Endothelial cell scaffolds generated by 3D direct writing of biodegradable polymer microfibers.

The engineering of large (thickness > 100 µm) tissues requires a microvascular network to supply nutrients and remove waste. To produce microvasculature in vitro, a scaffold is required to mechanically support and stimulate endothelial cell (EC) adhesion and growth. Scaffolds for ECs are currently produced by patterning polymers or other biomaterials into configurations which often possess isotropic morphologies such as porous films and fibrous mats. We propose a new "direct-write" process for fabricating scaffolds composed of suspended polymer microfibers that are precisely oriented in 3D, providing directional architecture for selectively guiding cell growth along a desired pathway. The diameters of the fibers produced with this process were predictably and repeatably controlled through modulation of the system parameters, enabling production of fibers with microvascular-scale diameters (5-20 µm) from a variety of biodegradable polymers. These scaffolds were successfully seeded with ECs, which conformed to the geometry of the fibers and proliferated over the course of one week.

Micro-wilhelmy and related liquid property measurements using constant-diameter nanoneedle-tipped atomic force microscope probes.

The micro-Wilhelmy method is a well-established method of determining surface tension by measuring the force of withdrawing a tens of microns to millimeters in diameter cylindrical wire or fiber from a liquid. A comparison of insertion force to retraction force can also be used to determine the contact angle with the fiber. Given the limited availability of atomic force microscope (AFM) probes that have long constant diameter tips, force-distance (F-D) curves using probes with standard tapered tips have been difficult to relate to surface tension. In this report, constant diameter metal alloy nanowires (referred to as "nanoneedles") between 7.2 and 67 microm in length and 108 and 1006 nm in diameter were grown on AFM probes. F-D and Q damping AFM measurements of wetting and drag forces made with the probes were compared against standard macroscopic models of these forces on slender cylinders to estimate surface tension, contact angle, meniscus height, evaporation rate, and viscosity. The surface tensions for several low molecular weight liquids that were measured with these probes were between -4.2% and +8.3% of standard reported values. Also, the F-D curves show well-defined stair-step events on insertion and retraction from partial wetting liquids, compared to the continuously growing attractive force of standard tapered AFM probe tips. In the AFM used, the stair-step feature in F-D curves was repeatably monitored for at least 0.5 h (depending on the volatility of the liquid), and this feature was then used to evaluate evaporation rates (as low as 0.30 nm/s) through changes in the surface height of the liquid. A nanoneedle with a step change in diameter at a known distance from its end produced two steps in the F-D curve from which the meniscus height was determined. The step features enable meniscus height to be determined from distance between the steps, as an alternative to calculating the height corresponding to the AFM measured values of surface tension and contact angle. All but one of the eight measurements agreed to within 13%. The constant diameter of the nanoneedle also is used to relate viscous damping of the vibrating cantilever to a macroscopic model of Stokes drag on a long cylinder. Expected increases in drag force with insertion depth and viscosity are observed for several glycerol-water solutions. However, an additional damping term (associated with drag of the meniscus on the sidewalls of the nanoneedle) limits the sensitivity of the measurement of drag force for low-viscosity solutions, while low values of Q limit the sensitivity for high-viscosity solutions. Overall, reasonable correspondence is found between the macroscopic models and the measurements with the nanoneedle-tipped probes. Tighter environmental control of the AFM and treatments of needles to give them more ideal surfaces are expected to improve repeatability and make more evident subtle features that currently appear to be present on the F-D and Q damping curves.

Visual force sensing with flexible nanowire buckling springs.

A calibrated method of force sensing is demonstrated in which the buckled shape of a long flexible metallic nanowire, referred to as a 'nanoneedle', is interpreted to determine the applied force. An individual needle of 157 nm diameter by 15.6 µm length is grown on an atomic force microscope (AFM) cantilever with a desired orientation (by the method of Yazdanpanah et al 2005 J. Appl. Phys. 98 073510). Using a nanomanipulator the needle is buckled in the chamber of a scanning electron microscope (SEM) and the buckled shapes are recorded in SEM images. Force is determined as a function of deflection for an assumed elastic modulus by fitting the shapes using the generalized elastica model (De Bona and Zelenika 1997 Proc. Inst. Mech. Eng. C 211 509-17). In this calibration the elastic modulus (68.3 GPa) was determined using an auxiliary AFM measurement, with the needle in the same orientation as in the SEM. Following this calibration the needle was used as a sensor in a different orientation than the AFM coordinates to deflect a suspended PLLA polymer fiber from which the elastic modulus (2.96 GPa) was determined. The practical value of the sensing method does depend on the reliability and ruggedness of the needle. In this study the same needle remained rigidly secured to the AFM cantilever throughout the entire SEM/AFM calibration procedure and the characterization of the nanofiber.

Side-by-side comparison of Raman spectra of anchored and suspended carbon nanomaterials.

Raman spectra of ordered carbon nanomaterials are quite sensitive to surface perturbations, including trace residues, structural defects and residual stress. This is demonstrated by a series of experiments with carbon nanotubes and graphene. Their spectra change due to subtle changes in preparation and attachment to the substrate and to each other. Differences are most clearly seen by forming a material into an air bridge and probing it in the air gap and at the anchor points. A monolayer graphene sheet, shows a larger disorder band at the anchor points than in the air gap. However, a bundle or rope of parallel-aligned single-wall nanotubes shows a larger disorder band in the gap than at the anchor points. For the graphene sheet the substrate surface deforms the graphene, leading to increases in the disorder band. For the rope, the close proximity of the nanotubes to each other appears to produce a larger stress than the rope resting on the substrate.

Antinucleosome antibody-modified liposomes and lipid-core micelles for tumor-targeted delivery of therapeutic and diagnostic agents.

Liposomes and lipid-core micelles prepared of polyethylene glycol-phosphatidylethanolamine (PEG-PE) conjugates have been modified with nucleosome-specific monoclonal antinuclear autoantibody (ANA) 2C5 (mAb 2C5) specifically recognizing a broad variety of cancer cells through the cancer cell surface-bound nucleosomes. mAb 2C5 preserves its specific properties upon the binding with the lipid-based pharmaceutical nanocarriers, and 2C5-modified immunoliposomes and immunomicelles demonstrate an enhanced binding with tumor cells both in vitro and in vivo. We have investigated the delivery of therapeutic and diagnostic agents with such tumor-targeted immunoliposomes and immunomicelles to various tumors in vivo and in vitro. Both lipid-based nanocarriers provided enhanced tumor delivery of imaging agents ((111)In) and antitumor drugs (doxorubicin and photodynamic therapy agents) to tumor cells under different experimental settings. Pharmaceutical lipid-based nanoparticular carriers modified with mAb 2C5 could represent universal systems for tumor-specific delivery of various soluble and insoluble pharmaceuticals.

Oriented nanomaterial air bridges formed from suspended polymer-composite nanofibers.

In a two-step method, carbon nanotubes, inorganic nanowires, or graphene sheets are connected between two anchor points to form nanomaterial air bridges. First, a recently developed method of forming directionally oriented polymer nanofibers by hand-application is used to form suspended composite polymer-nanomaterial fibers. Then, the polymer is sacrificed by thermally induced depolymerization and vaporization, leaving air bridges of the various materials. Composite fibers and bundles of nanotubes as thin as 10 nm that span 1 microm gaps have been formed by this method. Comparable bridges are observed by electrospinning solutions of the same nanomaterial-polymer composites onto micrometer-scale corrugated surfaces. This method for assembling nanomaterial air-bridges provides a convenient way to suspend nanomaterials for mechanical and other property determinations, and for subsequent device fabrication built up from the suspended nanosubstrates.

Paracetamol metabolism in Indian population.

To investigate the profile of paracetamol (CAS 103-90-2, Calpol) metabolism in Indian population and to compare with the profiles of studies conducted in other populations, a study was conducted in 100 healthy male human volunteers. After an overnight fast, the volunteers were administered an oral dose of 1 g of paracetamol, urine was collected up to 8 h and samples were analyzed by high performance liquid chromatography for the estimation of urinary recovery of paracetamol and its metabolites, i.e. sulphate, glucuronide, cysteine and mercapturate conjugates. 25.29 +/- 5.5 (mean +/- S.D.)% of sulphate conjugate, 60.55 +/- 8.5% of glucuronide conjugate, 5.05 +/- 2.1% of unchanged paracetamol, 2.76 +/- 2.4% of cysteine conjugate and 6.37 +/- 3.8% of mercapturate conjugate were recovered. The mean combined recovery of glutathione conjugates (9.13%) in Indians is found to be very high and is equal to that in Caucasians of Scotland (9.3%), which indicates that Indians are equally predisposed to hepatotoxicity as Caucasians.