Design of Gold Nanoparticles in Dendritic Cell‐Based Vaccines

The design of effective cancer vaccines must be able to activate dendritic cells (DCs) of the innate immune system in order to induce immunity to pathogens and cancer. DCs patrol the body and once they encounter antigens, they orchestrate a complex mechanism of events and signals that can alert the adaptive immune system to action. However, DC‐based vaccines remain a challenge in part because the source and quality of antigens, the DC targeting molecule, type of adjuvant, and delivery vehicle must be optimized to induce a robust immune response. Gold nanoparticles (AuNPs) have now entered clinical trials as carriers due to their ease of functionalization with antigens, adjuvants, and targeting molecules. This progress report discusses how AuNPs can influence DC activation and maturation, as well as their potential impact on T helper (Th) differentiation. Ultimately, successful AuNP‐based DC vaccines are able to induce phagocytosis, activation/maturation, migration, T cell costimulation, and cytokine secretion, which is named AuNP‐induced DC tuning (AuNP‐DC tuning). Although at its infancy, understanding the processes of AuNP‐DC tuning will give a better understanding of how best to engineer AuNPs and will redefine the next generation of DC‐based vaccines.     

金纳米粒子的放射增敏效应研究

系统阐述了与金纳米粒子(GNPs) 放射增敏效应相关实验的方法与结果、影响GNPs 放射增敏的因素、GNPs 放射增敏的细胞和动物实验表现及其相关机制。同时结合相关实验,分析和比较了15 nm 柠檬酸钠包被的GNPs 的放射增敏效应,发现GNPs 在高LET 的碳离子束和低LET 的X射线辐照下对Hela细胞的杀伤力随其浓度的增加而增大;在50% 的细胞存活率下,当GNPs 的质量浓度为7.5 g/mL时,其X射线的剂量减少率和碳离子的相对生物学效应值(RBE) 的增加率达到了最大,分别为65.3% 和43.6%,同时GNPs 共培养细胞24 和48 h 后,未出现细胞周期同步化的现象。This paper describes the methods and results of the previous experiments, the experimental phenomena of the cell and animal tests and the relative mechanisms on the radiosensitizing effect of GNPs. Together with our experiments, the radiosensitizing effects of 15 nm citrate-capped GNPs and related mechanisms are analyzed and compared, finding that Hela cell killing of GNPs increase along with their concentration after exposure to high- and low-LET radiation such as carbon ions and X-rays. In addition, the percentages of dose reduction of the X-rays and RBE increment of the carbon ions reached their maximums 65.3% and 43.6%, respectively,at 50% survival level when Hela cells were pre-treated with 7.5 g/mL GNPs. Moreover, Hela cells showed no cell-cycle synchronization after 24 and 48 h exposure to GNPs.     

Peptide‐Targeted Gold Nanoparticles for Photodynamic Therapy of Brain Cancer

Targeted drug delivery using epidermal growth factor peptide‐targeted gold nanoparticles (EGF_pep‐Au NPs) is investigated as a novel approach for delivery of photodynamic therapy (PDT) agents, specifically Pc 4, to cancer. In vitro studies of PDT show that EGF_pep‐Au NP‐Pc 4 is twofold better at killing tumor cells than free Pc 4 after increasing localization in early endosomes. In vivo studies show that targeting with EGF_pep‐Au NP‐Pc 4 improves accumulation of fluorescence of Pc 4 in subcutaneous tumors by greater than threefold compared with untargeted Au NPs. Targeted drug delivery and treatment success can be imaged via the intrinsic fluorescence of the PDT drug Pc 4. Using Pc 4 fluorescence, it is demonstrated in vivo that EGF_pep‐Au NP‐Pc 4 impacts biodistribution of the NPs by decreasing the initial uptake by the reticuloendothelial system (RES) and by increasing the amount of Au NPs circulating in the blood 4 h after IV injection. Interestingly, in vivo PDT with EGF_pep‐Au NP‐Pc 4 results in interrupted tumor growth when compared with EGF_pep‐Au NP control mice when selectively activated with light. These data demonstrate that EGF_pep‐Au NP‐Pc 4 utilizes cancer‐specific biomarkers to improve drug delivery and therapeutic efficacy over untargeted drug delivery.     

Therapeutic Pretargeting with Gold Nanoparticles as Drug Candidates for Boron Neutron Capture Therapy

Boron neutron capture therapy (BNCT) is a binary approach for cancer treatment in which boron-10 atoms and thermal neutrons need to colocalize to become effective. Recent research in the development of BNCT drug candidates focuses increasingly on nanomaterials, with the advantages of high boron loadings and passive targeting due to the enhanced permeability and retention (EPR) effect. The use of small boron-rich gold nanoparticles (AuNPs) in combination with a pretargeting approach is proposed. Small sized polyethylene glycol–stabilized AuNPs (core size 4.1 ± 1.5 nm), are synthesized and functionalized with thiolated cobalt bis(dicarbollide) and tetrazine. To enable in vivo tracking of the AuNPs by positron emission tomography (PET), the core is doped with [⁶⁴Cu]CuCl₂. For the pretargeting approach, the monoclonal antibody Trastuzumab is functionalized with trans-cyclooctene-N-hydroxysuccinimide ester. After proving in vitro occurrence of the antibody conjugation onto the AuNPs by click reaction and the low toxicity of the AuNPs, the boron delivery system is evaluated in vivo using breast cancer xenograft bearing mice and PET imaging. Tumor uptake due to the EPR effect can be witnessed with ≈5% injected dose (ID) cm⁻³ at 24 h postinjection, but with slower clearance than expected. Therefore, no increased retention can be observed using the pretargeting strategy.     

The Cellular Interactions of PEGylated Gold Nanoparticles: Effect of PEGylation on Cellular Uptake and Cytotoxicity

Poly(ethylene glycol) (PEG) is frequently used to coat various medical nanoparticles (NPs). As PEG is known to minimize NP interactions with biological specimens, the question remains whether PEGylated NPs are intrinsically less toxic or whether this is caused by reduced NP uptake. In the present work, the effect of gold NP PEGylation on uptake by three cell types is compared and evaluated the effect on cell viability, oxidative stress, cell morphology, and functionality using a multiparametric methodology. The data reveal that PEGylation affects cellular NP uptake in a cell‐type‐dependent manner and influences toxicity by different mechanisms. At similar intracellular NP numbers, PEGylated NPs are found to yield higher levels of cell death, mostly by induction of oxidative stress. These findings reveal that PEGylation significantly reduces NP uptake, but that at similar functional (= cell‐associated) NP levels, non‐PEGylated NPs are better tolerated by the cells.     

Inkjet-Based Technology for Microencapsulation of Gold Nanoparticles within Biocompatible Hydrogels

Herein, an inkjet-based technology as a versatile high throughput methodology for the microencapsulation of gold nanoparticles (AuNPs) inside a biocompatible chitosan hydrogel is described. This continuous automated inkjet production approach generates 30 µm diameter polymeric microcapsules and offers a high rate of production and nanoparticle encapsulation efficiency of 14 nm diameter AuNPs, precise control of the microcapsule size, and ease of scale-up. The hybrid microcapsules demonstrate biocompatible cell-adhesion properties and resist degradation over a large range of pH, making them particularly relevant for a variety of potential health applications.     

Silver Enhancement of Gold Nanoparticles for Biosensing: From Qualitative to Quantitative

Abstract

The establishment and rapid progress in sensitive biosensing using immunogold silver enhancement has occurred in the past 30 years. Its high sensitivity and simplicity have made immunogold silver enhancement a revolutionary technique for signal amplification in biosensing. This review focuses on the major applications of immunogold silver enhancement, with special emphasis on quantitative biosensing. In this review, the combinations of immunogold silver enhancement with a series of quantitative techniques, such as colorimetry, electrical and electrochemical methods, gravimetry, chemiluminescence, Raman spectroscopy, and inductively coupled plasma–mass spectrometry (ICP-MS), are discussed in detail. Immunogold silver enhancement has become one of the most useful methods in highly sensitive quantitative bioanalysis. The recent development of ICP-MS detection shows great potential in combination with immunogold silver enhancement.     

Functionalized Biomimetic Magnetic Nanoparticles as Effective Nanocarriers for Targeted Chemotherapy

Novel MamC‐mediated biomimetic magnetic nanoparticles (BMNPs) are proposed as valuable carriers for targeted chemotherapy because of the size (36 ± 12 nm) and of surface properties conferred by MamC coating. They are super‐paramagnetic at room and body temperatures, have a large magnetic moment per particle, mediate hyperthermia, are cytocompatible, and, having a negative surface charge at physiological pH, can be efficiently coupled with DOXOrubicin (DOXO) and a monoclonal antibody (mAb) directed against the human Met/hepatocyte growth factor receptor (overexpressed in many cancers) displaying coupling stability, while releasing DOXO at acidic pH. This release can be enhanced by hyperthermia. The DOXO‐mAb‐BMNPs selectively recognize Met, bind efficiently to Met⁺ tumor cells, and discharge DOXO within their nuclei more efficiently than DOXO‐BMNPs, exerting cytotoxicity. These data represent proof of concept for future in vivo experiments in which the controlled dual targeting (mAb‐mediated and magnetic) approach and combined (chemotherapy and hyperthermia) therapy will be studied.     

金复合纳米微粒的消光特性

基于Mie光散射理论,研究了金壳介质芯的金复合纳米球壳微粒在光散射与吸收中的消光特性.对微粒增大时的多极子特性以及内半径变动时共振峰的移动作了计算和分析,证实了共振峰位置随内外半径比增大而增大的规律.还对纯金纳米微粒的多极子特性作了计算,并讨论了总消光效率中散射和吸收各自的贡献.     

Fluorescence Correlation Spectroscopy of Gold Nanoparticles

Diffusion dynamics of gold nanoparticles (GNPs) was studied by fluorescence correlation spectroscopy (FCS). The fluorescence was studied by exciting the particles by green laser (532 nm), which is far from longitudinal plasmon band of nanorods. Transmission electron microscope (TEM) and UV-Vis-NIR spectrometer were used to characterize the gold nanoparticles. Despite their low quantum yields, GNPs possess the native fluorescence. The excellent antiphotobleaching behavior of gold nanorods leads to prospects of using FCS for its detailed studies. Using FCS, dynamic information can be extracted from the fluorescence fluctuations in the system by autocorrelation function. Maximum entropy method (MEMFCS) was used to identify the number of distinct components present in the system. The particle sizes obtained from FCS were found to be higher (by few orders of magnitude) compared to TEM analysis. This might be due to the possible contributions from cetyltrimethyl ammonium bromide (CTAB) capping in the system.     

Facile Functionalization of Gold Nanoparticles with PLGA Polymer Brushes and Efficient Encapsulation into PLGA Nanoparticles: Toward Spatially Precise Bioimaging of Polymeric Nanoparticles

Nanocarriers prepared from poly(lactide‐co‐glycolide) (PLGA) have broad biomedical applications. Understanding their cellular uptake and distribution requires appropriate visualization in complex biological compartments with high spatial resolution, which cannot be offered by traditional imaging techniques based on fluorescent or radioactive probes. Herein, the encapsulation of gold nanoparticles (GNPs) into PLGA nanoparticles is proposed, which should allow precise spatial visualization in cells using electron microscopy. Available protocols for encapsulating GNPs into polymeric matrices are limited and associated with colloidal instability and low encapsulation efficiency. In this report, the following are described: 1) a facile protocol to functionalize GNPs with PLGA polymer followed by 2) encapsulation of the prepared PLGA‐capped GNPs into PLGA nanocarriers with 100% encapsulation efficiency. The remarkable encapsulation of PLGA‐GNPs into PLGA matrix obeys the general rule in chemistry “like dissolves like” as evident from poor encapsulation of GNPs capped with other polymers. Moreover, it is shown that how the encapsulated gold nanoparticles serve as nanoprobes to visualize PLGA polymeric hosts inside cancer cells at the spatial resolution of the electron microscope. The described methods should be applicable to a wide range of inorganic nanoprobes and provide a new method of labeling pharmaceutical polymeric nanocarriers to understand their biological fate at high spatial resolution.     

Extremophilic Polysaccharide for Biosynthesis and Passivation of Gold Nanoparticles and Photothermal Ablation of Cancer Cells

Extremophiles are the group of organisms that are far overlooked for exploring novel biomaterials in the field of material science and bionanotechnology. Extremophilic bacterial‐sulfated exopolysaccharide, mauran (MR), is employed for the bioreduction and passivation of gold nanoparticles (AuNps) to enhance the biocompatibility of AuNps and used for photothermal ablation of cancer cells. Here, various concentrations of MR solution are tested for the reduction of HAuCl₄ solution in the presence as well as in the absence of an external reducing agent, to produce mauran‐gold nanoparticles (MRAu Nps). These biocompatible nanocomposites are treated with cancer cell lines under in vitro conditions and NIR irradiated for complete ablation. MRAu Nps‐treated cancer cells on immediate exposure to infrared radiation from a femtosecond pulse laser of operating wavelength 800 nm are subjected to hyperthermia causing cell death. Biocompatible MR stabilization could fairly reduce the cytotoxicity caused by bare AuNps during biomedical applications. Application of a biocompatible polysaccharide from extremophilic bacterial origin for reduction and passivation of AuNps and used for a biomedical purpose is known to be first of its kind in bionanofusion studies.     

Biological Production of Gold Nanoparticles at Different Temperatures: Efficiency Assessment

The study aims to compare different approaches and efficacies during the biological production of nanoparticles (NPs). Gold nanoparticles (AuNPs) are produced by Fusarium oxysporum at two different temperatures. One flask is incubated at 37 °C (“Common”) and the other is directly heated for 5 min at 80 °C (“Heat-treated”). Obtained AuNPs are analyzed and compared by spectrophotometry, transmission electron microscopy (TEM), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), dynamic light scattering (DLS), and Fourier transform infrared spectroscopy (FTIR). Graphite furnace atomic absorption spectroscopy (GF-AAS) is used to determine the particle concentration after the AuNPs production. The AuNPs prepared by both (the Common and the Heat-treated) methods exhibit maximum absorption peaks at 541 and 528 nm, respectively, and have round shapes and sizes of less than 50 nm. Their zeta potential is about −28 mV. GF-AAS shows that the efficiency of AuNP production in Common- and Heat-treated samples is equal, between 65% and 68%. Since the Heat-treated sample shows a better size distribution, the use of higher temperature and shorter time period is preferable for the bioproduction of AuNPs. It seems that shortening the time for the production of AuNPs prevents the formation of larger NPs.     

Influence of Morphological Homogeneity of Superspherical Gold Nanoparticles on Plasmonic Photothermal Heat Generation

The plasmonic photothermal (PPT) characteristics of gold nanostructures have been extensively investigated theoretically and experimentally due to their potential for use materials science and industry. The management of the size and shape of gold nanoparticles has been a key issue in the development of better solutions for PPT heat generation because their size and shape determine their resultant photothermal properties. However, the light absorption of gold nanostructures is mainly dependent on the wavelength and orientation of the incident light; hence, maintaining uniform size and shape is critical for achieving maximum photothermal energy. Morphologically homogeneous spherical gold nanoparticles, or super gold nanospheres prepared by slowly etching uniform octahedral gold nanoparticles, demonstrate better PPT heat generation compared with commercially available nonsmooth gold nanoparticles (GNSs). The PPT heating experiments show a maximum temperature difference of 5.7 °C between the super and ordinary GNSs with the same average maximum Feret's diameters, which result from the more efficient PPT heat power generation (20.6%) of the super GNSs. In an electromagnetic‐wave simulation, the super GNSs show lower polarization dependence and a 24.6% higher absorption cross‐section than ordinary GNSs.     

Size Controlled Synthesis of Gold Nanoparticles using Photochemically Prepared Seed Particles

Gold nanoparticles having prechosen size ranging from 5 to 110 nm have been prepared in two steps. Firstly, small spherical particles (seed) of average diameters between 5 and 20 nm were prepared by varying the ratio of gold ion concentration to stabilizer/reductant, TX-100 concentration and using UV irradiation. Secondly, 20–110 nm particles were formed by a non-iterative seed-mediated growth where small particles produced by the above irradiation technique were exploited as seeds and fresh Au(III) ions were reduced onto the surface on the seed particles by ascorbic acid. The kinetics of particle formation has also been reported. These methods were fast and showed improved monodispersity sphericity and excellent reproducibility.     

Nucleation of Polypropylene with Gold Nanoparticles. Part 1: Introduction of Sandwich Method for Evaluation of Very Weak Nucleation Activity

Although gold particles are known to nucleate isotactic polypropylene (PP), the nucleating effect of chemically pure 5 nm Au, prepared in vacuum sputter coater, was found to be hardly observable. In order to detect such a weak effect, we deposited a homogeneous layer of Au nanoparticles between thin PP films and evaluated the nucleation activity by a combination of three independent methods: polarized light microscopy (PLM), differential scanning calorimetry (DSC), and 2D wide-angle X-ray scattering (2D-WAXS). This new technique, which was called sandwich method, allowed us to demonstrate that gold nanoparticles were able to nucleate PP crystallization, although the effect was much weaker than that produced by commercial α-nucleant [1,2,3,4-bis(3,4-dimethylbenzylidene)sorbitol] and β-nucleant (N,N-Dicyclohexyl-2,6-naphthalene dicarboxamide). The sandwich method appeared to be quite universal and applicable for any micro-sized nucleants or nanonucleants.     

纳米金粒子对裸鼠胃癌组织近红外光谱范围的组织光学特性影响

为明确纳米金粒子对肿瘤组织近红外光学成像的影响,研究观察了肿瘤组织于纳米金粒子贮积后对其近红外光谱范围的光学特性的变化特点.实验中,应用MCG-803人胃腺癌细胞系(来源于CTCC),采用原位接种法建立BALB/C裸小鼠胃癌模型,给予动物静脉注射纳米金粒子悬液,以组织切片形式检测肿瘤组织近红外光衰减系数的特征性变化.结...     

Step-addition Method for Enhancing the Yield of Gold Nanoparticles in Block Copolymer Solution

Triblock copolymer poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) has been used to synthesize gold nanoparticles from hydrogen tetrachloroaureate (III) hydrate (HAuCl₄·3H₂O) salt in aqueous solution at room temperature. Measurements were performed using the triblock copolymer Pluronic P85 (EO₂₆PO₃₉EO₂₆) at a fixed concentration (1 wt%) mixed with varying HAuCl₄·3H₂O concentration in the range of 0.001 to 0.1 wt%. The surface plasmon resonance (SPR) band in UV-visible absorption spectra confirmed the formation of the gold nanoparticles. The maximum yield of the nanoparticles was found at 0.005 wt% of the salt solution. Small-angle neutron scattering (SANS) does not show any significant change in the scattering profile in these suspensions of the nanoparticles. A similar behavior was also observed in dynamic light scattering (DLS) experiments where autocorrelation function was found to be independent of the salt concentration. This can be understood since a high-block copolymer-to-gold ion ratio (r ～ 22) is required in the reduction reaction to produce gold particles. As a result, a very small fraction of the block copolymers were associated with the gold nanoparticles, and hence lead to a very low yield. Both SANS and DLS basically see the micelles of most of these block copolymers, which are not associated with nanoparticles. Based on this explanation, a step-addition method was used to enhance the yield of gold nanoparticles by manifold, where the gold salt is added in small steps to maintain higher value of r (>22), and therefore continuous formation of nanoparticles.     

Assembly of Gold Nanoparticles on Gold Nanorods Using Functionalized Poly(N‐isopropylacrylamide) as Polymeric “Glue”

A telechelic thermoresponsive polymer, α‐amino‐ω‐thiol‐poly(N‐isopropylacrylamide) (H₂N‐PNiPAM‐SH), is used as the polymeric glue to assemble gold nanoparticles (AuNPs) around gold nanorods (AuNRs) into a satellite structure. Prepared by reversible addition‐fragmentation chain transfer polymerization followed by hydrazinolysis, H₂N‐PNiPAM‐SH is able to interlink the two types of the gold building blocks with the thiol‐end grafting on AuNRs and the amine‐end coordinating on the AuNP surface. The density of the grafted AuNPs on AuNRs can be tuned by adjusting the molar ratio between AuNPs and AuNRs in the feed. The resulted satellite‐like assembly exhibits unique optical property that was responsive to temperature change.     

Blood Compatibility of Multilayered Polyelectrolyte Films Containing Immobilized Gold Nanoparticles

Surface material functionalization including layer‐by‐layer (LbL) polyelectrolyte films with incorporated nanoparticles is a growing field with a wide range of biomedical applications: drug reservoirs, medical devices, or tissue engineering. In parallel, gold nanoparticles (AuNPs) can be grafted by drugs and sensitive molecules using simple protocols. This study shows that AuNP behavior is modified when they are entrapped into three partner LbL films in comparison to the colloidal solution. A polycationic (polyallylamine hydrochloride (PAH)) and a polyanionic (polyacrylic acid (PAA)) polymer is used to build films based on three cycles ((PAH/AuNP/PAA)₃). To investigate the interaction with biomolecules and cells, three different films are developed changing the outer layer (either PAH or AuNP or PAA) with the same number of AuNP deposit. The best biocompatibility is observed with a polyacrylic acid outer layer. Due to the high capacity of drug grafting on gold nanoparticles, the results seem promising for the development of nanostructured biomedical devices.