionic strength of the solution. Moreover, the desorption is affected by the extent of CNF coverage with HA and by the presence of free HA in solution. The present work gives insights into the interaction of HA with permanently positively charged surfaces and was successfully exploited for the removal of HA from CNF filters for regeneration allowing multiple filter utilization.

High-Resolution Adhesion Mapping of the Odd-Even Effect on a Layer-by-Layer Coated Biomaterial by Atomic-Force-Microscopy.

The adhesion behavior of polyelectrolyte multilayers consisting of poly(diallyldimethylammonium chloride), PDDA, and poly(styrenesulfonate), PSS, toward a silicon AFM tip was studied during their build-up on wood, a chemically heterogeneous, micrometer rough biomaterial and compared with a nanometer rough substrate, namely quartz. The atomic force microscopy-based force mapping approach generated high-resolution topography-, and adhesion maps within the first bilayers, which point toward a homogeneous layer-by-layer build-up on the biomaterial surface, and therefore indicate an even charge distribution. By analyzing the force-distance curves in every pixel of the mapping, new insights into the specific interactions of the polyelectrolyte multilayers at the surface were achieved. The characteristic odd-even effect of polyelectrolyte multilayers cannot only be determined on quartz, but also on the biomaterial wood, however, only after an offset of two bilayers. This is potentially due to the specific roughness and charge of wood in comparison to commonly used quartz.

Layer-by-layer deposition on a heterogeneous surface: Effect of sorption kinetics on the growth of polyelectrolyte multilayers.

Surface functionalization by means of controlled deposition of charged polymers or nanoparticles using the layer-by-layer (LbL) approach has been used to modify mostly engineered materials with well-defined surface chemistry and morphology. In this regard, natural and inhomogeneous interfaces have gained very little attention. Furthermore, natural substrates are susceptible to alterations by factors commonly used to control the growth of multilayers, such as pH, temperature and ionic strength. Here, we study the impact of sorption kinetics of a bilayer system (Poly(diallyldimethylammonium chloride) (PDDA) and Poly(sodium 4-styrenesulfonate) (PSS)) on a natural heterogeneous wood surface at neutral pH, without salt addition, on the multilayer buildup. To overcome analytical limitations we introduce a complementary approach based on UV reflectance spectroscopy, atomic force microscopy (AFM) and zeta potential measurements. Compared to immersion times used for ideal substrates, we found that a high surface coverage requires relatively long immersion, approximately 30min, into polyelectrolyte solutions, while a sufficient removal of polyelectrolyte excess during the washing step, requires even longer, about 100min. Based on these findings, we show that film growth can be controlled kinetically. Long immersion times provide well-defined and regular multilayers. The obtained data points to specific requirements to be considered when LbL treatments are applied to rough, porous and heterogeneous surfaces, and thereby sets a basis for a successful transfer of various surface functionalization approaches already shown on ideal surfaces.

Avoiding drying-artifacts in transmission electron microscopy: Characterizing the size and colloidal state of nanoparticles.

Standard transmission electron microscopy nanoparticle sample preparation generally requires the complete removal of the suspending liquid. Drying often introduces artifacts, which can obscure the state of the dispersion prior to drying and preclude automated image analysis typically used to obtain number-weighted particle size distribution. Here we present a straightforward protocol for prevention of the onset of drying artifacts, thereby allowing the preservation of in-situ colloidal features of nanoparticles during TEM sample preparation. This is achieved by adding a suitable macromolecular agent to the suspension. Both research- and economically-relevant particles with high polydispersity and/or shape anisotropy are easily characterized following our approach (http://bsa.bionanomaterials.ch), which allows for rapid and quantitative classification in terms of dimensionality and size: features that are major targets of European Union recommendations and legislation.

Characterization of Pores in Dense Nanopapers and Nanofibrillated Cellulose Membranes: A Critical Assessment of Established Methods.

Nanofibrillated cellulose (NFC) is a natural fibrous material that can be readily processed into membranes. NFC membranes for fluid separation work in aqueous medium, thus in their swollen state. The present study is devoted to a critical investigation of porosity, pore volume, specific surface area, and pore size distribution of dry and wet NFC nanopapers, also known as membranes, with various established techniques, such as electron microscopy, helium pycnometry, mercury intrusion, gas adsorption (N2 and Kr), and thermoporometry. Although these techniques can be successfully applied to inorganic materials (e.g., mesoporous silica), it is necessary to appraise them for organic and hydrophilic products such as NFC membranes. This is due to different phenomena occurring at the materials interfaces with the probing fluids. Mercury intrusion and gas adsorption are often used for the characterization of porosity-related properties; nevertheless, both techniques characterize materials in the dry state. In parallel, thermoporometry was employed to monitor the structure changes upon swelling, and a water permeance test was run to show the accessibility of the membranes to fluids. For the first time, the methods were systematically screened, and we highlighted the need of uniform sample treatments prior to the measurements (i.e., sample cutting and outgassing protocols) in order to harmonize results from the literature. The need for revising the applicability range of mercury intrusion and the inappropriateness of nitrogen adsorption were pointed out. We finally present a table for selecting the most appropriate method to determine a desired property and propose guidelines for results interpretation from which future users could profit.

Different endocytotic uptake mechanisms for nanoparticles in epithelial cells and macrophages.

Precise knowledge regarding cellular uptake of nanoparticles is of great importance for future biomedical applications. Four different endocytotic uptake mechanisms, that is, phagocytosis, macropinocytosis, clathrin- and caveolin-mediated endocytosis, were investigated using a mouse macrophage (J774A.1) and a human alveolar epithelial type II cell line (A549). In order to deduce the involved pathway in nanoparticle uptake, selected inhibitors specific for one of the endocytotic pathways were optimized regarding concentration and incubation time in combination with fluorescently tagged marker proteins. Qualitative immunolocalization showed that J774A.1 cells highly expressed the lipid raft-related protein flotillin-1 and clathrin heavy chain, however, no caveolin-1. A549 cells expressed clathrin heavy chain and caveolin-1, but no flotillin-1 uptake-related proteins. Our data revealed an impeded uptake of 40 nm polystyrene nanoparticles by J774A.1 macrophages when actin polymerization and clathrin-coated pit formation was blocked. From this result, it is suggested that macropinocytosis and phagocytosis, as well as clathrin-mediated endocytosis, play a crucial role. The uptake of 40 nm nanoparticles in alveolar epithelial A549 cells was inhibited after depletion of cholesterol in the plasma membrane (preventing caveolin-mediated endocytosis) and inhibition of clathrin-coated vesicles (preventing clathrin-mediated endocytosis). Our data showed that a combination of several distinguishable endocytotic uptake mechanisms are involved in the uptake of 40 nm polystyrene nanoparticles in both the macrophage and epithelial cell line.