Interactions between the antifungal drug myclobutanil and gold and silver nanoparticles in Penicillium digitatum investigated by surface-enhanced Raman scattering.

Surface-enhanced Raman scattering (SERS) of an antifungal reagent, myclobutanil (MCB), was performed on Au and Ag nanoparticles (NPs) to estimate the drug-release behaviors in fungal cells. A density functional theory (DFT) calculation was introduced to predict a favorable binding site of MCB to either the Ag or Au atom. Myclobutanil was presumed to bind more strongly to Au than to Ag in their most stable, optimized geometries of the N4 atom in its 1,2,4-triazole unit binding to the metal atom. Strong intensities were observed in the Ag SERS spectra only at acidic pH values, whereas the most prominent peaks in the Au SERS spectra of MCB matched quite well with those of 1,2,4-triazole regardless of pH conditions. The Raman spectral intensities of the MCB-assembled Ag and Au NPs decreased after treatment with either potato dextrose agar (PDA) or glutathione (GSH). Darkfield microscopy and confocal SERS were performed to analyze the MCB-assembled metal NPs inside Penicillium digitatum fungal cells. The results suggested that MCB was released from the metal NPs in the intracellular GSH in the fungi because we observed only fungal cell peaks.

Gold nanorod-assembled PEGylated graphene-oxide nanocomposites for photothermal cancer therapy.

Gold nanorod-attached PEGylated graphene-oxide (AuNR-PEG-GO) nanocomposites were tested for a photothermal platform both in vitro and in vivo. Cytotoxicity of AuNR was reduced after encapsulation with PEG-GO along with the removal of cetyltrimethylammonium bromide (CTAB) from AuNR by HCl treatment. Cellular internalization of the CTAB-eliminated AuNR-PEG-GO nanocomposites was examined using dark-field microscopy (DFM), confocal Raman microscopy and transmission electron microscopy (TEM). To determine the photothermal effect of the AuNR-PEG-GO nanocomposites, A431 epidermoid carcinoma cells were irradiated with Xe-lamp light (60 W cm(-2)) for 5 min after treatment with the AuNR-PEG-GO nanocomposites for 24 h. Cell viability significantly decreased by ~40% when the AuNR-PEG-GO-encapsulated nanocomposites were irradiated with light as compared with the cells treated with only the AuNR-PEG-GO nanocomposites without any illumination. In vivo tumor experiments also indicated that HCl-treated AuNR-PEG-GO nanocomposites might efficiently reduce tumor volumes via photothermal processes. Our graphene and AuNR nanocomposites will be useful for an effective photothermal therapy.

Temperature-dependent structural change of D-penicillamine-capped chiral gold nanoparticles investigated by infrared spectroscopy.

The structure and stability of D-penicillamine-capped gold nanoparticles (d-Pen Au NPs) were studied using spectroscopic tools. The synthesis of d-Pen Au NPs was examined using high-resolution transmission electron microscopy (HR-TEM), UV-vis absorption spectroscopy, and circular dichroism (CD). Temperature-dependent reversible structural changes of d-Pen Au NPs were observed using infrared spectroscopic tools. The three thiol, carboxyl, and amino binding groups of d-Pen were presumed to interact with Au NP surfaces on the basis of the infrared spectral features. d-Pen appeared to form quite a stable structure and desorb at a high temperature above 453 K on Au NPs. Our deconvolution analysis indicated the ν(s)(COO(-)) and ν(as)(COO(-)) carboxylate bands at ∼1,392 and ∼1,560 cm(-1) appeared to be weakened, whereas the amino band at ∼1,595 cm(-1) remained strong in increasing the temperature from 293 to 373 K. On the other hand, the intensities of the zwitter ionic bands at ∼999, ∼1,117, and ∼1,631 cm(-1) for NH(3)(+) appeared to decrease presumably due to the deprotonation process at 373 K. Our infrared spectroscopic study suggests that the deprotonated amino groups bind stronger, whereas the intra-carboxylate bonds become weaker as the temperature increase. Such structural changes of d-Pen Au NPs appeared to be reversible between 293 and 373 K.

Cytotoxicity of serum protein-adsorbed visible-light photocatalytic Ag/AgBr/TiO2 nanoparticles.

Photocytotoxicity of visible-light catalytic Ag/AgBr/TiO(2) nanoparticles (NPs) was examined both in vitro and in vivo. The Ag/AgBr/TiO(2) NPs were prepared by the deposition-precipitation method. Their crystalline structures, atomic compositions, and light absorption property were examined by X-ray diffraction (XRD) patterns, X-ray photoelectron (XPS) intensities, and ultraviolet-visible (UV-vis) diffuse reflectance spectroscopic tools. The Ag/AgBr/TiO(2) NPs appeared to be well internalized in human carcinoma cells as evidenced by transmission electron microscopy (TEM). The cytotoxicity of cetylmethylammonium bromide (CTAB) appeared to be significantly reduced by adsorption of serum proteins in the cellular medium on the NP surfaces. Two types of human cervical HeLa and skin A431 cancer cells were tested to check their viability after the cellular uptake of the Ag/AgBr/TiO(2) NPs and subsequent exposure to an illumination of visible light from a 60 W/cm(2) halogen lamp. Fluorescence images taken to label mitochondria activity suggest that the reactive oxygen species should trigger the photo-destruction of cancer cells. After applying the halogen light illumination for 50-250 min and ∼8 ppm (µg/mL) of photocatalytic Ag/AgBr/TiO(2) NPs, we observed a 40-60% selective decrease of cell viability. Ag/AgBr/TiO(2) NPs were found to eliminate xenograft tumors significantly by irradiating visible light in vivo for 10 min.

Raman detection of localized transferrin-coated gold nanoparticles inside a single cell.

We investigated the cellular uptake behavior of non-fluorescent metal nanoparticles (NPs) by use of surface-enhanced Raman scattering (SERS) combined with dark-field microscopy (DFM). The uptake of Au NPs inside a single cell could also be identified by DFM first and then confirmed by z-depth-dependent SERS at micrometer resolution. Guided by DFM for the location of Au NPs, an intracellular distribution assay was possible using Raman dyes with unique vibrational marker bands in order to identify the three-dimensional location inside the single cell by obtaining specific spectral features. Au NPs modified by 4-mercaptobenzoic acid (MBA) bearing its -COOH surface functional group were used to conjugate transferrin (Tf) protein using the 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC) reaction. The protein conjugation reaction on Au surfaces was examined by means of color change, absorption spectroscopy, and SERS. Our results demonstrate that DFM techniques combined with SERS may have great potential for monitoring biological processes with protein conjugation at the single-cell level.

Infrared spectroscopy characterization of normal and lung cancer cells originated from epithelium

The vibrational spectral differences of normal and lung cancer cells were studied for the development of effective cancer cell screening by means of attenuated total reflection infrared spectroscopy. The phosphate monoester symmetric stretching nus(PO3(2-)) band intensity at ~970 cm-1 and the phosphodiester symmetric stretching nus(PO2-) band intensity at ~1,085 cm-1 in nucleic acids and phospholipids appeared to be significantly strengthened in lung cancer cells with respect to the other vibrational bands compared to normal cells. This finding suggests that more extensive phosphorylation occur in cancer cells. These results demonstrate that lung cancer cells may be prescreened using infrared spectroscopy tools.