Effect of divalent versus monovalent cations on the MS2 retention capacity of amino-functionalized ceramic filters.

Ceramic capillary membranes conditioned for virus filtration via functionalization with n-(3-trimethoxysilylpropyl)diethylenetriamine (TPDA) are analyzed with respect to their virus retention capacity when using feed solutions based on monovalent and divalent salts (NaCl, MgCl2). The log reduction value (LRV) by operating in dead-end mode using the model bacteriophage MS2 with a diameter of 25 nm and an IEP of 3.9 is as high as 9.6 when using feeds containing MgCl2. In contrast, a lesser LRV of 6.4 is observed for feed solutions based on NaCl. The TPDA functionalized surface is simulated at the atomistic scale using explicit-solvent molecular dynamics in the presence of either Na+ or Mg2+ ions. Computational prediction of the binding free energy reveals that the Mg2+ ions remain preferentially adsorbed at the surface, whereas Na+ ions form a weakly bound dissolved ionic layer. The charge shielding between surface and amino groups by the adsorbed Mg2+ ions leads to an upright orientation of the TPDA molecules as opposed to a more tilted orientation in the presence of Na+ ions. The resulting better accessibility of the TPDA molecules is very likely responsible for the enhanced virus retention capacity using a feed solution with Mg2+ ions.

An evaluation of colloidal and crystalline properties of CaCO3 nanoparticles for biological applications.

Biodegradable calcium carbonate carriers are a promising and safe nanoparticle platform which might enable various applications as an engineered nanomaterial in health care, food and cosmetics. However, engineered nanoparticles can exhibit new forms of toxicity that must be carefully evaluated before being widely adopted in consumer products or novel drug delivery systems. To this end, we studied four common calcium carbonate particle systems (calcite nanoparticles, amorphous sub-micrometer and vaterite sub-micrometer and micrometer particles) and compared their behavior in biological medium and in cell culture experiments. The thermodynamically stable calcite phase is shown to maintain its morphological features as no phase transformation occurs. Size- and time-dependent phase transformation of the less stable vaterite particles are observed within 96h in cell medium. The protein serum albumin can be an effective inhibitor of phase-transition and it is shown to improve colloidal stability. The impact of the biological environment goes beyond protein-corona formation, as we observed rapid dissolution of amorphous particles in high ionic strength cell medium, but not in Millipore water. Cellular responses of human osteoblasts against CaCO3 particles indicate that increased intracellular calcium ions improve viability and that particle internalization is not size-dependent. Useful insights for designing CaCO3-based delivery systems are provided and also corroborate to the idea that intrinsic material properties as well as environmental conditions are of relevance for the successful implementation of dispersed CaCO3 particles in drug delivery systems and in other applications.

Chitosan supraparticles with fluorescent silica nanoparticle shells and nanodiamond-loaded cores.

Spherical colloidal structures with a sparsely or densely packed shell of nanoparticles have been the subject of intense research efforts for more than a decade. Research has focused on the utilization of aqueous, soft, or solid cores mainly on the micron-scale. A synthesis route that combines a stabilizing biopolymer core in combination with small (15 nm) or ultrasmall (5 nm) nanoparticles templated on the surface is still missing and pivotal to push these structures further into bionanotechnological applications. Here, we present core-shell supraparticles with a nanodiamond-loaded chitosan core and a shell assembled from fluorescent silica nanoparticles with controlled packing density which feature final sizes significantly below one micrometer. The synthesis route is highly versatile and represents a general approach for the synthesis of sophisticated supraparticles with shells formed from distinct nanoparticle shells and biopolymer cores loaded with selective nanoparticles.

Biodegradable and bioactive porous polymer/inorganic composite scaffolds for bone tissue engineering.

Biodegradable polymers and bioactive ceramics are being combined in a variety of composite materials for tissue engineering scaffolds. Materials and fabrication routes for three-dimensional (3D) scaffolds with interconnected high porosities suitable for bone tissue engineering are reviewed. Different polymer and ceramic compositions applied and their impact on biodegradability and bioactivity of the scaffolds are discussed, including in vitro and in vivo assessments. The mechanical properties of today's available porous scaffolds are analyzed in detail, revealing insufficient elastic stiffness and compressive strength compared to human bone. Further challenges in scaffold fabrication for tissue engineering such as biomolecules incorporation, surface functionalization and 3D scaffold characterization are discussed, giving possible solution strategies. Stem cell incorporation into scaffolds as a future trend is addressed shortly, highlighting the immense potential for creating next-generation synthetic/living composite biomaterials that feature high adaptiveness to the biological environment.

Change of zeta potential of biocompatible colloidal oxide particles upon adsorption of bovine serum albumin and lysozyme.

The amounts of negatively charged bovine serum albumin and positively charged lysozyme adsorbed on alumina, silica, titania, and zirconia particles (diameters 73 to 271 nm) in aqueous suspensions are measured. The adsorbed proteins change the zeta potentials and the isoelectric points (IEP) of the oxide particles. The added to adsorbed protein ratios at pH 7.5 are compared with the protein treated particle zeta potentials. It is found that the amounts of adsorbed proteins on the alumina, silica, and titania (but not on the zirconia) particle surfaces are highly correlated with the zeta potential. For the slightly less hydrophilic zirconia particles high amounts of protein adsorption are observed even under repulsive electrostatic conditions. One reason could be that the hydrophobic effect plays a more important role for zirconia than electrostatic interaction.

Coacervate-directed synthesis of CaCO3 microcarriers for pH-responsive delivery of biomolecules.

We report the synthesis of pH-responsive microcarriers via the combination of complex coacervation and mineralization of calcium carbonate (CaCO3). Positively and negatively charged proteins (bovine serum albumin (BSA) and lysozyme (LSZ)) form electrostatic complexes with poly(acrylic acid) sodium salt (PAANa) and calcium ions in an aqueous solution, leading to the formation of spherical coacervate droplets. By the addition of sodium carbonate, the protein-loaded droplets are mineralized into stable CaCO3 microcarriers. Since this inorganic material exhibits high solubility in acids, the release of protein from the carriers can be controlled via the pH of the environment. The process results in the successful generation of bulk amounts of monodisperse and colloidally stable microspheres with diameters as small as 300 nm. As the entire synthesis takes place under aqueous conditions, coacervate-directed encapsulation is suitable for sensitive active agents. Accordingly, the method presents a promising approach to synthesize pH-responsive microcarriers for drug delivery applications.

Orientation of human osteoblasts on hydroxyapatite-based microchannels.

The effect of calcium phosphate-based microchannels on the growth and orientation of human osteoblast cells is investigated in this study. As substrates, hydroxyapatite-based microchannels with high contouring accuracy were fabricated by a novel micro-moulding technique. Microchannels obtained by this method featured widths ranging from 16.0±0.7 to 76.6±1.4 µm and depths from 7.9±0.8 to 15.5±1.3 µm. Surface and contour characterization was carried out using X-ray diffraction analysis, scanning electron microscopy imaging and 3D-confocal profilometry. Cell activity and alignment on microchannels with different widths were determined after 1 and 3 days by photometric spectroscopy and fluorescence microscopic imaging and statistically analysed by Tukey's multiple comparison test. On days 1 and 3 for microchannels of width 16 and 30 µm, 70-80% of the osteoblasts oriented within an angular range of 0-15° relative to the microchannel direction. Interestingly, only 20% of the cells grew inside the microchannels for channel widths of 16 and 30 µm. Substrates with channel widths of 45, 65 and 76 µm allowed ∼40% of the cells to grow inside. The depth of the microchannel showed hardly any significant impact. All micropatterned surfaces provoked a good cell attachment, as flat and spread cell morphologies with lamellipodiae and filopodiae could already be observed after 1 day. The effect of the microchannels on osteoblast activity was determined using the colorimetric WST-1 assay. In addition, the cell differentiation was assessed by collagen type I staining. The cell activity obtained by WST-1 assay differed insignificantly for all micropatterned samples of various widths and depths. The assessment of collagen type I yielded the same amounts for all micropatterned samples after 1, 3 and 7 days. In summary, the microchannel width of HA-based patterns has a distinct effect on the directed growth of human osteoblast cells, allowing novel design strategies for surfaces such as dental implants.

Collagen release kinetics of surface functionalized 45S5 Bioglass-based porous scaffolds.

A highly interconnected porous scaffold made from 45S5 Bioglass was fabricated by the polymer replica technique and surface functionalized for protein immobilization. Subsequently rat-tail collagen type I was immobilized on the scaffolds. The protein and ion release rates were determined by UV-vis spectroscopy and ion chromatography, respectively, and the impact on hydroxyapatite (HA) formation on the scaffolds upon immersion in SBF was evaluated. It was discovered that the surface functionalization enhanced the stability of the collagen attachment and stability against the increment of pH in a biological environment, resulting in similar collagen release kinetics in solutions of different pH values. Without the surface modification, collagen release was considerably expedited by the increment of pH in a surrounding solution. It was also found that the collagen immobilization does not effect the formation of carbonated HA on the scaffold surface. The stable collagen attachment to the functionalized scaffold makes this approach potentially suitable for improving cell attachment and thus for enhancing the application potential of the scaffold in tissue engineering.

The surface functionalization of 45S5 Bioglass-based glass-ceramic scaffolds and its impact on bioactivity.

The first and foremost function of a tissue engineering scaffold is its role as a substrate for cell attachment, and their subsequent growth and proliferation. However, cells do not attach directly to the culture substrate; rather they bind to proteins that are adsorbed to the scaffold's surface. Like standard tissue culture plates, tissue engineering scaffolds can be chemically treated to couple proteins without losing the conformational functionality; a process called surface functionalization. In this work, novel highly porous 45S5 Bioglass-based scaffolds have been functionalized applying 3-AminoPropyl-TriethoxySilane (APTS) and glutaraldehyde (GA) without the use of organic solvents. The efficiency and stability of the surface modification was assessed by X-ray photoemission spectroscopy (XPS). The bioactivity of the functionalized scaffolds was investigated using simulated body fluid (SBF) and characterized by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS) and X-ray diffraction (XRD). It was found that the aqueous heat-treatment applied at 80 degrees C for 4 hrs during the surface functionalization procedure accelerated the structural transition of the crystalline Na2Ca2Si3O9 phase, present in the original scaffold structure as a result of the sintering process used for fabrication, to an amorphous phase during SBF immersion. The surface functionalized scaffolds exhibited an accelerated crystalline hydroxyapatite layer formation upon immersion in SBF caused by ion leaching and the increased surface roughness induced during the heat treatment step. The possible mechanisms behind this phenomenon are discussed.