Bio-synthesis of gold nanoparticles by human epithelial cells, in vivo

Healthy epithelial cells, in vivo, have the ability to synthesize gold nanoparticles when aqueous tetrachloroauric acid is made to react with human skin. Neither a reducing agent nor a protecting chemical is needed for this bio-synthesis method. The first indication of gold nanoparticle formation is the staining of the skin, which turns deep purple. Stereoscopic optical micrographs of human skin tissue in contact with aqueous tetrachloroauric acid clearly show the staining of the epithelial cells. The UV-Vis spectrum of these epithelial cells shows an absorption band with a maximum at 553 nm. This absorption peak is within the wavelength region where the surface plasmon resonance (SPR) band of aqueous colloidal gold exhibits a maximum. Transmission electron micrographs show that gold nanoparticles synthesized by epithelial cells have sizes between 1 and 100 nm. The electron diffraction pattern of these nanoparticles reveals a crystalline structure whose interplanar distances correspond to fcc metallic gold. Transmission electron micrographs of ultra-thin (70 nm thick) slices of epithelial cells clearly and undoubtedly demonstrate that gold nanoparticles are inside the cell. According to high resolution transmission electron micrographs of intracellular single gold nanoparticles, they have the shape of a polyhedron.     

Gold-coated iron oxide nanoparticles as a T2 contrast agent in magnetic resonance imaging

Gold-coated iron oxide (Fe3O4) nanoparticles were synthesized for use as a T2 contrast agent in magnetic resonance imaging (MRI). The coated nanoparticles were spherical in shape with an average diameter of 20 nm. The gold shell was about 2 nm thick. The bonding status of the gold on the nanoparticle surfaces was checked using a Fourier transform infrared spectrometer (FTIR). The FTIR spectra confirmed the attachment of homocysteine, in the form of thiolates, to the Au shell of the Au-Fe3O4 nanoparticles. The relaxivity ratio, R2/R1, for the coated nanoparticles was 3-fold higher than that of a commercial contrast agent, Resovist, which showed the potential for their use as a T2 contrast agent with high efficacy. In animal experiments, the presence of the nanoparticles in rat liver resulted in a 71% decrease in signal intensity in T2-weighted MR images, indicating that our gold-coated iron oxide nanoparticles are suitable for use as a T2 contrast agent in MRI.     

Size-dependent cytotoxicity of gold nanoparticles

Gold nanoparticles are widely used in biomedical imaging and diagnostic tests. Based on their established use in the laboratory and the chemical stability of Au(0), gold nanoparticles were expected to be safe. The recent literature, however, contains conflicting data regarding the cytotoxicity of gold nanoparticles. Against this background a systematic study of water-soluble gold nanoparticles stabilized by triphenylphosphine derivatives ranging in size from 0.8 to 15 nm is made. The cytotoxicity of these particles in four cell lines representing major functional cell types with barrier and phagocyte function are tested. Connective tissue fibroblasts, epithelial cells, macrophages, and melanoma cells prove most sensitive to gold particles 1.4 nm in size, which results in IC(50) values ranging from 30 to 56 microM depending on the particular 1.4-nm Au compound-cell line combination. In contrast, gold particles 15 nm in size and Tauredon (gold thiomalate) are nontoxic at up to 60-fold and 100-fold higher concentrations, respectively. The cellular response is size dependent, in that 1.4-nm particles cause predominantly rapid cell death by necrosis within 12 h while closely related particles 1.2 nm in diameter effect predominantly programmed cell death by apoptosis.     

Surface enhanced absorption and transmission from dye coated gold nanoparticles in thin films

Absorption spectra of gold nanoisland thin film and the composite film of gold having thin coating of Methylene Blue and Rh6G dyes have been studied. Thin gold nanoisland film shows surface plasmon resonance (SPR) peak in the visible wavelength range, which shifts to near infrared with an increase in the thickness of the film. It was found that thin film of gold consists of nanoparticles of different size and shape, particularly nanorods of noncylindrical shapes. A linear relation was found between SPR peak wavelength and the aspect ratio of the nanoparticles in gold thin film. Effective medium refractive index of the gold film is estimated to be ~2.5, which decreases with an increase in film thickness. The coating of dyes on gold films splits the SPR peak with an enhanced absorption. Enhancement in absorption of composite film is maximal when the dye absorption peak coincides with the SPR peak; otherwise enhancement in transmission is observed for all the wavelength range. Absorption amplitude of composite film peaks increase with an increase in the gold film thickness, which tend toward saturation for film thickness of ≥6 nm. A correlation shows that absorption spectra can be described by the Maxwell Garnett theory, when the gold nanoparticles have a nearly spherical shape for very thin film (≤6 nm).     

Synthesis of gold nanoparticles using aqueous extract of Calotropis procera latex

This is an account of the use of aqueous extract of the latex of Calotropis procera for the synthesis of gold nanoparticles. UV-Vis spectroscopic studies of the products resulting from reaction between the aqueous latex extract and chlorauric acid indicated the successful synthesis of gold nanoparticles. Reaction parameters viz. concentration of latex extract and reaction time were optimized for maximum yield of gold nanoparticles. Effect of reaction temperature on the synthesis rate of the particles and their optical properties was also studied. Transmission electron microscopic (TEM) studies of the particles revealed the dominance of spherical particles. Mean particle size distribution was found to be 22 ± 10 nm. Crystalline nature of the particles was confirmed from X-ray diffractrograms. FT-IR analysis and protein coagulation test of the gold nanoparticles confirmed capping behaviour of the latex proteins that contributed to their long term stability (6 months) in aqueous medium. Toxicity of the particles was tested on three cell lines, HeLa, A549 and BHK21. The method exploits a cheap and easily available biomaterial not explored so far for the synthesis of metallic nanoparticles.     

Ascorbic acid-mediated synthesis and characterisation of iron oxide/gold core–shell nanoparticles.

The current article reports on providing surface modification of magnetic nanoparticles with gold to provide stability against aggregation. Gold-coated magnetite nanoparticles were synthesised to combine both magnetic as well as surface plasma resonance (SPR) properties in a single moiety. The nanocomposites were produced by reduction (using ascorbic acid) of gold chloride on to the surface of iron oxide nanoparticles. Ascorbic acid not only acts as a reducing agent, but also the oxidised form of ascorbic acid i.e. Dehydro-ascorbic acid acts as a capping agent to impart stability to as synthesised gold-coated iron oxide nanocomposites. The synthesised nanocomposite was monodispersed with a mean particle size of around 16 nm and polydispersity index of 0.190. X-ray diffraction analysis confirms presence of gold on the surface of magnetite nanoparticles. The synthesised nanocomposites had a total organic content of around 3.2% w/w and also showed a shifted SPR peak at 546 nm as compared to gold nanoparticles (528 nm). Both uncoated and gold-coated magnetite exhibited superparamagnetic behaviour at room temperature. Upon coating with gold shell, saturation magnetisation of iron oxide nanoparticles decreases from 42.806 to 3.54 emu/gram.     

Synthesis of metallic nanoparticles protected with N,N,N-trimethyl chitosan chloride via a relatively weak affinity

A water-soluble cationic chitosan derivative, N,N,N-trimethyl chitosan chloride (TMC), was synthesized and used as a stabilizing reagent for the synthesis of highly stable Au, Ag and Pt nanoparticles in a single-phase of neutral aqueous solution. The morphology and stability of metallic nanoparticles were evaluated by transmission electron microscopy and UV-vis spectroscopy. The results showed that well-dispersed metallic nanoparticles have a spherical morphology with diameters of about 3 ± 0.5?nm. The prepared gold nanoparticles are stable in the aqueous solution (no significant changes in their morphology and size within 10?months) due to repulsion between the charged polymer shell coatings around the metallic nanoparticles. The relatively low affinity of TMC on gold nanoparticles was confirmed by using a ligand exchange experiment. The mechanism stabilizing the chitosan derivative and the neighbouring gold nanoparticles was identified by FTIR, (1)H NMR and (13)C NMR measurements.     

Poly(amidoamine)-G5 dendrimers/noble metal gold hybrid nanoparticles prepared by γ-ray irradiation

In the absence of a chemical reductant and other protective reagents, noble metal gold hybrid nanoparticles are successfully prepared by ⁶⁰Co γ-ray irradiation using fifth-generation poly(amidoamine) dendrimer with surface amine-terminated group as polymeric template. The zerovalent gold is of spherical structure and the particle size is on nanometer scale range of 3-12 nm. The size distribution of gold nanoparticles displays multidispersity. The results of FTIR show that interactions between dendrimer template and gold nanoparticle exist in intra-molecule and inter-molecule of fifth-generation poly(amidoamine) dendrimer.     

Cytotoxicity of spherical gold nanoparticles synthesised using aqueous extracts of aerial roots of Rhaphidophora aurea (Linden ex Andre) intertwined over Lawsonia inermis and Areca catechu on MCF‐7 cell line

A facile synthesis of gold nanoparticles (GNPs) using the aqueous extracts of the aerial roots of Rhaphidophora aurea (Linden ex Andre) intertwined over Lawsonia inermis and Areca catechu was carried out under different conditions, namely room temperature, higher temperature, sonication, solar irradiation and pH variation. The surface plasmon resonance (SPR) band at 536 and 575 nm obtained in UV–visible spectrum revealed the formation of AuNP''s. The sharp SPR band of the synthesised nanogold indicates the formation of spherical‐shaped and uniform‐sized nanoparticles. The TEM analysis revealed spherical nanogold particles of size 35 and 10 nm for MM and MP extracts. The secondary metabolites present in the aqueous extract are suggested to be responsible for the reduction of metal ions to metal nanoparticles as evidenced from results of FTIR analysis. Rapid synthesis of GNPs by sunlight is the production of microscopic grains of gold due to the dissociation of gold chloride. This may induce the reaction between secondary metabolites and gold chloride solutions and results in GNPs. The cytotoxic activity of the synthesised nanogold studied against human breast cancer cells (MCF‐7) by 3‐[4,5‐dimethylthiazol‐2‐yl]2,5‐diphenyltetrazolium bromide assay showed significant activity at higher concentration.Inspec keywords: toxicology, gold, nanoparticles, nanomedicine, nanofabrication, cellular biophysics, particle size, surface plasmon resonance, ultraviolet spectra, visible spectra, transmission electron microscopy, dissociation, cancer, biological organs, Fourier transform infrared spectraOther keywords: cytotoxicity, spherical gold nanoparticles, aerial roots, Rhaphidophora aurea, Linden ex Andre, Lawsonia inermis, Areca catechu, MCF‐7 cell line, sonication, solar irradiation, pH variation, surface plasmon resonance, UV–visible spectrum, spherical‐shaped nanoparticles, uniform‐sized nanoparticles, TEM analysis, spherical nanogold particles, secondary metabolites, metal ions, FTIR analysis, microscopic grains, dissociation, gold chloride solutions, cytotoxic activity, human breast cancer cells, 3‐[4,5‐dimethylthiazol‐2‐ yl]2,5‐diphenyltetrazolium bromide assay, wavelength 536 nm, wavelength 575 nm, Au     

The role of surface charge on the uptake and biocompatibility of hydroxyapatite nanoparticles with osteoblast cells

The objective of this study is to evaluate the effect of hydroxyapatite (HAP) nanoparticles with different surface charges on the cellular uptake behavior and in vitro cell viability and proliferation of MC3T3-E1 cell lines (osteoblast). The nanoparticles' surface charge was varied by surface modification with two carboxylic acids: 12-aminododecanoic acid (positive) and dodecanedioic acid (negative). The untreated HAP nanoparticles and dodecanoic acid modified HAP nanoparticles (neutral) were used as the control. X-ray diffraction (XRD) revealed that surface modifications by the three carboxylic acids did not change the crystal structure of HAP nanoparticles; Fourier transform infrared spectroscopy (FT-IR) confirmed the adsorption and binding of the carboxylic acids on the HAP nanoparticles' surfaces; and zeta potential measurement confirmed that the chemicals successfully modified the surface charge of HAP nanoparticles in water based solution. Transmission electron microscopy (TEM) images showed that positively charged, negatively charged and untreated HAP nanoparticles, with similar size and shape, all penetrated into the cells and cells had more uptake of HAP nanoparticles with positive charge compared to those with negative charge, which might be attributed to the attractive or repulsive interaction between the negatively charged cell membrane and positively/negatively charged HAP nanoparticles. The neutral HAP nanoparticles could not penetrate the cell membrane due to their larger size. MTT assay and LDH assay results indicated that as compared with the polystyrene control, greater cell viability and cell proliferation were measured on MC3T3-E1 cells treated with the three kinds of HAP nanoparticles (neutral, positive, and untreated), among which positively charged HAP nanoparticles showed the strongest improvement for cell viability and cell proliferation. In summary, the surface charge of HAP nanoparticles can be modified to influence the cellular uptake of HAP nanoparticles and the different uptake also influences the behavior of cells. These in vitro results may also provide useful information for investigations of HAP nanoparticle applications in gene delivery and intracellular drug delivery.     

Cetyltrimethylammonium bromide-modified spherical and cube-like gold nanoparticles as extrinsic Raman labels in surface-enhanced Raman spectroscopy based heterogeneous immunoassays

This paper reports on the characterization and preliminary comparison of gold nanoparticles of differing surface modification and shape when used as extrinsic Raman labels (ERLs) in high-sensitivity heterogeneous immunoassays based on surface enhanced Raman scattering (SERS). ERLs are gold nanoparticles coated with an adlayer of an intrinsically strong Raman scatterer, followed by a coating of a molecular recognition element (e.g., antibody). Three types of ERLs, all with a nominal size of approximately 30 nm, were fabricated by using spherical citrate-capped gold nanoparticles (sp-cit-Au NPs), spherical CTAB-capped gold nanoparticles (sp-CTAB-Au NPs), or cube-like CTAB-capped gold nanoparticles (cu-CTAB-Au NPs) as cores. The performance of these particles was assessed via a sandwich immunoassay for human IgG in phosphate buffered saline. The ERLs fabricated with sp-CTAB-Au NPs as cores proved to be more than 50 times more sensitive than those with sp-cit-Au NPs as cores; the same comparison showed that the ERLs with cu-CTAB-Au NPs as cores were close to 200 times more sensitive. Coupled with small differences in levels of nonspecific adsorption, these sensitivities translated to a limit of detection (LOD) of 94, 2.3, and 0.28 ng/mL, respectively, for the detection of human IgG in the case of sp-cit-Au NPs, sp-CTAB-Au NPs, and cu-CTAB-Au NPs. The LOD of the cu-CTAB-Au NPs is therefore approximately 340 times below that for the sp-cit-Au NPs. Potential applications of these labels to bioassays are briefly discussed.     

Plasmonic intravascular photoacoustic imaging for detection of macrophages in atherosclerotic plaques

To detect macrophages in atherosclerotic plaques, plasmonic gold nanoparticles are introduced as a contrast agent for intravascular photoacoustic imaging. The phantom and ex vivo tissue studies show that the individual spherical nanoparticles, resonant at 530 nm wavelength, produce a weak photoacoustic signal at 680 nm wavelength while photoacoustic signal from nanoparticles internalized by macrophages is very strong due to the plasmon resonance coupling effect. These results suggest that intravascular photoacoustic imaging can assess the macrophage-mediated aggregation of nanoparticles and therefore identify the presence and the location of nanoparticles associated with macrophage-rich atherosclerotic plaques.     

苯乙烯-马来酸酐共聚物/金纳米微粒的合成

以氯金酸为原料,DMF(N,N-二甲基甲酰胺)为溶剂及还原剂,苯乙烯-马来酸酐共聚物为大分子稳定剂,合成了金纳米微粒。通过紫外-可见吸收光谱、透射电子显微镜等方法对纳米金样品进行了表征。结果表明:所得到的金纳米微粒可以在520 nm~530 nm范围内产生明显的纳米金所具有的特征等离子共振吸收峰,金纳米微粒的尺寸在3 nm~5nm且具有较窄的分布,证明苯乙烯-马来酸酐共聚物可以对金纳米微粒表面产生较好的修饰作用,从而为制备纳米金材料提供了一种新的途径。     

Biosynthesis of Au nanoparticles using cumin seed powder extract

Cumin seed was investigated for synthesis of gold nanoparticles. Polydispersed particles were obtained at pH 3 and 30 degrees C, and the effect of temperature and pH on synthesis of gold nanoparticles was analyzed. TEM images showed that amount of platelets formed were predominant at lower temperature where as more number of monodispersed spherical particle of size 1-10 nm were perceived at high temperatures. The gold nanoparticles particles formed at higher pH were stable, uniform and spherical in shape. XPS analysis showed the presence of pure gold nanoparticles.     

Paramagnetic silica-coated gold nanoparticles

Water soluble gold nanoparticles, obtained by the reduction of the gold (III) chloride with sodium borohydride in the presence of citric acid or thioctic acid, were covered with a paramagnetic silica layer using the Stober method, yielding a hybrid metallic-inorganic nanomaterial (gold nanoparticles, with an average size of 5 nm, embedded into silica nanoparticles, with an average size of 100 nm). The paramagnetic silica layer was formed by copolymerization of a paramagnetic silica precursor (derived from 3-aminopropyltrimethoxysilane) with tetramethyorthosilicate. The paramagnetic silica precursor was obtained by coupling 3-aminopropyltrimethoxysilane with 3-carboxy-proxyl free radical. TEM pictures show that each silica nanoparticle of about 100 nm in size embedded about 10 gold nanoparticles. These hybrid nanoparticles are quite stable and exhibit the expected paramagnetic characteristics, as seen by electron paramagnetic resonance. The accessibility of methanol through the silica layer was also studied. Depending on the capping ligands of the gold nanoparticles (citric or thioctic acid), different silica networks are formed, as seen by the mobility of the spin-label inside the silica layer. The EPR spectra showed that the paramagnetic silica layer is very robust and the mobility of the spin-probe inside the silica layer is very little affected by methanol. However, if spin-labeled thioctic acid protected gold nanoparticles were used in the material synthesis, the mobility of the spins attached to the gold surface is quite high in the presence of methanol, while the spins embedded into the silica layer remains immobilized.     

Soybeans as a phytochemical reservoir for the production and stabilization of biocompatible gold nanoparticles

The present study demonstrates an unprecedented green process for the production of gold nanoparticles by simple treatment of gold salts with soybean extracts. Reduction capabilities of antioxidant phytochemicals present in soybean and their ability to reduce gold salts chemically to nanoparticles with subsequent coating of proteins and a host of other phytochemicals present in soybean on the freshly generated gold nanoparticles are discussed. The new genre of green nanoparticles exhibit remarkable in vitro stability in various buffers including saline, histidine, HSA, and cysteine solutions. MTT assays reveal that the green gold nanoparticles are nontoxic and thus provide excellent opportunities for their applications in nanomedicine for molecular imaging and therapy. The overall strategy described herein for the generation of gold nanoparticles meets all 12 principles of green chemistry, as no "man-made" chemicals, other than the gold salts, are used in the green nanotechnological process.     

Mechanism for the Cellular Uptake of Targeted Gold Nanorods of Defined Aspect Ratios

Biomedical applications of non‐spherical nanoparticles such as photothermal therapy and molecular imaging require their efficient intracellular delivery, yet reported details on their interactions with the cell remain inconsistent. Here, the effects of nanoparticle geometry and receptor targeting on the cellular uptake and intracellular trafficking are systematically explored by using C166 (mouse endothelial) cells and gold nanoparticles of four different aspect ratios (ARs) from 1 to 7. When coated with poly(ethylene glycol) strands, the cellular uptake of untargeted nanoparticles monotonically decreases with AR. Next, gold nanoparticles are functionalized with DNA oligonucleotides to target Class A scavenger receptors expressed by C166 cells. Intriguingly, cellular uptake is maximized at a particular AR: shorter nanorods (AR = 2) enter C166 cells more than nanospheres (AR = 1) and longer nanorods (AR = 4 or 7). Strikingly, long targeted nanorods align to the cell membrane in a near‐parallel manner followed by rotating by ≈90° to enter the cell via a caveolae‐mediated pathway. Upon cellular entry, targeted nanorods of all ARs predominantly traffic to the late endosome without progressing to the lysosome. The studies yield important materials design rules for drug delivery carriers based on targeted, anisotropic nanoparticles.     

Synthesis, characterization, and direct intracellular imaging of ultrasmall and uniform glutathione-coated gold nanoparticles

Gold nanoparticles (AuNPs) with core sizes below 2 nm and compact ligand shells constitute versatile platforms for the development of novel reagents in nanomedicine. Due to their ultrasmall size, these AuNPs are especially attractive in applications requiring delivery to crowded intracellular spaces in the cytosol and nucleus. For eventual use in vivo, ultrasmall AuNPs should ideally be monodisperse, since small variations in size may affect how they interact with cells and how they behave in the body. Here we report the synthesis of ultrasmall, uniform 144-atom AuNPs protected by p-mercaptobenzoic acid followed by ligand exchange with glutathione (GSH). Quantitative scanning transmission electron microscopy (STEM) reveals that the resulting GSH-coated nanoparticles (Au(GSH)) have a uniform mass distribution with cores that contain 134 gold atoms on average. Particle size dispersity is analyzed by analytical ultracentrifugation, giving a narrow distribution of apparent hydrodynamic diameter of 4.0 ± 0.6 nm. To evaluate the nanoparticles' intracellular fate, the cell-penetrating peptide TAT is attached noncovalently to Au(GSH), which is confirmed by fluorescence quenching and isothermal titration calorimetry. HeLa cells are then incubated with both Au(GSH) and the Au(GSH)-TAT complex, and imaged without silver enhancement of the AuNPs in unstained thin sections by STEM. This imaging approach enables unbiased detection and quantification of individual ultrasmall nanoparticles and aggregates in the cytoplasm and nucleus of the cells.     

Layer‐by‐Layer Coated Gold Nanoparticles: Size‐Dependent Delivery of DNA into Cells

Because nanoparticles are finding uses in myriad biomedical applications, including the delivery of nucleic acids, a detailed knowledge of their interaction with the biological system is of utmost importance. Here the size‐dependent uptake of gold nanoparticles (AuNPs) (20, 30, 50 and 80 nm), coated with a layer‐by‐layer approach with nucleic acid and poly(ethylene imine) (PEI), into a variety of mammalian cell lines is studied. In contrast to other studies, the optimal particle diameter for cellular uptake is determined but also the number of therapeutic cargo molecules per cell. It is found that 20 nm AuNPs, with diameters of about 32 nm after the coating process and about 88 nm including the protein corona after incubation in cell culture medium, yield the highest number of nanoparticles and therapeutic DNA molecules per cell. Interestingly, PEI, which is known for its toxicity, can be applied at significantly higher concentrations than its IC₅₀ value, most likely because it is tightly bound to the AuNP surface and/or covered by a protein corona. These results are important for the future design of nanomaterials for the delivery of nucleic acids in two ways. They demonstrate that changes in the nanoparticle size can lead to significant differences in the number of therapeutic molecules delivered per cell, and they reveal that the toxicity of polyelectrolytes can be modulated by an appropriate binding to the nanoparticle surface.     

Hollow spherical nucleic acids for intracellular gene regulation based upon biocompatible silica shells

Cellular transfection of nucleic acids is necessary for regulating gene expression through antisense or RNAi pathways. The development of spherical nucleic acids (SNAs, originally gold nanoparticles functionalized with synthetic oligonucleotides) has resulted in a powerful set of constructs that are able to efficiently transfect cells and regulate gene expression without the use of auxiliary cationic cocarriers. The gold core in such structures is primarily used as a template to arrange the nucleic acids into a densely packed and highly oriented form. In this work, we have developed methodology for coating the gold particle with a shell of silica, modifying the silica with a layer of oligonucleotides, and subsequently oxidatively dissolving the gold core with I(2). The resulting hollow silica-based SNAs exhibit cooperative binding behavior with respect to complementary oligonucleotides and cellular uptake properties comparable to their gold-core SNA counterparts. Importantly, they exhibit no cytotoxicity and have been used to effectively silence the eGFP gene in mouse endothelial cells through an antisense approach.