Interaction of erythrocytes with magnetic nanoparticles

Internalization of biocompatible magnetic nanoparticles by red blood cells (RBCs) is a key issue for opportunities of new applications in the biomedical field. In this study, we used in vitro tests to provide evidences of magnetic nanoparticle internalization by mice red blood cells. The internalization process depends upon the nanoparticle concentration and the nanoparticle hydrodynamic radii. The cell internalization of surface-coated maghemite nanoparticles was indirectly tracked by Raman spectroscopy and directly observed using transmission electron microscopy. The observation of nanoparticle cell uptaking using in vitro experiments represents an important breakthrough for the application of nanomagnetism in diagnosis and therapy of RBC-related diseases.     

Synthesis, stability, and cellular internalization of gold nanoparticles containing mixed peptide-poly(ethylene glycol) monolayers

Gold nanoparticles have shown great promise as therapeutics, therapeutic delivery vectors, and intracellular imaging agents. For many biomedical applications, selective cell and nuclear targeting are desirable, and these remain a significant practical challenge in the use of nanoparticles in vivo. This challenge is being addressed by the incorporation of cell-targeting peptides or antibodies onto the nanoparticle surface, modifications that frequently compromise nanoparticle stability in high ionic strength biological media. We describe herein the assembly of poly(ethylene glycol) (PEG) and mixed peptide/PEG monolayers on gold nanoparticle surfaces. The stability of the resulting bioconjugates in high ionic strength media was characterized as a function of nanoparticle size, PEG length, and monolayer composition. In total, three different thiol-modified PEGs (average molecular weight (MW), 900, 1500, and 5000 g mol-1), four particle diameters (10, 20, 30, and 60 nm), and two cell-targeting peptides were explored. We found that nanoparticle stability increased with increasing PEG length, decreasing nanoparticle diameter, and increasing PEG mole fraction. The order of assembly also played a role in nanoparticle stability. Mixed monolayers prepared via the sequential addition of PEG followed by peptide were more stable than particles prepared via simultaneous co-adsorption. Finally, the ability of nanoparticles modified with mixed PEG/RME (RME = receptor-mediated endocytosis) peptide monolayers to target the cytoplasm of HeLa cells was quantified using inductively coupled plasma optical emission spectrometry (ICP-OES). Although it was anticipated that the MW 5000 g mol-1 PEG would sterically block peptides from access to the cell membrane compared to the MW 900 PEG, nanoparticles modified with mixed peptide/PEG 5000 monolayers were internalized as efficiently as nanoparticles containing mixed peptide/PEG 900 monolayers. These studies can provide useful cues in the assembly of stable peptide/gold nanoparticle bioconjugates capable of being internalized into cells.     

In vitro safety toxicology data for evaluation of gold nanoparticles-chronic cytotoxicity, genotoxicity and uptake

Safety and toxic effects of nanoparticles are still largely unexplored due to the multiple aspects that influence their behaviour toward biological systems. Here, we focus the attention on 12 nm spherical gold nanoparticle coated or not with hyaluronic acid compared to its precursor counterpart salt. Results ranging from the effects of a 10-days exposure in an in vitro model with BALB/c 3T3 fibroblast cells show how 12 nm spherical gold nanoparticles are internalized from 3T3 cells by endo-lysosomal pathway by an indirect measurement technique; and how gold nanoparticles, though not being a severe cytotoxicant, induce DNA damage probably through an indirect mechanism due to oxidative stress. While coating them with hyaluronic acid reduces gold nanoparticles cytotoxicity and slows their cell internalization. These results will be of great interest to medicine, since they indicate that gold nanoparticles (with or without coating) are suitable for therapeutic applications due to their tunable cell uptake and low toxicity.     

Can silver nanoparticles be useful as potential biological labels?

Silver (Ag) nanoparticles have unique plasmon-resonant optical scattering properties that are finding use in nanomedical applications such as signal enhancers, optical sensors, and biomarkers. In this study, we examined the chemical and biological properties of Ag nanoparticles of similar sizes, but that differed primarily in their surface chemistry (hydrocarbon versus polysaccharide), in neuroblastoma cells for their potential use as biological labels. We observed strong optical labeling of the cells in a high illumination light microscopy system after 24?h of incubation due to the excitation of plasmon resonance by both types of Ag nanoparticle. Surface binding of both types of Ag nanoparticle to the plasma membrane of the cells was verified with scanning electron microscopy as well as the internalization and localization of the Ag nanoparticles into intracellular vacuoles in thin cell sections with transmission electron microscopy. However, the induction of reactive oxygen species (ROS), degradation of mitochondrial membrane integrity, disruption of the actin cytoskeleton, and reduction in proliferation after stimulation with nerve growth factor were found after incubation with Ag nanoparticles at concentrations of 25?μg?ml(-1) or greater, with a more pronounced effect produced by the hydrocarbon-based Ag nanoparticles in most cases. Therefore, the use of Ag nanoparticles as potential biological labels, even if the surface is chemically modified with a biocompatible material, should be approached with caution.     

Quantitative characterization of gold nanoparticles by field-flow fractionation coupled online with light scattering detection and inductively coupled plasma mass spectrometry

An analytical platform coupling asymmetric flow field-flow fractionation (AF(4)) with multiangle light scattering (MALS), dynamic light scattering (DLS), and inductively coupled plasma mass spectrometry (ICPMS) was established and used for separation and quantitative determination of size and mass concentration of nanoparticles (NPs) in aqueous suspension. Mixtures of three polystyrene (PS) NPs between 20 and 100 nm in diameter and mixtures of three gold (Au) NPs between 10 and 60 nm in diameter were separated by AF(4). The geometric diameters of the separated PS NPs and the hydrodynamic diameters of the Au and PS NPs were determined online by MALS and DLS, respectively. The three separated Au NPs were quantified by ICPMS and recovered at 50-95% of the injected masses, which ranged between approximately 8-80 ng of each nanoparticle size. Au NPs adhering to the membrane in the separation channel was found to be a major cause for incomplete recoveries. The lower limit of detection (LOD) ranged between 0.02 ng Au and 0.4 ng Au, with increasing LOD by increasing nanoparticle diameter. The analytical platform was applied to characterization of Au NPs in livers of rats, which were dosed with 10 nm, 60 nm, or a mixture of 10 and 60 nm nanoparticles by intravenous injection. The homogenized livers were solubilized in tetramethylammonium hydroxide (TMAH), and the recovery of Au NPs from the livers amounted to 86-123% of their total Au content. In spite of successful stabilization with bovine serum albumin even in alkaline medium, separation of the Au NPs by AF(4) was not possible due to association with undissolved remains of the alkali-treated liver tissues as demonstrated by electron microscopy images.     

Thermal Diffusivity Determination of Protoporphyrin IX Solution Mixed with Gold Metallic Nanoparticles

Nanoparticles appear to be ideally suited for applications in targeted thermal effects in medical therapies and photothermally activated drug delivery; all depend critically on the thermal transport between the nanoparticles and the surrounding liquid. In this work thermal lens spectroscopy (TLS) was used to determine the thermal diffusivity of protoporphyrin IX (PpIX) solutions mixed with gold metallic nanoparticles. PpIX disodium salt (DS) was used in a HCl solution at 25%. Fluids containing gold (Au) nanoparticles at different concentrations were prepared and added to the PpIX solutions. For each solution, UV–Vis spectroscopy was used to obtain the optical absorption spectrum, and transmission electron microscopy (TEM) was used to obtain the gold nanoparticle size. From the TLS signal intensity, it was possible to determine the characteristic time constant of the transient thermal by fitting the theoretical expression to the experimental data. From this characteristic time, the thermal diffusivity was obtained for each solution. The results show that the thermal diffusivity of PpIX mixed with gold nanoparticles increases with an increase of the nanoparticle metallic concentration.     

Maximizing DNA loading on a range of gold nanoparticle sizes

We have investigated the variables that influence DNA coverage on gold nanoparticles. The effects of salt concentration, spacer composition, nanoparticle size, and degree of sonication have been evaluated. Maximum loading was obtained by salt aging the nanoparticles to approximately 0.7 M NaCl in the presence of DNA containing a poly(ethylene glycol) spacer. In addition, DNA loading was substantially increased by sonicating the nanoparticles during the surface loading process. Last, nanoparticles up to 250 nm in diameter were found have approximately 2 orders of magnitude higher DNA loading than smaller (13-30 nm) nanoparticles, a consequence of their larger surface area. Stable large particles are attractive for a variety of biodiagnostic assays.     

Gold nanoparticle-modified etched capillaries for open-tubular capillary electrochromatography

The use of gold nanoparticles in conjunction with etched capillary-based open-tubular capillary electrochromatography (OTCEC) to improve the efficiency of separation and the selectivity between selected solutes is described. The fused-silica capillaries (50-microm i.d.) were etched with ammonium hydrogen difluoride, followed by prederivatization of the new surface with (3-mercaptopropyl)trimethoxysilane (MPTMS) for the immobilization of dodecanethiol gold nanoparticles, for OTCEC. The electrochromatography of a "reversed-phase" test mixture and of selected polycylic aromatic hydrocarbons was investigated, and efficient separations and high theoretical plate numbers per meter were obtained. The electroosmotic flow characteristics of the etched gold nanoparticle capillary, unetched gold nanoparticle capillary, bare capillary, and etched bare capillary were studied by varying the percentage of organic modifier in buffer, buffer pH, and separation voltage. Optical microscopy and scanning electron microscopy were used to examine the process of etching and modification and the surface features of the etched gold nanoparticle capillary. The results confirm that dodecanethiol gold nanoparticles bonded on the etched inner wall of the fused-silica capillary can provide sufficient solute-bonded phase interactions to obtain OTCEC separations with reproducible retention, as well as characteristic reversed-phase behavior, even with the inner diameter of the capillary of 50 microm.     

Poly(acrylic acid)-stabilized colloidal gold nanoparticles: synthesis and properties

Combining the intriguing optical properties of gold nanoparticles with the inherent physical and dynamic properties of polymers can give rise to interesting hybrid nanomaterials. In this study, we report the synthesis of poly(acrylic acid) (PAA)-capped gold nanoparticles. The polyelectrolyte-wrapped gold nanoparticles were fully characterized and studied via a combination of techniques, i.e. UV-vis and infrared spectroscopy, dark field optical microscopy, SEM imaging, dynamic light scattering and zeta potential measurements. Although PAA-capped nanoparticles have been previously reported, this study revealed some interesting aspects of the colloidal stability and morphological change of the polymer coating on the nanoparticle surface in an electrolytic environment, at various pH values and at different temperatures.     

Interactions of Skin with Gold Nanoparticles of Different Surface Charge,Shape, and Functionality

The interactions between skin and colloidal gold nanoparticles of different physicochemical characteristics are investigated. By systematically varying the charge, shape, and functionality of gold nanoparticles, the nanoparticle penetration through the different skin layers is assessed. The penetration is evaluated both qualitatively and quantitatively using a variety of complementary techniques. Inductively coupled plasma optical emission spectrometry (ICP‐OES) is used to quantify the total number of particles which penetrate the skin structure. Transmission electron microscopy (TEM) and two photon photoluminescence microscopy (TPPL) on skin cross sections provide a direct visualization of nanoparticle migration within the different skin substructures. These studies reveal that gold nanoparticles functionalized with cell penetrating peptides (CPPs) TAT and R₇ are found in the skin in larger quantities than polyethylene glycol‐functionalized nanoparticles, and are able to enter deep into the skin structure. The systematic studies presented in this work may be of strong interest for developments in transdermal administration of drugs and therapy.     

Material-specific infrared recognition of single sub-10 nm particles by substrate-enhanced scattering-type near-field microscopy

We study the optical material contrast of single nanoparticles in infrared scattering-type near-field optical microscopy (IR s-SNOM) in the presence of strong probe-substrate coupling. It is shown theoretically and experimentally that the contrast depends on both the dielectric properties of the nanoparticles and on their size. We can separate the two dependencies by correlating the simultaneously acquired topography and near-field images pixel-by-pixel. This allows us to establish material-specific mapping of polydisperse nanoparticle mixtures with nanoscale spatial resolution. We experimentally demonstrate the differentiation between sub-10 nm gold and polymer particles adsorbed on a Si substrate. Possible applications of our method range from the material-specific mapping of nanoparticle assemblies to the measurement of the doping concentration in single semiconductor nanoparticles.     

Dendritic functionalization of monolayer-protected gold nanoparticles

This paper describes the facile synthesis of nanoparticle-cored dendrimers (NCDs) and nanoparticle megamers from monolayer-protected gold clusters using either single or multi-step reactions. First, 11-mercaptoundecanoic acid/hexanethiolate-protected gold clusters were synthesized using the Schiffrin reaction followed by the ligand place-exchange reaction. A convergent approach for the synthesis of nanoparticle-cored dendrimers uses a single step reaction that is an ester coupling reaction of hydroxy-functionalized dendrons with carboxylic acid-functionalized gold clusters. A divergent approach, which is based on multi-step reactions, employs the repetition of an amide coupling reaction and a Michael addition reaction to build polyamidoamine dendritic architectures around a nanoparticle core. Nanoparticle megamers, which are large dendrimer-induced nanoparticle aggregates with an average diameter of more than 300 nm, were prepared by the amide coupling reaction between polyamiodoamine [G-2] dendrimers and carboxylic acid-functionalized gold clusters. ¹H NMR spectroscopy, FT-IR spectroscopy, thermogravimetric analysis (TGA), and transmission electron microscopy (TEM) were used for the characterization of these hybrid nanoparticles.     

The effect of nanoparticle on microdomain alignment in block copolymer thin films under an electric field

Controlling microdomain structure of block copolymers (BCPs) under electric field has been one of the most challenging research tasks. In this study, we examined the effect of nanoparticles on the microdomain orientations in BCP/nanoparticle thin films under electric field using cross-sectional transmission electron microscopy experiments. Gold and cadmium selenide nanoparticles with a tailored surface property were incorporated to control microdomain orientations in BCP/nanoparticle thin films by varying dielectric constant of one constituting block. It was revealed that the microdomain orientation of BCP/nanoparticle thin films under electric field was suppressed by the introduction of gold nanoparticles. Thus, it can be inferred that gold nanoparticles can show a shielding effect under external electric field. The effect of complementary parameters such as NPs concentration, exposure time, and field strength were also demonstrated. In addition, it was also found that the suppression effect lessened with cadmium selenide nanoparticles having a dipole from the noncentrosymmetric structure. This work can provide fundamental data for understanding of microdomain alignment behavior of BCP/nanoparticle system under electric field.     

Preparation of gold nanoparticles on silica substrate by radio frequency sputtering

Radio frequency sputtering of gold on amorphous silica substrates was used for the preparation of Au nanoparticles on SiO2. Deposition experiments were carried out in Ar plasmas under mild conditions (RF power = 5/10 W, total pressure = 0.38 mbar, substrate temperature < or = 210 degrees C), focusing in particular on the effect of sputtering time (5/30 min) and substrate temperature on gold nucleation and coalescence, with the aim of obtaining SiO2-supported Au nanoparticles characterized by precise structural and morphological features. To this aim, several analytical techniques were employed for a thorough characterization of the systems properties, including glancing incidence X-ray diffraction, transmission electron microscopy, X-ray photoelectron spectroscopy, atomic force microscopy, and UV-Vis absorption spectroscopy. In particular, the evolution of optical spectra, i.e., of the surface plasmon resonance peak, was used as a probe for the structural features and was related to the results obtained by other characterization techniques. Gold nanoclusters (phi approximately 4/10 nm) dispersed uniformly on silica matrices were obtained under soft conditions, with morphology ranging from island to cluster like. The obtained results make possible a careful modulation of substrate coverage and gold nanoparticle size.     

Quantitative surface acoustic wave detection based on colloidal gold nanoparticles and their bioconjugates

The immobilization scheme of monodispersed gold nanoparticles (10-nm diameter) on piezoelectric substrate surfaces using organosilane molecules as cross-linkers has been developed for lithium niobate (LiNbO3) and silicon oxide (SiO2)/gold-covered lithium tantalate (LiTaO3) of Rayleigh and guided shear horizontal- (guided SH) surface acoustic wave (SAW) sensors. In this study, comparative measurements of gold nanoparticle adsorption kinetics using high-resolution field-emission scanning electron microscopy and SAW sensors allow the frequency responses of SAW sensors to be quantitatively correlated with surface densities of adsorbed nanoparticles. Using this approach, gold nanoparticles are used as the "nanosized mass standards" to scale the mass loading in a wide dynamical range. Rayleigh-SAW and guided SH-SAW sensors are employed here to monitor the surface mass changes on the device surfaces in gas and liquid phases, respectively. The mass sensitivity ( approximately 20 Hz.cm2/ng) of Rayleigh-SAW device (fundamental oscillation frequency of 113.3 MHz in air) is more than 2 orders of magnitude higher than that of conventional 9-MHz quartz crystal microbalance sensors. Furthermore, in situ (aqueous solutions), real-time measurements of adsorption kinetics for both citrate-stabilized gold nanoparticles and DNA-gold nanoparticle conjugates are also demonstrated by guided SH-SAW (fundamental oscillation frequency of 121.3 MHz). By comparing frequency shifts between the adsorption cases of gold nanoparticles and DNA-gold nanoparticle conjugates, the average number of bound oligonucleotides per gold nanoparticle can also be determined. The high mass sensitivity ( approximately 6 Hz.cm2/ng) of guided SH-SAW sensors and successful detection of DNA-gold nanoparticle conjugates paves the way for real-time biosensing in liquids using nanoparticle-enhanced SAW devices.     

Synthesis of dendrimer-stabilized gold-polypyrrole core-shell nanoparticles

Core-shell composite nanoparticles consisting of a gold core and polypyrrole shell were prepared and stabilized with the poly(amidoamine) dendrimer. An in situ redox polymerization technique was used in which pyrrole reduced Au3+ to Au and then oxidized to polypyrrole. The presence of gold nanoparticles as a core was characterized by its surface plasmon absorption peak at 534 nm. Fourier transform infrared spectroscopy confirmed the presence of polypyrrole on the nanoparticle surfaces. The average diameter of the core-shell nanoparticle is 8.7 +/- 1.8 nm with a shell thickness of approximately 1.5-2.0 nm as estimated from the transmission electron microscopy image. Dissolution of the Au core using KCN enabled the formation of hollow polymer nanospheres.     

Electrostatically stabilised nanoparticles: self-organization and electron-beam patterning

The self-organisation of citrate- and magnesium oleate-stabilised gold nanoparticles on SiO2/Si substrates was investigated. In drop deposition, nucleation of citrate-stabilised gold nanoparticles was observed at the rim of the droplet, symmetric or multibranched dendroid gold structures were found in the area between the rim and the central part of the droplet, depending on the drying temperature. Homogeneous submonolayer nanoparticle coverage was obtained by immersion of amineterminated SiO2/Si surfaces into a citrate-stabilised colloidal gold acidic solution. Drop deposition of magnesium oleate-stabilised gold nanoparticles onto the SiO2/Si surfaces resulted in the formation of uniformly close-packed nanoparticle arrays. Under electron beam irradiation, no apparent changes were found for monolayer films of citrate-stabilized particles, but sintering of the nanoparticles was observed in multilayer films. In contrast, coalescence of magnesium oleate-stabilised gold nanoparticle occurred in monolayer films after electron irradiation.     

pH-Dependent gold nanoparticle self-organization on functionalized Si/SiO2 surfaces

The self-organization of citrate- and acrylate-stabilized gold nanoparticles onto SiO₂/hydroxyl-, amino- and nitro-terminated surfaces was investigated as a function of pH. Bare clean Si/SiO₂ substrates were used as the SiO₂/hydroxyl-terminated surfaces and self-assembled monolayers (SAM) of (3-aminopropyl)trimethoxysilane (APTMS) and 3-(4-nitrophenoxy)-propyltrimethoxysilane (NPPTMS) on Si/SiO₂ were employed as the amino- and nitro-terminated surfaces, respectively. All the surfaces were fully characterized by contact angle, atomic force microscopy (AFM), ellipsometry and X-ray photoelectron spectroscopy (XPS). Citrate- and acrylate-stabilized gold nanoparticle stability was also investigated as a function of pH by UV–visible absorption spectroscopy and Z-potentiometry. The gold nanoparticle surface coverage of the substrates was independently estimated by AFM and XPS. The results show that colloid deposition on bare SiO₂/OH surfaces and on NPPTMS monolayers is negligible with the exception of acrylate-stabilized gold nanoparticles which were found to be immobilized on nitro-terminated surfaces at pH lower than 3.5. Nevertheless, APTMS monolayers interact strongly with citrate- and acrylate-stabilized gold nanoparticles exhibiting a dependence of the surface coverage from the pH of the colloidal solution.     

Au-MgF2复合纳米颗粒薄膜的制备和微结构

用射频磁控共溅射法制备了Au体积分数分别为6%、15%、25%、40%、50%和60%的Au-MgF2复合纳米颗粒薄膜.用X射线衍射、透射电镜、X射线光电子能谱对薄膜的微结构和组分进行了测试分析,分析结果表明:制备的Au-MgF2复合纳米颗粒薄膜由fcc-Au晶态纳米微粒镶嵌于主要为非晶态的MgF2陶瓷基体中构成,当Au体积百分含量由15%增至60%时,其平均晶粒尺寸由5.1nm增大到21.2 nm,晶格常数由0.399 84nm增大到0.407 43nm;随Au体积百分含量由6%增至50%,其颗粒平均粒径则由9.8nm增至21.4 nm.名义组分为vol.60%Au-MgF2样品中Au的体积百分含量约为62.6%,与设计值基本一致.     

Quantitative enhanced Raman scattering of labeled DNA from gold and silver nanoparticles

Surface-enhanced resonance Raman scattering (SERRS) from silver nanoparticles using 514.5-nm excitation has been shown to offer huge potential for applications in highly sensitive multiplexed DNA assays. If the technique is to be applied to real biological samples and integrated with other methods, then the use of gold nanoparticles and longer wavelengths of excitation are desirable. The data presented here demonstrate that dye-labeled oligonucleotide sequences can be directly detected by SERRS using gold nanoparticles in a quantitative manner for the first time. The performance of gold and silver nanoparticles as SERRS substrates was assessed using 514.5-, 632.8-, and 785-nm excitation and a range of 13 commercially available dye-labeled oligonucleotides. The quantitative response allowed the limit of detection to be determined for each case and demonstrates that the technique is highly effective, sensitive, and versatile. The possibility of excitation at multiple wavelengths further enhances the multiplexing potential of the technique. The importance of effectively combining the optical properties of the nanoparticle and the dye label is demonstrated. For example, at 632.8-nm excitation, the dye BODIPY TR-X and gold nanoparticles make a strong SERRS combination with very little background fluorescence. This study allows the choice of nanoparticle and dye label for particular experimental setups, and significantly expands the applicability of enhanced Raman scattering for use in many disciplines.