Niosomal encapsulation of the antitubercular drug, pyrazinamide

It is estimated that more than one-third of the world population is infected with Mycobacterium tuberculosis. Pyrazinamide (PZA) plays a unique role in shortening therapy because it kills a population of semilatent tubercle bacilli residing in an acidic environment. Niosomes are vesicles made up of non-ionic surfactant and exhibit behavior similar to liposomes in vivo. Preparation of PZA niosomes took place using different molar ratios of Span 60 and Span 85, with cholesterol (CH) i.e. Span: CH (1:1) and (4:2). Dicetyl phosphate and stearyl amine were used in preparation of negative and positively charged niosomes, respectively. Free PZA was separated by cooling centrifugation and estimated spectrophotometrically at 268.4 nm. Niosomes were characterized by electron microscopy and differential scanning calorimetry. The highest percentage PZA entrapped was obtained using Span 60 and the molar ratio (4:2:1) negatively charged niosomes. This was followed by the neutral PZA neutral (4:2) Span 60 niosomes. Biological evaluation of selected PZA niosomal formulations took place on guinea pigs infected with M. tuberculosis. The present work is an attempt to target maximum concentration of PZA to the affected site (lungs) and to exclude undesirable side effects and decrease toxicity. Macrophage targeting and overcoming drug resistance is our final goal.     

Transforming the fabrication and biofunctionalization of gold nanoelectrode arrays into versatile electrochemical glucose biosensors

High-density arrays of conducting nanoelectrodes (i.e., nanoelectrode arrays [NEAs]) have been developed on the surface of a single electrode for numerous electrochemical sensing paradigms. However, a scalable fabrication technique and robust biofunctionalization protocol are oftentimes lacking and thus many NEA designs have limited efficacy and overall commercial viability in biosensing applications. In this report, we develop a lithography-free nanofabrication protocol to create large arrays of Au nanoelectrodes on a silicon wafer via a porous anodic alumina template. To demonstrate their effectiveness as electrochemical glucose biosensors, alkanethiol self-assembled monolayers (SAMs) are used to covalently attach the enzyme glucose oxidase to the Au NEA surface for subsequent glucose sensing. The sensitivity and linear sensing range of the biosensor is controlled by introducing higher concentrations of long-chain SAMs (11-mercaptoundecanoic acid: MUA) with short-chain SAMs (3-mercaptopropionic acid: MPA) into the enzyme immobilization scheme. This facile NEA fabrication protocol (that is well-suited for integration into electronic devices) and biosensor performance controllability (via the mixed-length enzyme-conjugated SAMs) transforms the Au NEAs into versatile glucose biosensors. Thus these Au NEAs could potentially be used in important real-word applications such as in health-care and bioenergy where biosensors with very distinct sensing capabilities are needed.     

Microfluidic transendothelial electrical resistance measurement device that enables blood flow and postgrowth experiments

Transendothelial electronic resistance (TEER) measurements are performed across a cell layer immobilized on a microfluidic device that also enables the cell layer to interact with a flowing stream of red blood cells (RBCs). A bipolar pulsed square wave potential is applied across a monolayer of bovine pulmonary artery endothelial cells, and the resulting current response is measured and integrated. The overall impedance of the cell layer provides an indicator of cell layer integrity. After cell seeding on the device, a decrease in TEER signal from 22.3 ± 1.6 μC to 3.5 ± 0.4 μC (corresponding to a resistance of 40.9 ± 2.9 Ω·cm(2) to 259.1 ± 27.4 Ω·cm(2)) was observed after 8 h of cell growth. Intracellular nitric oxide (NO) production by the immobilized endothelial cells that had reached confluence was 34% higher than those cells that had not reached confluence, as indicated by the integrated TEER system. Importantly, this NO production by the confluent endothelium was stimulated by ATP released from RBCs flowing under the endothelial cells. In this construct, the described microfluidic device enables both a TEER-based evaluation of cell layer integrity and molecularly communicated interactions of these cells with a flowing stream of blood components.     

Targeted luminescent near-infrared polymer-nanoprobes for in vivo imaging of tumor hypoxia

Polystyrene nanoparticles (PS-NPs) were doped with an oxygen-sensitive near-infrared (NIR)-emissive palladium meso-tetraphenylporphyrin and an inert reference dye which are both excitable at 635 nm. The nanosensors were characterized with special emphasis on fundamental parameters such as absolute photoluminescence quantum yield and fluorescence lifetime. The PS-NPs were employed for ratiometric dual-wavelength and lifetime-based photoluminescent oxygen sensing. They were efficiently taken up by cultured murine alveolar macrophages, yielding a characteristic and reversible change in ratiometric response with decreasing oxygen concentration. This correlated with the cellular hypoxic status verified by analysis of hypoxia inducible factor-1α (HIF-1α) accumulation. In addition, the surface of PS-NPs was functionalized with polyethylene glycol (PEG) and the monoclonal antibody herceptin, and their binding to HER2/neu-overexpressing tumor cells was confirmed in vitro. First experiments with tumor-bearing mouse revealed a distinctive ratiometric response within the tumor upon hypoxic condition induced by animal sacrifice. These results demonstrate the potential of these referenced NIR nanosensors for in vitro and in vivo imaging that present a new generation of optical probes for oncology.