Antibacterial nanosized silver substituted hydroxyapatite: synthesis and characterization.

The silver (0.5-3 at %) substituted nanosize hydroxyapatites (AgHAs) were synthesized by microwave processing. The X-ray diffraction (XRD) peaks are very broad, indicating that the AgHAs were of nanosize (30 nm). Transmission electron microscopy analysis shows needle-like morphology of AgHA, having length 60-70 nm and width 15-20 nm. The AgHA phase was stable up to 700 degrees C without any secondary phases. The antibacterial effect of AgHA against Escherichia coli and Staphylococcus aureus was observed by spread plate method, even for low concentration of silver ions (0.5%) with 1 x 10(5) cells/mL of respective bacterial culture, after a 48 h incubation period. However, some colonies of E. coli were seen with a high dose of 1 x 10(8) cells/mL after 24 h. The zone of inhibition by disc diffusion test method was found to vary with the amount of silver in the sintered AgHA pellets, for both the bacteria, after 24 h of inoculation. Osteoblast cell attachment in varying density was noticed on AgHA samples with 0.5, 1.0, and 1.5% silver substitution. However, osteoblast spreading was significantly greater on 0.5% AgHA compared to 1.0 or 1.5% substituted AgHA samples. Thus, the low amount of AgHA has a potential of minimizing the risk of bacterial contamination, without compromising the bioactivity, and is expected to display greater biological efficacy in terms of osseointegration.

Permeability of R6G across Cx43 hemichannels through a novel combination of patch clamp and surface enhanced Raman spectroscopy.

We have measured the permeability of rhodamine-6G across Cx43 hemichannels reconstituted on a pipette tip. Cx43 hemichannels were overexpressed in Sf9 cells, and affinity-purified. The hemichannels were reconstituted in a lipid bilayer on a pipette tip by the tip-dip method. R6G in the pipette permeated across the channels into the bath. The permeability of R6G was quantified by measuring R6G concentration in the bath after several hours by surface enhanced Raman spectroscopy (SERS) with 100 nm silver colloid particles. The ratio of the permeability of dye to salt, as extracted by this combined electrical-SERS technique, is compatible with similar ratios for other dyes across whole gap junction channels. The results for the permeability ratio were further compared to fluorescence measurements. The novel combination of patch and SERS techniques can be extended to quantifying the transport of biologically significant non-fluorescent molecules, such as cAMP and IP3, across 1 nm sized pores, such as the gap junction channel.

Activated pericyte attenuates endothelial functions: nitric oxide-cGMP rescues activated pericyte-associated endothelial dysfunctions.

Hepatic stellate cells are liver-specific pericytes and exist in close proximity with endothelial cells. The activation of liver pericytes is intrinsic to liver pathogenesis, and leads to endothelial dysfunction, including the low bioavailability of nitric oxide (NO). However, the role of nitric oxide in pericyte-endothelium cross-talk has not yet been elucidated. This work examines the cellular mechanism of action of NO in pericyte-mediated endothelial dysfunction. We used in vitro coculture and conditioned medium systems to study the effects of activated liver pericytes on endothelial function, and an egg yolk vascular bed model was used to study the effects of activated pericytes on angiogenesis. This study also demonstrates that activated pericytes attenuate the migration, proliferation, permeability, and NO production of endothelial cells. Our results demonstrate that activated pericytes restrict angiogenesis in egg yolk vascular bed models, and NO supplementation recovers 70% of the inhibition. Our results also demonstrate that supplementation with NO, sildenafil citrate (phosphodiesterase inhibitor), and 8-bromo-cGMP (cGMP analog) partially recovers activated-pericyte-mediated endothelium dysfunction. We conclude that NO-cGMP alleviates activated-pericyte-associated endothelial dysfunction, including angiogenesis, in a cGMP-dependent manner.

Large-area topography analysis and near-field Raman spectroscopy using bent fibre probes.

We present a method for combined far-field Raman imaging, topography analysis and near-field spectroscopy. Surface-enhanced Raman spectra of Rhodamine 6G (R6G) deposited on silver nanoparticles were recorded using a bent fibre aperture-type near-field scanning optical microscope (NSOM) operated in illumination mode. Special measures were taken to enable optical normal-force detection for control of the tip-sample distance. Comparisons between far-field Raman images of R6G-covered Ag particle aggregates with topographic images recorded using atomic force microscopy (AFM) indicate saturation effects due to resonance excitation.