Comparative Study of Catalytic Activity of Recyclable Au/Fe3O4 Microparticles for Reduction Of 2,4-Dinitrophenol and Anionic, Cationic Azo Dyes.

We synthesized Au/Fe3O4 microparticles. Initially, citrate-capped Fe3O4 micro-sized particles were synthesized by the co-precipitation method with an excess amount of trisodium citrate. Gold ions were reduced on the surface of citrate-capped Fe3O4 and grew as gold sub-microparticles with an average diameter of 210 nm on the surface. The characteristic SPR peak of gold nanoparticles on the surface of Fe3O4 was detected at 584 nm, whereas the absorption in the near-infrared region was increased. SEM images has proved that the synthesized Au/Fe3O4 composite microparticles has an average diameter of 1.7 micrometers. The results of XRD patterns proved the existence of both crystal phases of Fe3O4 and Au particles. To investigate the catalytic activity, the reaction rate constant of reduction of 2,4-dinitrophenol (2,4-DNP) and degradation of Congo red (CR), and methylene blue (MB) with NaBH4 in the presence of Au/Fe3O4 catalyst was monitored by UV-Vis spectroscopy. The initial reaction rate constant calculated from the change in characteristic peak absorptions of 2,4-dinitrophenol was 3.97×10-3 s-1, while the reaction rate constants for the degradation of CR and MB were 9.72×10-3 s-1 and 14.25×10-3 s-1 respectively. After 5 cycles, Au/Fe3O4 microparticles preserved 99 % of the reaction rate constant, exhibiting considerable recycling efficiency in the reduction of nitro groups.

Experimental and ab initio studies on the structural, magnetic, photocatalytic, and antibacterial properties of Cu-doped ZnO nanoparticles.

Copper-doped ZnO nanoparticles with a dopant concentration varying from 1-7 mol% were synthesized and their structural, magnetic, and photocatalytic properties were studied using XRD, TEM, SQUID magnetometry, EPR, UV-vis spectroscopy, and first-principles methods within the framework of density functional theory (DFT). Structural analysis indicated highly crystalline Cu-doped ZnO nanoparticles with a hexagonal wurtzite structure, irrespective of the dopant concentration. EDX and EPR studies indicated the incorporation of doped Cu2+ ions in the host ZnO lattice. The photocatalytic activities of the Cu-doped ZnO nanoparticles investigated through the degradation of methylene blue demonstrated an enhancement in photocatalytic activity as the degradation rate changed from 9.89 × 10-4 M min-1 to 4.98 × 10-2 M min-1. By the first-principles method, our results indicated that the Cu(3d) orbital was strongly hybridized with the O(2p) state below the valence band maximum (VBM) due to covalent bonding, and the ground states of the Cu-doped ZnO is favorable for the ferromagnetic state by the asymmetry of majority and minority states due to the presence of unpaired electron.

Energetic stabilities of thiolated pyrimidines on gold nanoparticles investigated by Raman spectroscopy and density functional theory calculations.

The adsorption structures of 2-thiocytosine (2TC) on gold surfaces were examined by means of vibrational Raman spectroscopy and quantum mechanical density functional theory calculations. The 1H-thione-amino form was calculated to be most stable among the six examined tautomers. The three plausible binding geometries of sulfur, pyrimidine nitrogen, and amino group binding modes were calculated to estimate the binding energies of the 1H-thione-amino form with six gold cluster atoms. Thiouracils including 2-thiouracil (2TU), 4-thiouracil (4TU), and 6-methyl-2-thiouracil (6M2TU) were also studied to compare their relative binding energies on gold atoms. The intracellular localization of a DNA base analog of 2TC on gold nanoparticles (AuNPs) in HeLa cells was identified by means of surface-enhanced Raman scattering. AuNPs were modified with 2TC by self-assembly. Our dark-field microscopy and z-depth-dependent confocal Raman spectroscopy indicated that 2TC-assembled AuNPs could be found inside cancer cells. On the other hand, we did not observe noticeably strong Raman peaks in the cases of thiouracils including 2TU, 4TU, and 6M2TU. This may be due to the additional amino group of 2TC, which can lead to a stronger binding of adsorbates on AuNPs.

Nonidentical intracellular drug release rates in Raman and fluorescence spectroscopic determination.

Intracellular drug release rates were measured by monitoring mitoxantrone (MTX) on gold nanoparticle (AuNP) carriers by means of real-time label-free bimodal imaging with confocal Raman and fluorescence spectroscopy. The quenching nature of the MTX-AuNPs by nanometal surface energy transfer (NSET) was analyzed using the determined Stern-Volmer constant of KSV = 2.28 × 10(9) M(-1). The amount of MTX released was estimated by both the decrease in the surface-enhanced resonance Raman scattering (SERRS) signal and the increase in the fluorescence intensity. Both SERRS and NSET provide quantitative relationships between the spectral intensities of MTX concentrations in solution. Inside live cells, the signal decay profiles of the drug release from AuNPs appeared to be faster at the beginning of the bond-breaking drug release for the SERRS (R(-12)) than the recovery time of the NSET (R(-4) or R(-6)). In the first 45 min, a rather fast decay rate k of 0.0252 min(-1) with a short half-life t1/2 of 27.5 min was observed, whereas the rate became significantly slower in a diffusion process, 0.0093 min(-1) with a longer half-life of 101.4 min, after 45 min.

Colloidal gold nanoparticle conjugates of gefitinib.

Gefitinib (GF) is a US Food and Drug Administration-approved epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor for treating the lung cancers. We fabricated colloidal gold nanoparticle (AuNP) conjugates of the GF anticancer drug by self-assembly to test their potency against A549, NCI-H460, and NCI-H1975 lung cancer cells. GF adsorption on AuNP surfaces was examined by UV-vis absorption spectra and surface-enhanced Raman scattering. Density functional theory calculations were performed to estimate the energetic stabilities of the drug-AuNP composites. The N1 nitrogen atom of the quinazoline ring of GF was calculated to be more stable than the N3 in binding Au cluster atoms. The internalizations of GF-coated AuNPs were examined by transmission electron and dark-field microscopy. A cell viability test of AuNP-GF conjugates with the EGFR antibody exhibited much higher reductions than free GF for A549, NCI-H460, and NCI-H1975 lung cancer cells after treatment for 48.

Adsorption and desorption of tyrosine kinase inhibitor erlotinib on gold nanoparticles.

We investigated interfacial behaviors of erlotinib (EL) on gold nanoparticles (AuNPs) by means of Raman spectroscopy. The adsorption reactions and structures of EL on AuNP surfaces were examined by UV-Vis absorption spectroscopy and surface-enhanced Raman scattering (SERS). Density functional theory calculations were performed to estimate the energetic stabilities of the drug-AuNP composites. Among the binding units in EL, the acetylenic C≡C group was calculated to be the most strongly binding on the AuNP cluster atoms, consistent with the SERS spectra. The concentration-dependent SERS spectra indicated that ∼10(-5) M of EL exhibited the highest SERS signals. The attached EL appeared to desorb more efficiently with 2mM glutathione than with cell culture media. The lack of a strong SERS signal of EL in the dark-field microscopy images of AuNP-EL complexes suggested almost complete desorption of EL inside cells.

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 nanoparticle silica nanopeapods.

A subnanometer gap-separated linear chain gold nanoparticle (AuNP) silica nanotube peapod (SNTP) was fabricated by self-assembly. The geometrical configurations of the AuNPs inside the SNTPs were managed in order to pose either a single-line or a double-line nanostructure by controlling the diameters of the AuNPs and the orifice in the silica nanotubes (SNTs). The AuNPs were internalized and self-assembled linearly inside the SNTs by capillary force using a repeated wet-dry process on a rocking plate. Transmission electron microscopy (TEM) images clearly indicated that numerous nanogap junctions with sub-1-nm distances were formed among AuNPs inside SNTs. Finite-dimension time domain (FDTD) calculations were performed to estimate the electric field enhancements. Polarization-dependent surface-enhanced Raman scattering (SERS) spectra of bifunctional aromatic linker p-mercaptobenzoic acid (p-MBA)-coated AuNP-embedded SNTs supported the linearly aligned nanogaps. We could demonstrate a silica wall-protected nanopeapod sensor with single nanotube sensitivity. SNTPs have potential application to intracellular pH sensors after endocytosis in mammalian cells for practical purposes. The TEM images indicated that the nanogaps were preserved inside the cellular constituents. SNTPs exhibited superior quality SERS spectra in vivo due to well-sustained nanogap junctions inside the SNTs, when compared to simply using AuNPs without any silica encapsulation. By using these SNTPs, a robust intracellular optical pH sensor could be developed with the advantage of the sustained nanogaps, due to silica wall-protection.

Detection of the mycotoxin citrinin using silver substrates and Raman spectroscopy.

We detected a trace amount of the mycotoxin citrinin using surface-enhanced Raman scattering (SERS) on silver nanoparticle (Ag NP) surfaces. The SERS substrate on hydrophobic Teflon films was also introduced to observe the citrinin peaks. A broad band at ∼1382cm(-1), which was ascribed to the symmetric carboxylate stretching mode, was observed in addition to an antisymmetric carboxylate stretching mode at ∼1568cm(-1) in the Raman spectra. The spectral feature indicated that citrinin would adsorb on Ag NPs via its carboxylate form. Based on density functional theory (DFT) calculations, vibrational mode analysis was performed to compare the Raman spectra of citrinin. DFT calculations also predicted that a bidentate bridge configuration through O15 and O16 atoms in citrinin would be the most stable on three Ag atoms. After treating with Ag NPs, observation of citrinin peaks was attempted in fungal cells of Penicillium citrinum. This work may provide useful insights into the direct observation of the hazardous citrinin mycotoxin using SERS by understanding its adsorption behaviors on Ag surfaces.

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.

Label-free Raman spectroscopy for accessing intracellular anticancer drug release on gold nanoparticles.

We investigated glutathione (GSH)-induced purine or pyrimidine anticancer drug release on gold nanoparticle (AuNP) surfaces by means of label-free Raman spectroscopy. GSH-triggered releases of 6-thioguanine (6TG), gemcitabine (GEM), acycloguanosine (ACY), and fadrozole (FAD) were examined in a comparative way by means of surface-enhanced Raman scattering (SERS). The GSH-induced dissociation constant of GEM (or ACY/FAD) from AuNPs was estimated to be larger by more than 38 times than that of 6TG from the kinetic relationship. Tripeptide control experiments were presented to check the turn-off Raman signalling mechanism. Dark-field microscopy (DFM) and transmission electron microscopy (TEM) indicated the intracellular AuNP loads. After their cellular uptake, GEM, ACY, and FAD would not show SERS intensities as strong as 6TG. This may be due to easier release of GEM, ACY, and FAD than 6TG by intracellular reducing species including GSH. We observed fairly strong SERS signals of GEM and 6TG in cell culture media solution. Our CCK-8 cytotoxicity assay data support that 6TG-AuNPs did not exhibit a substantial decrease in cell viability presumably due to strong binding. Label-free confocal Raman spectroscopy can be utilized as an effective tool to access intracellular anticancer drug release.

Confocal Raman microspectroscopic study of folate receptor-targeted delivery of 6-mercaptopurine-embedded gold nanoparticles in a single cell.

We investigate the cellular uptake behaviors and efficacy of folate-coated gold nanoparticles (AuNPs) for the targeted drug delivery system in human cancer cells. Folate-conjugated AuNPs embedded with a purine analogue cancer drug of 6-mercaptopurine (6MP) were assembled via a 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride (EDC) coupling reaction between the amino group of 4-aminobenzenethiol (ABT) and the carboxyl group of folic acid. The assembly of folate and 6MP on AuNPs has been examined by absorption spectroscopy, transmission electron microscopy (TEM), and confocal Raman spectroscopy. The internalization of the conjugated AuNPs inside the folate receptor-positive HeLa and KB cells was checked by TEM and dark-field microscopy (DFM) combined with label-free confocal spectroscopy over the depth variable z at a micrometer resolution. DFM live cell imaging of folate-conjugated AuNPs in HeLa cells indicated that the targeted AuNPs appeared to attach on the cell surfaces and enter into the cell with an hour. The cell viability was also compared to estimate the efficacy of folate-conjugated AuNP delivery systems. Folate receptor-targeted AuNP systems appeared to decrease cancer cell viability both in vitro and in vivo more than did the use of the 6MP-coated AuNPs drug without any targeting systems.

Aggregation effects of gold nanoparticles for single-base mismatch detection in influenza A (H1N1) DNA sequences using fluorescence and Raman measurements.

Aggregation effects of gold nanoparticles (AuNPs) were examined for the discrimination of single point mutations through the hybridization of oligonucleotides (25-50 nM) modified with a fluorescent Texas red dye. The sequences of oligonucleotides were designed to detect the H1N1 virus gene. Single-base mismatch detection due to different adsorption propensities of oligonucleotides could be achieved using fluorescence quenching and surface-enhanced Raman scattering (SERS) properties of the dye. We observed that the addition of perfectly matched double stranded DNA (pmdsDNA), modified with the Texas red dye in the suspension of citrate-reduced AuNPs could increase fluorescence recovery intensities more substantially than either single-base mismatched double stranded DNA (sbmdsDNA) or single stranded DNA (ssDNA). We also tested DNA hybridization under both aggregation and near non-aggregation conditions for fluorescence measurements. A spectral difference in fluorescence intensity between pmdsDNA and sbmdsDNA appeared to be more discriminating under near non-aggregation than aggregation conditions. On the other hand, the SERS intensities of pmdsDNA and sbmdsDNA decreased more significantly than that of ssDNA under aggregation conditions, whereas we could not observe any SERS intensities under non-aggregation conditions.

Real-time monitoring of glutathione-triggered thiopurine anticancer drug release in live cells investigated by surface-enhanced Raman scattering.

We investigated in vitro and in vivo glutathione (GSH)-induced intracellular thiopurine anticancer drug release on gold nanoparticle (Au NP) surfaces by means of label-free confocal Raman spectroscopy. Direct monitoring of GSH-triggered release of 6-mercaptopurine (6MP) and 6-thioguanine (6TG) was achieved in real time. Live cell imaging technique provides a nanomolar range release of 6MP and 6TG from Au NP surfaces after the injection of external GSH. In vivo SERS spectra of 6TG were obtained from the subcutaneous sites in living mice after GSH treatment. GSH-triggered releases of Cy5-dye assembled on 6TG-capped Au NPs were also compared using independent fluorescence measurements. Our work demonstrates that the time-lapse Raman spectroscopic tools are useful for monitoring of the controlled release of thiopurine drug molecules in vitro and in vivo.

Preferential adsorption of fetal bovine serum on bare and aromatic thiol-functionalized gold surfaces in cell culture media.

Intracellular uptake of serum-coated gold nanoparticles (AuNPs) in a single mammalian cell was examined in order to investigate the interactions of cell culture media and aromatic thiol-functionalized gold surfaces using micro-spectroscopic tools. The AuNPs modified by the aromatic thiols of para-aminobenzenethiol (ABT), para-hydroxy benzenethiol (HBT), and para-carboxylic benzenethiol (CBT, para-mercaptobenzoic acid) bearing NH(2), OH, and COOH surface functional groups are presumed to adsorb the serum proteins as indicated from the compiled quartz crystal microbalance (QCM) data. The QCM results indicate that among the constituents, fetal bovine serum (FBS) should be the major adsorbate species on AuNPs incubated in Roswell Park Memorial Institute (RPMI) medium. The functionalized AuNPs were found to be internalized as an aggregation state in mammalian cells as evidenced by transmission electron microscopy (TEM) images. We monitored such cellular uptake behaviors of aromatic thiol-modified AuNPs using dark-field microscopy (DFM)-guided confocal surface-enhanced Raman scattering techniques in order to identify the three-dimensional localization inside the single cell. We found that the uptake amounts of ABT, HBT, and CBT were similar by counting up to 70 particles inside the cells incubated in the solution mixture of the aromatic thiol and 1,4-phenylenediisocyanide (PDIC) as a reference. This result indicates for the short aromatic thiol compounds, the AuNPs should enter the cell after the serum-coating regardless of the surface functional groups. Considering that the aromatic thiols have little effect on the serum coating, the DFM/SERS method is an effective tool for monitoring the localization of AuNPs inside a single cell.

Interfacial behavior of benzoic acid and phenylphosphonic acid on nanocrystalline TiO(2) surfaces.

The interfacial behavior of self-assembled thin films of benzoic acid (BA) and phenylphosphonic acid (PPOA) anchored on TiO(2) surfaces was studied by using temperature-dependent diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy. On the basis of the disappearance of the OH band from the infrared spectra at room temperature, BA and PPOA appear to adsorb onto TiO(2) surfaces through carboxylate and phosphonate groups, respectively. Above 420 degrees C, DRIFT spectra indicated that both BA and PPOA desorb from TiO(2) surfaces; however, dissimilar desorption behavior could be inferred for BA and PPOA from their temperature-dependent spectral changes. The benzene ring modes of PPOA remained above 420 degrees C, whereas those of BA disappeared. Density functional theory calculations showed that the adsorption of BA and PPOA on TiO(2) surfaces corresponded to bidentate bridging geometry on TiO(2) surfaces, and the adsorption of PPOA is stronger than that of BA. The monodentate structures with energy differences of 4.9 and 9.1 kcal mol(-1) from the most stable bidentate structures of BA and PPOA, respectively, from the DFT calculations appeared to be possible, particularly at the high temperatures above 420 degrees C, as indicated by the intensified OH bands. The geometry of PPOA was also estimated to be more upright standing than that of BA on TiO(2) surfaces, which may lead a rather straight detachment from the TiO(2) surfaces based on the presence of in-plane ring modes in the DRIFT spectra at the higher temperature.

Hydrogen bonding-induced color recovery of gold nanoparticles upon conjugation of amino acids.

Hydrogen bonding-induced redispersion of the aggregated Au nanoparticles upon N-hydroxysuccinimide ester bioconjugations may provide a simple and colorimetric tool as an optical sensor to detect a trace amount of amino acids as low as approximately 10(-6) M in an aqueous solution.

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 nu(s)(PO3(2-)) band intensity at ~970 cm(-1) and the phosphodiester symmetric stretching nu(s)(PO2(-)) band intensity at approximately 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.

Adsorption changes of cyclohexyl isothiocyanate on gold surfaces.

The adsorption and structure of cyclohexyl isothiocyanate (CHIT) on gold surfaces has been investigated by surface-enhanced Raman scattering (SERS) and scanning tunneling microscopy (STM). Depending on the concentration, the spectral changes of the NCS stretching vibration on gold nanoparticles appeared to be more conspicuous than those of cyclohexyl ring modes. Both equatorial and axial chair conformers of CHIT were found to exist at low bulk concentrations near the monolayer coverage limit, whereas the equatorial chair conformer appeared to be dominant at high bulk concentrations. It was also observed that the ring conversion of equatorial to axial conformers can easily occur at higher temperatures, and the mole fraction of the axial form is assumed to increase with increasing temperature from 30 to 60 degrees C. Alternatively, STM imaging revealed that the adsorption of CHIT molecules on Au(1 1 1) leads to the formation of disordered SAMs with a few vacancy islands, as opposed to the formation of well-ordered self-assembled monolayers (SAMs) by cyclohexanethiols.