A colorimetric chemosensor for sensitive and selective detection of copper(II) ions based on catalytic oxidation of 1-naphthylamine.

Rapid on-site detection of copper(II) ions (Cu2+) with high sensitivity and selectivity is of great significance in the safety monitoring of drinking water and food. Colorimetric detection is a robust fast determination method with the main drawback of low sensitivity. Herein, we developed a colorimetric chemosensor based on a colored polymer product. Via a Cu-Fenton mechanism, 1-naphthylamine (α-NA) was oxidized by H2O2 and brownish-red poly(1-naphthylamine) (PNA) was produced. The obtained Cu2+ sensor showed a linear response from 0.05 µM to 7 µM, with a detection limit of 62 nM. Our findings expanded chromogenic reaction types for colorimetric detection.

Modulation of plasmonic chiral shell growth on gold nanorods via nonchiral surfactants.

Recently, in combination with seed-mediated growth, thiolated chiral molecule-guided growth has shown great promise in obtaining chiral plasmonic nanostructures. Previously, with the assistance of chiral cysteines (Cys), we realized helical growth of plasmonic shells on gold nanorod (AuNR) seeds dispersed in cetyltrimethylammonium bromide (CTAB) solution. Herein, we further studied the roles of non-chiral cationic surfactants in tuning the helical growth. Both the counter anion and the hydrocarbon chain length of the surfactants were found to affect the formation of helical shells greatly. In particular, we exhibited surfactant-modulated conversion of the chiral shell deposition mode between layer growth and island growth. By optimizing growth conditions, an obvious plasmonic circular dichroism (PCD) response could be achieved for the island helical shell. Our findings demonstrated promising potential of nanochemical synthesis in fabricating chiral plasmonic nanostructures with small structural sizes.

Bottom-Up Synthesis of Helical Plasmonic Nanorods and Their Application in Generating Circularly Polarized Luminescence.

Chiral growth and chirality transfer associated with plasmonic nanostructures have rejuvenated the field of chirality. As the precise regioselective growth of inorganic crystals into chiral shapes at the nanoscale is extremely challenging, "bottom-up" synthesis of intrinsically chiral nanoparticles with structural stability is obviously attractive and important. With the thiolated bimolecular cosurfactants, we demonstrated a chemical strategy for the synthesis of intrinsically helical plasmonic nanorods (HPNRs) with strong and tailorable plasmonic circular dichroism (PCD) responses, deriving from the zwitterionic interactions between the -NH3+ and -COO- groups of the cysteine molecules (Cys). The influence of structural parameters of HPNRs on PCD responses was analyzed systematically by theoretical simulations. Among the different structural parameters, the pitch depth was found to have the greatest impact on the PCD signals, in agreement with the experimental results. Moreover, the obtained HPNRs with the strong, tunable, and stable chiroptical properties were found to be able to induce circularly polarized luminescence of achiral luminophores. Due to the generality of this effect, this chiral plasmonic nanostructure may have great potential for use in the fields of chiral sensors, chiral catalysis, and displays.

Improving peroxidase activity of gold nanorod nanozymes by dragging substrates to the catalysis sites via cysteine modification.

Surface chemistry control is a key means to improve substrate selectivity and enhance catalytic activity of nanozymes, a kind of novel artificial enzymes. Herein, we demonstrated that apart from chemical properties of functional groups, their spatial distance to the catalytic sites is also very important to improve the catalytic performance of nanozymes. Using cetyltrimethylammonium bromide (CTAB) coated gold nanorods (AuNR) as the example, we showed that cysteine (Cys) surface modification can greatly enhance the peroxidase activity of AuNR for the oxidation of substrate 3,3',5,5'-tetramethylbenzidine (TMB). By using cysteine derivatives, the key role of the carboxylic group in cysteine is revealed in improving substrate binding and activity enhancement. The electrostatic interactions of carboxylic groups from adsorbed cysteine molecules with protonated amino groups of TMB bring TMB molecules to the surface Au active sites and thus markedly increase catalytic activity. In contrast, despite having two carboxylic groups, glutathione (GSH) surface modification only leads to quite limited improvement of catalytic activity. We speculated that due to large molecular size of GSH, the spatial distance between TMB-GSH and Au is larger than that between TMB-Cys and Au. Furthermore, Raman characterization indicated that at high Cys coverage, they form patches on rod surface via zwitterionic interactions, which may give additional benefits by decreasing the steric hindrance of TMB diffusion to surface Au atom sites. In all, our study highlights the importance of fine surface tuning in the design of nanozymes.

Constructing chiral gold nanorod oligomers using a spatially separated sergeants-and-soldiers effect.

A sergeants-and-soldiers (S&S) effect is very useful to the fabrication of supramolecular chirality. This strategy has not yet been explored in the construction of chiral plasmonic superstructures. Herein, we demonstrate a spatially separated S&S effect in fabricating plasmonic superstructures and modulating their chiroptical responses. Specifically, chiral cysteine (Cys) molecules, acting as sergeants, are sandwiched between a gold nanorod (AuNR) core and a Au shell via AuNR-templated Au overgrowth. Cationic surfactants, CTAB (cetyltrimethylammonium bromide) or CPC (cetylpyridinium chloride), are modified on the AuNR@Cys@Au shell surface, thus spatially separating from the chiral sergeants. During the assembly process, the surfactants act as soldiers which could transfer and amplify the local chirality induced by the adsorbed chiral molecules from the plasmonic monomers to the oligomers. Huge PCD signals could be achieved in the plasmonic oligomers by finely tuning chiral sergeants and achiral soldiers, indicating the feasibility of the S&S effect in fabricating chiral plasmonic superstructures.

Initiation of protective autophagy in hepatocytes by gold nanorod core/silver shell nanostructures.

The high reactivity of silver nanoparticles leads to their broad applications in the anti-bacterial field; however, the safety of silver nanoparticles has attracted increasing public attention. After exposure to silver nanoparticles in vivo, the liver serves as their potential deposition site; however the potential biological effects of such nanoparticles on hepatocytes at low dosages are not well understood. Here, we study the interaction between gold nanorod core/silver shell nanostructures (Au@Ag NRs) and human hepatocytes, HepG2 cells, and determine that Au@Ag NRs at sub-lethal doses can induce autophagy. After uptake, Au@Ag NRs mainly localize in the lysosomes where they release silver ions and promote the production of reactive oxygen species (ROS). The ROS then suppress the AKT-mTOR signaling pathway and activate autophagy. In addition, oxidative stress results in lysosomal impairment, causing decreased ability for lysosomal digestion. Moreover, oxidative stress also affects the structure and function of mitochondria, leading to the initiation of protective autophagy to eliminate the damaged mitochondrion. Our study shows that at sub-lethal dosages, silver nanomaterials may alter the physiological functions of hepatic cells by activating protective autophagy and cause potential health risks, indicating that cautious consideration of the safety of nanomaterials for certain applications is necessary.

Corona of Thorns: The Surface Chemistry-Mediated Protein Corona Perturbs the Recognition and Immune Response of Macrophages.

The significance of protein coronas on the biological fates of nanoparticles has been widely recognized. Therefore, the alterations on biological effects caused by protein coronas need systemic study and interpretation to design novel safe and efficient nanomedicines. In the present study, we present a comprehensive quantitative analysis of the protein coronas on gold nanorods  modified with various surface ligands of different chemical compositions and charges. The design of surface ligands is of utmost importance for the functionalization of nanoparticles, and further, the ligand-induced biological identity determines the fate of nanoparticles in the human body. We found that the surface chemistry influences the composition of the protein corona more profoundly than surface charge. Since the first and most important challenge for administrated nanomedicines is navigating the interaction with macrophages, we further investigated how the surface chemistry-induced specific protein corona affects the phagocytosis and immune responses of macrophages exposed to the corona-nanoparticle complexes. Our results reveal that the protein corona alters the internalization pathways of gold nanorods by macrophages via the interactions of the predominant coronal proteins with specific receptors on the cell membrane. The cytokine secretion profile of macrophages is also highly dependent on the adsorption pattern of the protein corona. The more abundant proteins involved in immune responses, such as acute phase, complement, and tissue leakage proteins, present in the acquired nanoparticle corona, the more macrophage interleukin-1ß (IL-1ß) released is stimulated. The ligand-protein corona composition-immune response coefficient analysis may serve next-generation nanomedicines with high efficiency and good safety for better clinical translation.

Cytotoxicity and Cellular Responses of Gold Nanorods to Smooth Muscle Cells Dependent on Surface Chemistry Coupled Action.

Gold nanorods (AuNRs), with their unique physicochemical properties, are recognized as promising materials for biomedical applications. Chemical modification of their surfaces is attracting increasing attention with regard to cytotoxicity and cellular uptake. Herein, the toxicological effects of three types of polymer-coated AuNRs, which are cetyltrimethylammonium bromide-coated AuNRs, polystyrene sulphonate-coated AuNRs, and poly(diallyldimethyl ammonium chloride-coated AuNRs (PDDAC-AuNRs), on vascular smooth muscle cells (VSMCs) are investigated. The results show significantly different effects on VSMCs with different surface coatings. PDDAC-AuNRs, which were nontoxic in cancer cells in previous reports, display extreme toxicity to VSMCs. Initial contact between AuNRs and cell membranes is the important step in AuNRs cellular uptake. Force spectroscopy based on atomic force microscopy is exploited to study interactions between AuNRs and VSMCs membrane in the absence or presence of a corona on the AuNRs surface. The results show that the binding force and binding probability between AuNRs and membranes are closely related to cytotoxicity and cellular responses. These findings highlight the importance of assessing nanoparticle cytotoxicity in somatic cells for medical applications.

Fabricating chiroptical starfruit-like Au nanoparticles via interface modulation of chiral thiols.

The surface/interface matters as the size of materials enters the nanoscale. Control of surface/interface, therefore, plays an important role in creating novel nanostructures with unusual properties and in obtaining devices with high performance. Herein, we demonstrate unique interface regulation in fabricating nanostructures with strong plasmonic circular dichroism (PCD). With chiral cysteine (Cys) as surface-modulating molecules, starfruit-like Au nanoparticles (NPs) with high PCD responses are obtained via Au overgrowth on Au nanorods (AuNRs). Pre-incubation of the AuNRs with Cys is vital in achieving strong and reproducible PCD responses. Instead of contributing to PCD signals, the pre-adsorbed Cys molecules are found to play a major role in manipulating the Au growth mode and thus the formation of hotspots within the shell. Strong PCD signal mainly comes from the entrapped Cys molecules within the hotspots and is enhanced via local field effect. The distinct roles of the same ligands at different surfaces/interfaces are elucidated. Furthermore, our findings contribute to the strategy of utilizing interface modulation to fabricate complex nanostructures with novel properties.

Interference of Steroidogenesis by Gold Nanorod Core/Silver Shell Nanostructures: Implications for Reproductive Toxicity of Silver Nanomaterials.

As a widely used nanomaterial in daily life, silver nanomaterials may cause great concern to female reproductive system as they are found to penetrate the blood-placental barrier and gain access to the ovary. However, it is largely unknown about how silver nanomaterials influence ovarian physiology and functions such as hormone production. This study performs in vitro toxicology study of silver nanomaterials, focusing especially on cytotoxicity and steroidogenesis and explores their underlying mechanisms. This study exposes primary rat granulosa cells to gold nanorod core/silver shell nanostructures (Au@Ag NRs), and compares outcomes with cells exposed to gold nanorods. The Au@Ag NRs generate more reactive oxygen species and reduce mitochondrial membrane potential and less production of adenosine triphosphate. Au@Ag NRs promote steroidogenesis, including progesterone and estradiol, in a time- and dose-dependent manner. Chemical reactivity and transformation of Au@Ag NRs are then studied by electron spin resonance spectroscopy and X-ray absorption near edge structure, which analyze the generation of free radical and intracellular silver species. Results suggest that both particle-specific activity and intracellular silver ion release of Au@Ag NR contribute to the toxic response of granulosa cells.

Gd-Hybridized Plasmonic Au-Nanocomposites Enhanced Tumor-Interior Drug Permeability in Multimodal Imaging-Guided Therapy.

An original gadolinium-hybridized plasmonic gold nanocomposite is fabricated to provide an insightful and attractive strategy to overcome both the physiological and pathological barriers of tumor, and increase the transportation and permeability of imaging agents and drugs in tumor interior for achieving high-sensitive multimodal imaging and simultaneously improving the therapeutic efficacy of cancer.

ZnO Nanoparticles Treatment Induces Apoptosis by Increasing Intracellular ROS Levels in LTEP-a-2 Cells.

Owing to the wide use of novel nanoparticles (NPs) such as zinc oxide (ZnO) in all aspects of life, toxicological research on ZnO NPs is receiving increasing attention in these days. In this study, the toxicity of ZnO NPs in a human pulmonary adenocarcinoma cell line LTEP-a-2 was tested in vitro. Log-phase cells were exposed to different levels of ZnO NPs for hours, followed by colorimetric cell viability assay using tetrazolium salt and cell survival rate assay using trypan blue dye. Cell morphological changes were observed by Giemsa staining and light microscopy. Apoptosis was detected by using fluorescence microscopy and caspase-3 activity assay. Both intracellular reactive oxygen species (ROS) and reduced glutathione (GSH) were examined by a microplate-reader method. Results showed that ZnO NPs (≥ 0.01 µg/mL) significantly inhibited proliferation (P < 0.05) and induced substantial apoptosis in LTEP-a-2 cells after 4 h of exposure. The intracellular ROS level rose up to 30-40% corresponding to significant depletion (approximately 70-80%) in GSH content in LTEP-a-2 cells (P < 0.05), suggesting that ZnO NPs induced apoptosis mainly through increased ROS production. This study elucidates the toxicological mechanism of ZnO NPs in human pulmonary adenocarcinoma cells and provides reference data for application of nanomaterials in the environment.

Plasmonic polymers with strong chiroptical response for sensing molecular chirality.

We report on the chiroptical transfer and amplification effect observed in plasmonic polymers consisting of achiral gold nanorod monomers linked by cysteine chiral molecules in an end-to-end fashion. A new strategy for controlling the hot spots based circular dichroism (CD)-active sites in plasmonic polymers was developed to realize tailored and reproducible chiroptical activity in a controlled way. We showed that by regulating the bond angles between adjacent nanorods and the degree of polymerization in the linear plasmonic polymer, weak molecular chirality in the ultraviolet spectral region can be amplified by more than two orders of magnitude via the induced CD response in the visible/near infrared region. We demonstrate that this plasmonic polymer can be used to provide not only the Raman "fingerprint" information for identifying the molecular identity but also the CD signatures for (i) resolving the enantiomeric pairs of cysteine molecules at a small quantity level, and (ii) quantifying the enantiomeric purity of the chiral analytes. Chiral analyses by chiroptically responsive plasmonic polymers may find important applications in bioscience and biomedicine.

Plasmonic circular dichroism in side-by-side oligomers of gold nanorods: the influence of chiral molecule location and interparticle distance.

Chiral metal nanostructures, which exhibit plasmonic circular dichroism (PCD), have great potential for the development of chiral sensors and devices. Previously, we developed a method for fabricating chiral gold-nanorod oligomers: the nanorods are linked by achiral molecules, while chiral molecules (e.g.l- or d-cysteine) on the Au surface exert a directional twisting force on the oligomers and thereby generate a PCD signal. In this paper, we investigate how the location of chiral molecules and the interparticle distance affect the PCD of the oligomers. Cysteine at the ends of the nanorods and those on the side were found to induce PCD with opposite signs. When we increased the interparticle distance, the PCD signal was weakened; in particular, cysteine at the ends lost the twisting effect. Besides introducing the twisting force at the Au surface, chiral molecules in the hydrophobic surfactant bilayer and those adsorbed outside the surfactants can also twist the oligomers and generate PCD signals. These findings not only provide guidelines to the manipulation of PCD signals, but also serve as a more elaborate platform for studying the nanoscale interactions between nanoparticles.

Identification and mRNA expression of two 17ß-hydroxysteroid dehydrogenase genes in the marine mussel Mytilus galloprovincialis following exposure to endocrine disrupting chemicals.

17ß-Hydroxysteroid dehydrogenases (17ß-HSDs) are multifunctional enzymes involved in the metabolism of steroids, fatty acids, retinoids and bile acid. In this study, two novel types of 17ß-HSDs (named as MgHsd17b10 and MgHsd17b12) were cloned from Mytilus galloprovincialis by using rapid amplification of cDNA ends (RACE) approaches. Sequence analysis showed that MgHsd17b10 and MgHsd17b12 encoded a polypeptide of 259 and 325 amino acids, respectively. Phylogenetic analysis revealed that MgHsd17b10 and MgHsd17b12 were evolutionarily clustered with other invertebrate 17ß-HSD type 10 and 17ß-HSD type 12 homologues. The MgHsd17b10 and MgHsd17b12 transcripts could be detected in all examined tissues with higher expression levels in digestive glands and gonad. After exposed to endocrine disrupting chemicals (Bisphenol A or 2,2',4,4'-tetrabromodiphenyl ether), the expression of MgHsd17b10 and MgHsd17b12 transcripts was both down-regulated in digestive glands. These findings suggest that MgHsd17b10 and MgHsd17b12 perhaps play an important role in the endocrine regulation of M. galloprovincialis.

Near infrared laser-induced targeted cancer therapy using thermoresponsive polymer encapsulated gold nanorods.

External stimuli, such as ultrasound, magnetic field, and light, can be applied to activate in vivo tumor targeting. Herein, we fabricated polymer encapsulated gold nanorods to couple the photothermal properties of gold nanorods and the thermo- and pH-responsive properties of polymers in a single nanocomposite. The activation mechamism was thus transformed from heat to near-infrared (NIR) laser, which can be more easily controlled. Doxorubicin, a clinical anticancer drug, can be loaded into the nanocomposite through electrostatic interactions with high loading content up to 24%. The nanocomposite's accumulation in tumor post systematic administration can be significantly enhanced by NIR laser irradiation, providing a prerequisite for their therapeutic application which almost completely inhibited tumor growth and lung metastasis. Since laser can be manipulated very precisely and flexibly, the nanocomposite provides an ideally versatile platform to simultaneously deliver heat and anticancer drugs in a laser-activation mechanism with facile control of the area, time, and dosage. The NIR laser-induced targeted cancer thermo-chemotherapy without using targeting ligands represents a novel targeted anticancer strategy with facile control and practical efficacy.

Revealing the binding structure of the protein corona on gold nanorods using synchrotron radiation-based techniques: understanding the reduced damage in cell membranes.

Regarding the importance of the biological effects of nanomaterials, there is still limited knowledge about the binding structure and stability of the protein corona on nanomaterials and the subsequent impacts. Here we designed a hard serum albumin protein corona (BSA) on CTAB-coated gold nanorods (AuNRs) and captured the structure of protein adsorption using synchrotron radiation X-ray absorption spectroscopy, microbeam X-ray fluorescent spectroscopy, and circular dichroism in combination with molecular dynamics simulations. The protein adsorption is attributed to at least 12 Au-S bonds and the stable corona reduced the cytotoxicity of CTAB/AuNRs. These combined strategies using physical, chemical, and biological approaches will improve our understanding of the protective effects of protein coronas against the toxicity of nanomaterials. These findings have shed light on a new strategy for studying interactions between proteins and nanomaterials, and this information will help further guide the rational design of nanomaterials for safe and effective biomedical applications.

Trafficking of gold nanorods in breast cancer cells: uptake, lysosome maturation, and elimination.

Gold nanorods (AuNRs) have been largely investigated driven by their promising potentials in drug delivery, imaging, and photodynamic therapy because of their distinctive physicochemical properties. It is widely known that AuNRs can be taken up by different cells, however, the trafficking of the nanorods in cells are less known. In this work, the behaviors and fate of AuNRs in the human breast cancer cell line MDA-MB-231 were intensively probed by transmission electron microscopy (TEM) with detailed time resolution, together with induced couple plasmon mass spectroscopy (ICP-MS), confocal microscopy, Western blot, and cell viability assay. We reveal that AuNRs enter the classic lysosome maturation through endocytosis and are sequestered in the vesicular system even during cell division. AuNRs can escape from the lysosomes occasionally and the escaped AuNRs are recycled back into the lysosomal system through cytoprotective autophagy. The dilution of AuNRs in cells is mainly attributed to the cell division rather than exocytosis, because expelled AuNRs can be re-endocytosed by the cells. The feature of vesicular restriction guarantees other organelles such as mitochondria and nucleus are exempted from the direct exposure to AuNRs.

Surface chemistry of gold nanorods: origin of cell membrane damage and cytotoxicity.

We investigated how surface chemistry influences the interaction between gold nanorods (AuNRs) and cell membranes and the subsequent cytotoxicity arising from them in a serum-free cell culture system. Our results showed that the AuNRs coated with cetyl trimethylammonium bromide (CTAB) molecules can generate defects in the cell membrane and induce cell death, mainly due to the unique bilayer structure of CTAB molecules on the surface of the rods rather than their charge. Compared to CTAB-capped nanorods, positively charged polyelectrolyte-coated, i.e. poly(diallyldimethyl ammonium chloride) (PDDAC), AuNRs show improved biocompatibility towards cells. Thus, the present results indicate that the nature of surface molecules, especially their packing structures on the surface of AuNRs rather than surface charge, play a more crucial role in determining cytotoxicity. These findings about interfacial interactions could also explain the effects of internalized AuNRs on the structures or functions of organelles. This study will help understanding of the toxic nature of AuNRs and guide rational design of the surface chemistry of AuNRs for good biocompatibility in pharmaceutical therapy.