PEG-modified Gold Research Articles

Cellular uptake and cytotoxicity of PEGylated gold nanoparticles in C33A cervical cancer cells

Gold nanoparticles (GNPs) have served as an excellent candidate for biomedical applications. GNPs can be conjugated with carboxyl-polyethylene glycol-thiol (PEG) as a stealth coating which prolongs circulation time [Lipka J et al 2010 Biodistribution of PEG-modified gold nanoparticles following intratracheal instillation and intravenous injection. Biomaterials, 31 , 6574–6581, Janát-Amsbury M et al 2011 Geometry and surface characteristics of gold nanoparticles influence their biodistribution and uptake by macrophages. Eur. J. Pharm. Biopharm, 77 , 417–423] and increases cellular uptake.[He B et al 2017 Increased cellular uptake of peptide-modified PEGylated gold nanoparticles. Biochem. Biophys. Res. Commun., 494 , 339–345, Soenen S. J et al 2014 , The cellular interactions of PEGylated gold nanoparticles: effect of PEGylation on cellular uptake and cytotoxicity. Part. Part. Syst. Charact., 31 , 794–800, Guo J et al 2016 Bioconjugated gold nanoparticles enhance cellular uptake: A proof of concept study for siRNA delivery in prostate cancer cells. Int. J. Pharm., 509 , 16–27. Brandenberger C et al 2010 Quantitative evaluation of cellular uptake and trafficking of plain and polyethylene glycol‐coated gold nanoparticles. Small, 6 , 1669–1678. To examine the biological effects of PEG-coated GNPs, we investigated their cytotoxicity on human cervical cancer C33A cells as compared to citrate-capped GNPs. Our results indicated that PEGylated GNPs markedly induce apoptosis and necrosis causing cell shrinkage and cell membrane asymmetry. 30 nm citrate-capped GNPs were synthesized in aqueous solution using a citrate-reduction method. GNPs were functionalized with PEG (MW = 7500 g mol−1. The GNPs were characterized using scanning electron microscopy (SEM), confirming that the as-synthesized GNPs have a diameter of 30 nm. Dynamic light scattering (DLS) determined that the hydrodynamic diameter of PEGylated GNPs was 78.82 nm, and that of citrate-capped GNPs was 43.82 nm. Zeta potential measurements showed an increase in colloidal stability for PEGylated GNPs as compared to citrate GNPs, with a zeta potential of −33.33 mV observed for citrate-capped GNPs and a zeta potential of −43.38 mV observed for PEGylated GNPs. The PEGylated GNPs were found to effectively induce early and late-stage apoptosis in C33A cells with a significant reduction in total cell viability of 32.3%. Based on the apoptotic activity in C33A cells, PEGylated GNPs may serve as a promising radiosensitizer for cancer treatments.

Intestinal Mucin Induces More Endocytosis but Less Transcytosis of Nanoparticles across Enterocytes by Triggering Nanoclustering and Strengthening the Retrograde Pathway.

Mucus, which is secreted by the goblet cells of enterocytes, constitutes the first obstacle encountered for the intestinal absorption of nanomedicines. For decades, mucus has simply been regarded as a physical barrier that hinders the permeation and absorption of drugs, because of its high viscosity and reticular structure, whereas the interaction of mucus ingredients with nanomedicines is usually neglected. It is unclear whether glycoproteins, as the main components of mucus, interact with nanomedicines. We also do not know how the potential interaction affects the subsequent transportation of nanomedicines through the intestinal epithelium. In this study, mucin as the key element of mucus was investigated to characterize the interaction of nanomedicines with mucus. PEG-modified gold nanoparticles (PGNPs) were fabricated as model nanoparticles. Mucin was found to adhere to the nanoparticle surface to form a corona structure and induce the clustering of PGNPs by joining particles together, demonstrating the interaction between mucin and PGNPs. In addition, two intestinal epithelia, Caco-2 (non- mucus secretion) and HT-29 (high mucus secretion), were compared to evaluate the influence of mucin on the cellular interaction of PGNPs. Amazingly, mucin altered the trafficking characteristic of PGNPs in intestinal epithelium. Both in vitro and in vivo investigations demonstrated more nanoparticles being internalized by cells due to the mucin coverage. However, mucin induced a significant reduction in the transcytosis of PGNPs across epithelial monolayers. The mechanism exploration further revealed that the "more endocytosis but less transcytosis (MELT)" effect was mainly attributed to the strengthened retrograde pathway in which more PGNPs were transported to Golgi apparatus and exocytosed back to the apical but not the basolateral side of the epithelial monolayers. The "MELT" effect endowed mucin with duality in the nanoparticle transportation. Therefore, the rational regulation based on the "MELT" effect will provide new insight into overcoming the mucus obstacle as a barrier and enhancing the oral absorption rate of nanomedicines.

Surface-Enhanced Raman Scattering from Self-Assembled Film of Thiolated Peg-Modified Gold Nanoparticles

Self-assembly of thiolated polyethylene glycol (PEG)-modifi ed Au nanoparticles (NPs) into a closely packed particle film at a water–air interface for the production of reproducible, highly active surface-enhanced Raman scattering (SERS) was developed. Upon modifi cation with thiolated PEG, the Au NPs can sustain severe conditions (e.g., 3 M NaCl solution), and after separation from colloids and drying, they can be redispersed into water, forming colloids again. We found that the thiolated PEG-modifi ed Au NPs, from their concentrated colloids, can spontaneously selfassemble into a two-dimensional (2D) closely packed particle film at the water–air interface. With abundant hot spots created and exposed, the 2D particle film produces large, reproducible SERS signal for 4-mercaptobenzoic acid (probe molecule), reaching a detection limit of 1 × 10–11 M. These results offer a useful way for the storage of Au NPs and the fabrication of highly reproducible and active SERS substrate.

Nd:YAG laser-induced morphology change and photothermal conversion of gold nanorods with potential application in the treatment of port-wine stain.

Based on the principle of selective photothermolysis, 1064nm Nd:YAG laser has great potential for the treatment of deeper and larger PWS. However, the clinical effectiveness is limited because of the weak absorption of blood to Nd:YAG laser. The aim of this study is to obtain the optimal irradiation conditions to effectively destroy vascular lesions with the assistance of PEG-modified gold NRs to enhance blood absorption of Nd:YAG laser. In our study, PEG-modified gold NRs were prepared by the seeded growth method. Gold NRs after exposure to Nd:YAG laser were characterized using absorption spectra and transmission electron microscope images. The tissue-like phantom containing a glass capillary with blood was prepared and exposed to Nd:YAG laser to investigate the laser energy density and pulse number required for blood coagulation before and after the addition of gold NRs in blood. The results show that the millisecond Nd:YAG laser irradiation does not result in the shape change of gold NRs. After injection of gold NRs into the bloodstream (4.60mg/kg), the absorbance of blood at 1064nm increased 3.9 times. The threshold energy density for the treatment of PWS decreased by 33% (from 30 to 20J/cm2). Our findings provide an experimental guide for choosing laser parameters and gold NRs concentration for the treatment of deeper and larger PWS with the assistance of PEG-modified gold NRs in vivo in the future.

Enhancement of light absorption by blood to Nd:YAG laser using PEG-modified gold nanorods.

On the basis of the principle of selective photothermolysis, laser therapy has been the most effective treatment strategy for Port-wine stains (PWSs) caused by the expansion of dermal capillaries. Neodymium:Yttrium Aluminum Garnet (Nd:YAG) laser at 1064 nm wavelength has great potential for deeply buried PWS, although its application is limited because of its weak absorption by blood. The purpose of this study is to investigate the effect of PEG-modified gold nanorods (NRs) on the blood absorption enhancement for Nd:YAG laser. PEG-modified gold nanorods (NRs) were synthesized via the seeded growth method. Then, the effect of PEG-modified gold NRs on blood light absorbance was investigated through adding different concentration of PEG-modified gold NRs to 1 ml of blood at room temperature. Finally, the optical properties of whole mice blood with or without PEG-modified gold NRs under slow heating were investigated. The average length and width of PEG-modified gold NRs are 79.5 ± 10.5 and 13.5 ± 0.9 nm, respectively, with the aspect ratio of 5.89, and a strong absorption peak exists at ∼1050 nm in the near-infrared range. A linear correlation between the blood absorbance at 1064 nm and the amount of PEG-modified gold NRs was obtained. The absorbance at 1064 nm increased 17.6, 33.0, 48.3, and 65.4 times when 0.4, 0.8, 1.2, and 1.6 mg of PEG-modified gold NRs was added to 1 ml of blood at room temperature, respectively. After adding 0.8 mg of PEG-modified gold NRs to 1 ml of blood, blood absorbance at 1064 nm at different temperatures increased by an average of 24.0 times. After intravenously injecting PEG-modified gold NRs (0.87 mg/ml) into Sprague-Dawley mice, the blood absorbance at 1064 nm increased from 0.014 to 0.5. Our findings suggest that PEG-modified gold NRs injection is an efficient way to enhance light absorption by blood to Nd:YAG laser. Lasers Surg. Med. 48:790-803, 2016. © 2016 Wiley Periodicals, Inc.

Supramolecular hybrid hydrogel based on host-guest interaction and its application in drug delivery.

In this work, we developed a simple, novel method for constructing gold nanocomposite supramolecular hybrid hydrogels for drug delivery, in which gold nanocrystals were utilized as building blocks. First, methoxypoly(ethylene glycol) thiol (mPEG-SH, molecular weight (MW)=5 K) capped gold nanocrystals (nanospheres and nanorods) were prepared via a facile one-step ligand-exchange procedure. Then, the homogeneous supramolecular hybrid hydrogels were formed, after adding α-cyclodextrin (α-CD) into PEG-modified gold nanocrystal solutions, due to the host-guest inclusion. Both gold nanoparticles and inclusion complexes formed between α-CD and PEG chain provided the supra-cross-links, which are beneficial to the gelation formation. The resulting hybrid hydrogels were fully characterized by a combination of techniques including X-ray diffraction, rheology studies, and scanning electron microscopy. Meanwhile, the hybrid hydrogel systems demonstrated unique reversible gel-sol transition properties at a certain temperature caused by the temperature-responsive reversible supramolecular assembly. The drug delivery applications of such hybrid hydrogels were further investigated in which doxorubicin was selected as a model drug for in vitro release, cytotoxicity, and intracellular release studies. We believe that the development of such hybrid hydrogels will provide new and therapeutically useful means for medical applications.

Fast loading of PEG–SH on CTAB-protected gold nanorods

This study aimed to develop a rapid and simple method to functionalize gold nanorods using thiolated polyethylene glycol (PEG–SH), which can be used for plasmonic sensing and other applications. PEG was successfully loaded onto CTAB-protected gold nanorods in 30 min using Tris buffer with a pH of 3.0. In contrast, the functionalization of gold nanorods in water induced the aggregation of gold nanorods and produced a floating layer of randomly aggregated nanorods at the water–air interface. The concentration of mPEG–SH has no significant influence on the optical properties of gold nanorods modified by the proposed method. The results show that the PEG-modified gold nanorods could sustain 1.0 M NaCl, whereas CTAB-protected gold nanorods aggregated immediately after the addition of NaCl. Compared with the traditional 24 h method in a CTAB/carbonate solution, the proposed method offers a higher loading speed and a higher coverage density of PEG on gold nanorods. It was also found that the PEG-modified NR-610 could form a very uniform monolayer on silicon by slow evaporation of the water content. This method may open a new route for fabricating substrates for bio-medication, detection of food contaminants, and environmental monitoring.

The effect of PEG-5K grafting level and particle size on tumor accumulation and cellular uptake

PEG-modified gold nanoparticles (PEG-modified GNs) with diameters of 40 nm and 70 nm were prepared to elucidate the effect of extent of PEG (M.W. 5000) grafting and particle size on tumor accumulation and cellular uptake. Flow cytometry reveals that cellular uptake is strongly related to the size of PEG-modified GNs, rather than the extent of PEG-5K grafting level. Cytotoxicity analysis based on the intracellular release of drugs showed that the 70 nm PEG-modified GNs have the higher cytotoxicity, beccause of their greater cellular uptake. Also, particle size, rather than PEG-5K grafting level affects tumor accumulation. However, PEG-5K grafting level significantly affects the accumulation of particles in the liver and spleen. This finding is important in determining the proper PEG-5K grafting level and particle size for designing nano-medicines.

12P-Conjugated PEG-Modified Gold Nanorods Combined with Near-Infrared Laser for Tumor Targeting and Photothermal Therapy

Gold nanorods have been reported as potential tumor photothermal therapy in vivo and in vitro. However, development of the safe and efficient tumor-targeting gold nanorods for in vivo localized tumor therapy is still a challenge. In our present study, we synthesized the PEG modified gold nanorods and demonstrated its negligible cytotoxicity in vitro. These nanorods also have been demonstrated to efficiently ablate the different kinds of tumor cells in vitro after exposure to the near-infrared laser. When the PEG modified gold nanorods conjugated with the 12P (sequence: TACHQHVRMVRP), this conjugate showed great tumor-targeting and hyperthermia effects on the human liver cancer cell line HepG2 in vitro when coupled with the near-infrared laser treatment. To determine the potential hyperthermia effect of PEG modified gold nanorods or 12P conjugate on tumor cells in vivo, the mice hepatic cancer cells were used to induce the subcutaneous tumor-bearing model in ICR mice. The significant inhibition effects of near-infrared laser mediated PEG modified gold nanorods or 12P conjugate on the tumor growth were observed. These composite results suggest that the 12P-conjugated PEG modified gold nanorods exhibit great biocompatible, particular tumor-targeting and effective photothermal ablation of tumor cells, which warrant the potential therapeutic value of this conjugate for further application in in vivo localized tumor therapy.

Biodistribution of PEG-modified gold nanoparticles following intratracheal instillation and intravenous injection

Besides toxicity tests, biokinetic studies are a fundamental part of investigations to evaluate a safe and sustainable use of nanoparticles. Today, gold nanoparticles (Au NPs) are known to be a versatile tool in different areas such as science, engineering or medicine. In this study, we investigated the biokinetics after intravenous and intratracheal applications of poly(ethylene glycol) (PEG) modified Au NPs compared to plain Au NPs. Radioactive-labeled Au NPs of 5 nm inorganic core diameter were applied to rats and the NP content in tissues, organs and excretion were quantified after 1-hour and 24-hours. After intravenous injection, a prolonged blood circulation time was determined for Au NPs with 10 kDa PEG chains. Non-PEGylated Au NPs and 750 Da PEG Au NPs accumulated mostly in liver and spleen. After intratracheal application the majority of all three types of applied NPs stayed in the lungs: the total translocation towards the circulation did not differ considerably after PEGylation of the Au NPs. However, a prolonged retention time in the circulation was detected for the small fraction of translocated 10 kDa PEG Au NPs, too.

Development of functional gold nanorods for bioimaging and photothermal therapy

Gold nanorods have strong surface plasmon band at near-infrared light region, and are used as a photothermal converter. Since the near-infrared light penetrates into tissues deeply, it has been expected as a contrast agent for near infrared light bioimaging, a photosensitizer for photothermal therapy, and functional device for drug delivery system responding to near-infrared light irradiation. In this study, the surface plasmon bands of intravenously injected gold nanorods were monitored in the mouse abdomen using a spectrophotometer equipped with an integrating sphere, then we determined pharmacokinetics parameters of the gold nanorods after intravenous injection. Next, the PEG-modified gold nanorods were directly injected into subcutaneous tumors in mice, then, near-infrared pulsed laser light was irradiated the tumors. Significant tumor damage and suppression of the tumor growth was observed. We constructed targeted delivery system of the gold nanorods by modifying with a thermo-responsive polymer and a peptide responding to a protease activity. These modified gold nanorods are expected as functional nanodevices for photothermal therapy and drug delivery system.

Nuclear penetration of surface functionalized gold nanoparticles

Free gold nanoparticles easily aggregate when the environment conditions change. Here, gold nanoparticles (AuNPs) with average diameter of 3.7 nm were prepared and then modified with poly(ethylene glycol) (PEG) to improve stability. The gold nanoparticles were first surface-modified with 3-mercaptopropionic acid (MPA) to form a self-assembled monolayer and subsequently conjugated with NH 2-PEG-NH 2 through amidation between the amine end groups on PEG and the carboxylic acid groups on the particles. The biocompatibility and intracellular fate of PEG-modified gold nanoparticles (AuNP@MPA-PEG) were then studied in human cervical cancer (HeLa) cells. Cell viability test showed that AuNP@MPA-PEG did not induce obvious cytotoxicity. Both confocal laser scanning microscopy and transmission electron microscopy demonstrated that AuNP@MPA-PEG entered into mammalian cells and the cellular uptake of AuNP@MPA-PEG was time-dependent. Inductively coupled plasma mass spectrometry and confocal microscopy imaging further demonstrated that AuNP@MPA-PEG penetrated into the nucleus of mammalian cells upon exposure for 24 h. These results suggest that surface modification can enhance the stability and improve the biocompatibility. This study also indicates that AuNP@MPA-PEG can be used as potential nuclear targeted drug delivery carrier.

Functional Controlled Release Systems Triggered by Photothermal Effect of Gold Nanorods

AbstractGold nanorods, rod-shaped gold nanoparticles, have transverse and longitudinal surface plasmon (SP) bands at visible and near-infrared (IR) regions, respectively. Since the absorbed light energy is converted into heat, photothermal effect of gold nanorods can be triggered without damaging the tissues in the path of near-IR laser light. In this study, we tried to construct controlled release system of functional molecules from surface of gold nanorods mediated by the photothermal effect. First, we evaluated controlled release of poly(ethyleneglycol) (PEG) chains from PEG-modified gold nanorods (PEG-NR). Next, we employed double stranded oligonucleotide as a thermo-responsive dissociating group (DNA-NR). Finally, we evaluated photothermal release of PEG chains mediated retro-Diels-Alder reaction (PEG-DA-NR). For construction of controlled release system of functional molecules, these studies will provide important information about the photothermal reactions of surface molecules on the gold nanorods triggered by near-IR light irradiation.

In vivo Monitoring of Gold Nanorods and Tissue Damage Mediated with Their Photothermal Effect

AbstractGold nanorods have a strong surface plasmon band at the near infrared region. The absorbed light energy is then converted to heat. Since near infrared light can penetrate deeply into tissue, gold nanorods are expected to be used as a contrast agent for bioimaging using the near infrared light and photosensitizers for photothermal therapy. The surface plasmon bands of intravenously injected the gold nanorods were directly monitored from the mouse abdomen by using a spectrophotometer equipped with an integrating sphere. The absorbance at 900 nm from PEG5,000-modified gold nanorods immediately increased after injection and reached a plateau. The injection of phosphatidylcholine-modified gold nanorods also increased the absorbance at 900 nm, but the absorbance decreased single exponentially with a 1.3-min half-life. To demonstrate photothermal tumor therapy, the PEG-modified gold nanorods were directly injected into subcutaneous tumors in mice, then, near infrared laser light was irradiated to the tumor. After the treatment, significant suppression of tumor growth was observed.

Application of PEG-modified Gold Nanorods to Bioscience Field

Application of PEG-modified Gold Nanorods to Bioscience Field

PEG-modified gold nanorods with a stealth character for in vivo applications

Gold nanorods prepared in hexadecyltrimethylammonium bromide (CTAB) solution are expected to provide novel materials for photothermal therapy and photo-controlled drug delivery systems. Since gold nanorods stabilized with CTAB show strong cytotoxicity, we developed a technique to modify these with polyethyleneglycol (PEG) for medical applications. PEG-modification was achieved by adding mPEG-SH in the CTAB solution, then, excess CTAB was removed by dialysis. PEG-modified gold nanoparticles showed a nearly neutral surface, and had little cytotoxicity in vitro. Following intravenous injection into mice, 54% of injected PEG-modified gold nanoparticles were found in blood at 0.5 h after intravenous injection, whereas most of gold was detected in the liver in the case of original gold nanorods stabilized with CTAB.