Self-Organization of Motor-Propelled Cytoskeletal Filaments at Topographically Defined Borders

Self-organization phenomena are of critical importance in living organisms and of great interest to exploit in nanotechnology. Here we describe in vitro self-organization of molecular motor-propelled actin filaments, manifested as a tendency of the filaments to accumulate in high density close to topographically defined edges on nano- and microstructured surfaces. We hypothesized that this “edge-tracing” effect either (1) results from increased motor density along the guiding edges or (2) is a direct consequence of the asymmetric constraints on stochastic changes in filament sliding direction imposed by the edges. The latter hypothesis is well captured by a model explicitly defining the constraints of motility on structured surfaces in combination with Monte-Carlo simulations [cf. Nitta et al. (2006)] of filament sliding. In support of hypothesis 2 we found that the model reproduced the edge tracing effect without the need to assume increased motor density at the edges. We then used model simulations to elucidate mechanistic details. The results are discussed in relation to nanotechnological applications and future experiments to test model predictions

Nanometer table-top proximity x-ray lithography with liquid-target laser-plasma source

A compact laser-plasma proximity x-ray lithography system suitable for laboratory-scale low-volume nanometer patterning is presented. The laser-plasma source, which is based on a fluorocarbon liquid-jet target, generates high-brightness lambda = 1.2-1.7 nm x-ray emission with only negligible debris production. The Au/SiNx x-ray mask is fabricated by employing ion milling and a high-contrast e-beam resist. With SAL-601 chemically enhanced resist we demonstrate fabrication of high-aspect-ratio, sub-100 nm structures. The exposure time is currently 20 min using a compact 10 Hz, lambda = 532 nm, 70 mJ/pulse mode-locked Nd:YAG laser. However, the regenerative liquid-jet target is designed for operation with future, e.g., 1000 Hz, lasers resulting in projected exposure times of similar to 10 s. (C) 1997 American Vacuum Society

Covalent immobilization of molecularly imprinted polymer nanoparticles using an epoxy silane.

AbstractMolecularly imprinted polymers (MIPs) can be used as antibody mimics to develop robust chemical sensors. One challenging problem in using MIPs for sensor development is the lack of reliable conjugation chemistry that allows MIPs to be fixed on transducer surface. In this work, we study the use of epoxy silane to immobilize MIP nanoparticles on model transducer surfaces without impairing the function of the immobilized nanoparticles. The MIP nanoparticles with a core–shell structure have selective molecular binding sites in the core and multiple amino groups in the shell. The model transducer surface is functionalized with a self-assembled monolayer of epoxy silane, which reacts with the core–shell MIP particles to enable straightforward immobilization. The whole process is characterized by studying the treated surfaces after each preparation step using atomic force microscopy, scanning electron microscopy, fluorescence microscopy, contact angle measurements and X-ray photoelectron spectroscopy. The microscopy results show that the MIP particles are immobilized uniformly on surface. The photoelectron spectroscopy results further confirm the action of each functionalization step. The molecular selectivity of the MIP-functionalized surface is verified by radioligand binding analysis. The particle immobilization approach described here has a general applicability for constructing selective chemical sensors in different formats

Vascularization of tissue engineered cartilage - Sequential in vivo MRI display functional blood circulation

Establishing functional circulation in bioengineered tissue after implantation is vital for the delivery of oxygen and nutrients to the cells. Native cartilage is avascular and thrives on diffusion, which in turn depends on proximity to circulation. Here, we investigate whether a gridded three-dimensional (3D) bioprinted construct would allow ingrowth of blood vessels and thus prove a functional concept for vascularization of bioengineered tissue. Twenty 10 7 10 7 3-mm 3Dbioprinted nanocellulose constructs containing human nasal chondrocytes or cell-free controls were subcutaneously implanted in 20 nude mice. Over the next 3 months, the mice were sequentially imaged with a 7 T small-animal MRI system, and the diffusion and perfusion parameters were analyzed. The chondrocytes survived and proliferated, and the shape of the constructs was well preserved. The diffusion coefficient was high and well preserved over time. The perfusion and diffusion patterns shown by MRI suggested that blood vessels develop over time in the 3D bioprinted constructs; the vessels were confirmed by histology and immunohistochemistry. We conclude that 3D bioprinted tissue with a gridded structure allows ingrowth of blood vessels and has the potential to be vascularized from the host. This is an essential step to take bioengineered tissue from the bench to clinical practice

Transparent and flexible, nanostructured and mediatorless glucose/oxygen enzymatic fuel cells

Here we detail transparent, flexible, nanostructured, membrane-less and mediator-free glucose/oxygen enzymatic fuel cells, which can be reproducibly fabricated with industrial scale throughput. The electrodes were built on a biocompatible flexible polymer, while nanoimprint lithography was used for their nanostructuring. The electrodes were covered with gold, their surfaces were visualised using scanning electron and atomic force microscopies, and they were also studied spectrophotometrically and electrochemically. The enzymatic fuel cells were fabricated following our previous reports on membrane-less and mediator-free biodevices in which cellobiose dehydrogenase and bilirubin oxidase were used as anodic and cathodic biocatalysts, respectively. The following average characteristics of transparent and flexible biodevices operating in glucose and chloride containing neutral buffers were registered: 0.63 V open-circuit voltage, and 0.6 mu W cm(-2) maximal power density at a cell voltage of 0.35 V. A transparent and flexible enzymatic fuel cell could still deliver at least 0.5 mu W cm(-2) after 12 h of continuous operation. Thus, such biodevices can potentially be used as self-powered biosensors or electric power sources for smart electronic contact lenses. (C) 2015 Elsevier B.V. All rights reserved

Controlled short-linkage assembly of functional nano-objects

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Virus Capsid Dissolution Studied by Microsecond Molecular Dynamics Simulations

Dissolution of many plant viruses is thought to start with swelling of the capsid caused by calcium removal following infection, but no high-resolution structures of swollen capsids exist. Here we have used microsecond all-atom molecular simulations to describe the dynamics of the capsid of satellite tobacco necrosis virus with and without the 92 structural calcium ions. The capsid expanded 2.5% upon removal of the calcium, in good agreement with experimental estimates. The water permeability of the native capsid was similar to that of a phospholipid membrane, but the permeability increased 10-fold after removing the calcium, predominantly between the 2-fold and 3-fold related subunits. The two calcium binding sites close to the icosahedral 3-fold symmetry axis were pivotal in the expansion and capsid-opening process, while the binding site on the 5-fold axis changed little structurally. These findings suggest that the dissociation of the capsid is initiated at the 3-fold axis