Fibrous polymer nanomaterials for biomedical applications and their transport by fluids: an overview.

Over the past few decades, there has been strong interest in the development of new micro- and nanomaterials for biomedical applications. Their use in the form of capsules, particles or filaments suspended in body fluids is associated with conformational changes and hydrodynamic interactions responsible for their transport. The dynamics of fibres or other long objects in Poiseuille flow is one of the fundamental problems in a variety of biomedical contexts, such as mobility of proteins, dynamics of DNA or other biological polymers, cell movement, tissue engineering, and drug delivery. In this review, we discuss several important applications of micro and nanoobjects in this field and try to understand the problems of their transport in flow resulting from material-environment interactions in typical, crowded, and complex biological fluids. Our aim is to elucidate the relationship between the nano- and microscopic structures of elongated polymer particles and their flow properties, thus opening the possibility to design nanoobjects that can be efficiently transported by body fluids for targeted drug release or local tissue regeneration.

Structural Analysis of Collapsed, and Twisted and Collapsed, Multiwalled Carbon Nanotubes by Atomic Force Microscopy.

A fully collapsed multiwalled carbon nanotube (MWCNT1) section and a different twisted and fully collapsed MWCNT were observed with tapping-mode atomic force microscopy. The collapsed section of MWCNT1 was significantly more flexible than the uncollapsed sections, and advanced 120 nm within 1 month. The collapse of MWCNT1 was most likely initiated by its interaction with the surface, and possibly a water meniscus. The ability of carbon nanotubes to radially deform under the influence of surface interactions is in striking contrast with their extremely high axial rigidity.

Investigation of the radial deformability of individual carbon nanotubes under controlled indentation force

Tapping-mode atomic force microscopy was used to study the radial deformability of a multiwalled carbon nanotube (MWCNT). By imaging the MWCNT under different tapping forces, we were able to demonstrate its remarkable reversible radial deformability (up to approximately 40%) and reveal internal discontinuities along its length. The values of the effective elastic modulus of several sections of the MWCNT in the radial direction were estimated with the Hertz model.

Electrospinning of poly(lactic acid)/polyhedral oligomeric silsesquioxane nanocomposites and their potential in chondrogenic tissue regeneration.

The study was conducted to evaluate the cytocompatibility and hydrolytic degradability of the new poly(lactic acid)/polyethylene glycol-polyhedral oligomeric silsesquioxane (peg-POSS/PLLA) nanocomposite as potential material for cartilage regeneration. PLLA scaffolds containing 0 to 5% of peg-POSS were fabricated by electrospinning. Human mesenchymal stem cells (hMSC's) were cultured in vitro to evaluate the cytocompatibility of the new nanocomposite material. Hydrolytic degradation studies were also carried out to analyze the mass loss rate of the nanocomposites through time. The addition of the peg-POSS to the PLLA did not affect the processability of the nanocomposite by electrospinning. It was also observed that peg-POSS did not show any relevant change in fibers morphology, concluding that it was well dispersed. However, addition of peg-POSS caused noticeable decrease in mean fiber diameter, which made the specific surface area of the scaffold to rise. hMSC's were able to attach, to proliferate, and to differentiate into chondrocytes in a similar way onto the different types of electrospun peg-POSS/PLLA and pure PLLA scaffolds, showing that the peg-POSS as nano-additive does not exhibit any cytotoxicity. The hydrolytic degradation rate of the material was lower when peg-POSS was added, showing a higher durability of the nanocomposites through time. Results demonstrate that the addition of peg-POSS to the PLLA scaffolds does not affect its cytocompatibility to obtain hyaline cartilage from hMSC's.

Effect of self-assembled monolayers on charge injection and transport in poly(3-hexylthiophene)-based field-effect transistors at different channel length scales.

Charge injection and transport in bottom-contact regioregular-poly(3-hexylthiophene) (rr-P3HT) based field-effect transistors (FETs), wherein the Au source and drain contacts are modified by self-assembled monolayers (SAMs), is reported at different channel length scales. Ultraviolet photoelectron spectroscopy is used to measure the change in metal work function upon treatment with four SAMs consisting of thiol-adsorbates of different chemical composition. Treatment of FETs with electron-poor (electron-rich) SAMs resulted in an increase (decrease) in contact metal work function because of the electron-withdrawing (-donating) tendency of the polar molecules. The change in metal work function affects charge injection and is reflected in the form of the modulation of the contact resistance, R(C). For example, R(C) decreased to 0.18 MΩ in the case of the (electron-poor) 3,5-bis-trifluoromethylbenzenethiol treated contacts from the value of 0.61 MΩ measured in the case of clean Au-contacts, whereas it increased to 0.97 MΩ in the case of the (electron-rich) 3-thiomethylthiophene treated contacts. Field-effect mobility values are observed to be affected in short-channel devices (<20 µm) but not in long-channel devices. This channel-length-dependent behavior of mobility is attributed to grain-boundary limited charge transport at longer channel lengths in these devices.

In situ atomic force microscopy study of Alzheimer's beta-amyloid peptide on different substrates: new insights into mechanism of beta-sheet formation.

We have applied in situ atomic force microscopy to directly observe the aggregation of Alzheimer's beta-amyloid peptide (Abeta) in contact with two model solid surfaces: hydrophilic mica and hydrophobic graphite. The time course of aggregation was followed by continuous imaging of surfaces remaining in contact with 10-500 microM solutions of Abeta in PBS (pH 7.4). Visualization of fragile nanoscale aggregates of Abeta was made possible by the application of a tapping mode of imaging, which minimizes the lateral forces between the probe tip and the sample. The size and the shape of Abeta aggregates, as well as the kinetics of their formation, exhibited pronounced dependence on the physicochemical nature of the surface. On hydrophilic mica, Abeta formed particulate, pseudomicellar aggregates, which at higher Abeta concentration had the tendency to form linear assemblies, reminiscent of protofibrillar species described recently in the literature. In contrast, on hydrophobic graphite Abeta formed uniform, elongated sheets. The dimensions of those sheets were consistent with the dimensions of beta-sheets with extended peptide chains perpendicular to the long axis of the aggregate. The sheets of Abeta were oriented along three directions at 120 degrees to each other, resembling the crystallographic symmetry of a graphite surface. Such substrate-templated self-assembly may be the distinguishing feature of beta-sheets in comparison with alpha-helices. These studies show that in situ atomic force microscopy enables direct assessment of amyloid aggregation in physiological fluids and suggest that Abeta fibril formation may be driven by interactions at the interface of aqueous solutions and hydrophobic substrates, as occurs in membranes and lipoprotein particles in vivo.

Characterization of two forms of cadmium phosphide by magic-angle spinning 31P NMR.

Annealing of commercial grade cadmium phosphide (CD3P2) at 600 K produces a material which, in magic-angle spinning spin-lattice 31P NMR relaxation experiments, has broad lines and multiple T1 values. By contrast, sublimation at 900 K results in a crystalline material with narrow lines and a single T1. However, both materials have the same Cd-P lattice spacings as determined by rotational-echo, double-resonance 31P NMR with 113Cd dephasing. Both materials also have closely similar X-ray diffraction powder patterns. These results are interpreted in terms of a distribution of lattice vacancies in the annealed material, creating structural heterogeneity but with no substantial change in lattice parameters from those of the sublimed cadmium phosphide.

Interaction of DNA-dependent protein kinase with DNA and with Ku: biochemical and atomic-force microscopy studies.

DNA-dependent protein kinase (DNA-PK or the scid factor) and Ku are critical for DNA end-joining in V(D)J recombination and in general non-homologous double-strand break repair. One model for the function of DNA-PK is that it forms a complex with Ku70/86, and this complex then binds to DNA ends, with Ku serving as the DNA-binding subunit. We find that DNA-PK can itself bind to linear DNA fragments ranging in size from 18 to 841 bp double-stranded (ds) DNA, as indicated by: (i) mobility shifts; (ii) crosslinking between the DNA and DNA-PK; and (iii) atomic-force microscopy. Binding of the 18 bp ds DNA to DNA-PK activates it for phosphorylation of protein targets, and this level of activation is not increased by addition of purified Ku70/86. Ku can stimulate DNA-PK activity beyond this level only when the DNA fragments are long enough for the independent binding to the DNA of both DNA-PK and Ku. Atomic-force microscopy indicates that under such conditions, the DNA-PK binds at the DNA termini, and Ku70/86 assumes a position along the ds DNA that is adjacent to the DNA-PK.

Estimation of acoustical streaming: theoretical model, Doppler measurements and optical visualisation.

An approximate solution for the streaming velocity generated by flat and weakly focused transducers was derived by directly solving the Dirichlet boundary conditions for the Poisson equation, the solution of the Navier-Stokes equation for the axial components of the streaming velocity. The theoretical model was verified experimentally using a 32 MHz pulsed Doppler unit. The experimental acoustical fields were produced by three different 4 mm diameter flat and focused transducers driven by the transmitter generating the average acoustic power within the range from 1 microW to 6 mW. The streaming velocity was measured along the ultrasonic beam from 0 to 2 cm. Streaming was induced in a solution of water and corn starch. The experimental results showed that for a given acoustic power the streaming velocity was independent of the starch density in water, changed from 0.3 to 40 grams of starch in 1 l of distilled water. For applied acoustic powers, the streaming velocity changed linearly from 0.2 to 40 mm/s. Both, the theoretical solutions for plane and focused waves and the experimental results were in good agreement. The streaming velocity field was also visualised using the particle image velocimetry (PIV) and two different evaluation methods. The first based on the FFT-based cross-correlation analysis between small sections for each pair of images and the second employing the algorithm of searching for local displacements between several images.

Complex nanostructured materials from segmented copolymers prepared by ATRP.

The development of new controlled/living radical polymerization processes, such as Atom Transfer Radical Polymerization (ATRP) and other techniques such as nitroxide mediated polymerization and degenerative transfer processes, including RAFT, opened the way to the use of radical polymerization for the synthesis of well-defined, complex functional nanostructures. The development of such nanostructures is primarily dependent on self-assembly of well-defined segmented copolymers. This article describes the fundamentals of ATRP, relevant to the synthesis of such systems. The self-assembly of block copolymers prepared by ATRP is illustrated by three examples. In the first, block copolymers of poly(butyl acrylate) with polyacrylonitrile phase separate, leading to spherical, cylindrical or lamellar morphologies, depending on the block copolymer composition. At a higher temperature, polyacrylonitrile block converts to nanostructured carbon clusters, whereas poly(butyl acrylate) block serves as a sacrificial block, aiding the development of designed nanostructures. In the second example, conductive nanoribbons of poly(n-hexylthiophene) surrounded by a matrix of organic polymers are formed from block copolymers prepared by ATRP. The third example describes an inorganic-organic hybrid system consisting of hard nanocolloidal silica particles (approximately 20 nm) grafted by ATRP with well-defined polystyrene-poly(benzyl acrylate) block copolymer chains (approximately 1000 chains per particle). Silica cores in this system are surrounded by a rigid polystyrene inner shell and softer polyacrylate outer shell.

Face, content and construct validity of the University of Washington virtual reality transurethral prostate resection trainer.

PURPOSE: We examined the face, content and construct validity of version 1.0 of the University of Washington transurethral prostate resection (TURP) trainer. MATERIALS AND METHODS: Version 1.0 of a virtual reality based simulator for transurethral skills was developed at our laboratory by integrating TURP hardware with our virtual 3-dimensional anatomy, irrigation control, cutting, bleeding and haptics force feedback. A total of 72 board certified urologists and 19 novices completed a pre-task questionnaire, viewed an introductory training video and performed a pre-compiled 5-minute resection task. The simulator logged operative errors, gm resected, blood loss, irrigant volume, foot pedal use and differential time spent with orientation, cutting or coagulation. Trainees and experts evaluated the simulator on a modified likert scale. The 2-tailed Levene t test was used to compare means between experts and novices. RESULTS: Overall version 1.0 content was between slightly and moderately acceptable. Experts spent less time with orientation (p < 0.0001), resected more total tissue (p < 0.0001), had more gm resected per cut (p = 0.002) and less blood loss per gm resected (p = 0.032), used less irrigant per gm resected (p = 0.02) and performed fewer errors (p < 0.0001) than novices. CONCLUSIONS: We established the face, content and construct validity for version 1.0 of the University of Washington TURP trainer to simulate the skills necessary to perform TURP. A predictive validity study showing a translation of skills from the virtual environment to the operating room will complete the validation of this model.

Packaging of DNA by shell crosslinked nanoparticles.

We demonstrate compaction of DNA with nanoscale biomimetic constructs which are robust synthetic analogs of globular proteins. These constructs are approximately 15 nm in diameter, shell crosslinked knedel-like (SCKs) nanoparticles, which are prepared by covalent stabilization of amphiphilic di-block co-polymer micelles, self-assembled in an aqueous solution. This synthetic approach yields size-controlled nanoparticles of persistent shape and containing positively charged functional groups at and near the particle surface. Such properties allow SCKs to bind with DNA through electrostatic interactions and facilitate reduction of the DNA hydrodynamic diameter through reversible compaction. Compaction of DNA by SCKs was evident in dynamic light scattering experiments and was directly observed by in situ atomic force microscopy. Moreover, enzymatic digestion of the DNA plasmid (pBR322, 4361 bp) by Eco RI was inhibited at low SCK:DNA ratios and prevented when [le]60 DNA bp were bound per SCK. Digestion by Msp I in the presence of SCKs resulted in longer DNA fragments, indicating that not all enzyme cleavage sites were accessible within the DNA/SCK aggregates. These results have implications for the development of vehicles for successful gene therapy applications.