Solvent vapor annealing of block copolymers in confined topographies: commensurability considerations for nanolithography.

The directed self-assembly of block copolymer (BCP) materials in topographically patterned substrates (i.e., graphoepitaxy) is a potential methodology for the continued scaling of nanoelectronic device technologies. In this Communication, an unusual feature size variation in BCP nanodomains under confinement with graphoepitaxially aligned cylinder-forming poly(styrene)-block-poly(4-vinylpyridine) (PS-b-P4VP) BCP is reported. Graphoepitaxy of PS-b-P4VP BCP line patterns (CII ) is accomplished via topo-graphy in hydrogen silsequioxane (HSQ) modified substrates and solvent vapor annealing (SVA). Interestingly, reduced domain sizes in features close to the HSQ guiding features are observed. The feature size reduction is evident after inclusion of alumina into the P4VP domains followed by pattern transfer to the silicon substrate. It is suggested that this nano-domain size perturbation is due to solvent swelling effects during SVA. It is proposed that using a commensurability value close to the solvent vapor annealed periodicity will alleviate this issue leading to uniform nanofins.

Swelling of ionic and nonionic surfactant micelles by high pressure gases.

The influence of different solvent environments on the size, shape, and characteristics of surfactant micelles of Pluronic F127 and CTAB was investigated by small-angle neutron scattering (SANS). SANS experiments were undertaken on dilute micellar surfactant solutions of F127 and CTAB that between them were exposed to liquid and supercritical carbon dioxide, liquid propane, ethane, and heptane under various pressures and temperatures. Swelling of the surfactant micelles could be directly related to the solubility of the solvents within the micelles, especially within their cores. Carbon dioxide produced the largest swelling of the Pluronic F127 micelles, compared to propane and ethane, which mirrors the solubility of the gases in the PPO core of the micelles. Conversely, the extent of swelling of the cores of CTAB micelles was greater with propane compared to carbon dioxide, which again relates to the solubility of the solvents in the alkane core of the CTAB micelles.

Chemical interactions and their role in the microphase separation of block copolymer thin films.

The thermodynamics of self-assembling systems are discussed in terms of the chemical interactions and the intermolecular forces between species. It is clear that there are both theoretical and practical limitations on the dimensions and the structural regularity of these systems. These considerations are made with reference to the microphase separation that occurs in block copolymer (BCP) systems. BCP systems self-assemble via a thermodynamic driven process where chemical dis-affinity between the blocks driving them part is balanced by a restorative force deriving from the chemical bond between the blocks. These systems are attracting much interest because of their possible role in nanoelectronic fabrication. This form of self-assembly can obtain highly regular nanopatterns in certain circumstances where the orientation and alignment of chemically distinct blocks can be guided through molecular interactions between the polymer and the surrounding interfaces. However, for this to be possible, great care must be taken to properly engineer the interactions between the surfaces and the polymer blocks. The optimum methods of structure directing are chemical pre-patterning (defining regions on the substrate of different chemistry) and graphoepitaxy (topographical alignment) but both centre on generating alignment through favourable chemical interactions. As in all self-assembling systems, the problems of defect formation must be considered and the origin of defects in these systems is explored. It is argued that in these nanostructures equilibrium defects are relatively few and largely originate from kinetic effects arising during film growth. Many defects also arise from the confinement of the systems when they are 'directed' by topography. The potential applications of these materials in electronics are discussed.

Development of anisamide-targeted PEGylated gold nanorods to deliver epirubicin for chemo-photothermal therapy in tumor-bearing mice.

BACKGROUND: Gold nanorods (AuNRs), due to the optical and electronic properties namely the surface plasma resonance, have been developed to achieve the light-mediated photothermal therapy (PTT) for cancer. However, PTT alone may suffer from inefficient tumor killing. Recently, the combination of PTT and chemotherapy has been utilized to achieve synergistic anticancer effects. METHODS: In this study, AuNRs capped with hexadecyltrimethylammonium bromide (CTAB), poly(acrylic acid) (PAA), and PEGylated anisamide (a ligand known to target the sigma receptor) have been developed to produce a range of negatively charged anisamide-targeted PEGylated AuNRs (namely Au-CTAB-PAA-PEG-AA) for the combination of PTT and chemotherapy (termed as chemo-photothermal therapy [CPTT]). Epirubicin (EPI, an anthracycline drug) was efficiently loaded onto the surface of Au800-CTAB-PAA-PEG-AA via the electrostatic interaction forming Au800-CTAB-PAA-PEG-AA.EPI complex. RESULTS: The resultant complex demonstrated pH-dependent drug release, facilitated nucleus trafficking of EPI, and induced antiproliferative effects in human prostate cancer PC-3 cells. When Au800-CTAB-PAA-PEG-AA.EPI complex was further stimulated with desired laser irradiation, the synergistic outcome was evident in PC-3 xenograft mice. CONCLUSION: These results demonstrate a promising strategy for clinical application of CPTT in cancer.

Anisamide-targeted PEGylated gold nanoparticles designed to target prostate cancer mediate: Enhanced systemic exposure of siRNA, tumour growth suppression and a synergistic therapeutic response in combination with paclitaxel in mice.

Small interfering RNA (siRNA) has recently illustrated therapeutic potential for malignant disorders. However, the clinical application of siRNA-based therapeutics is significantly retarded by the paucity of successful delivery systems. Recently, multifunctional gold nanoparticles (AuNPs) as non-viral delivery carriers have shown promise for transporting chemotherapeutics, proteins/peptides, and genes. In this study, AuNPs capped with polyethylenimine (PEI) and PEGylated anisamide (a ligand known to target the sigma receptor) have been developed to produce a range of positively charged anisamide-targeted PEGylated AuNPs (namely Au-PEI-PEG-AA). The anisamide-targeted AuNPs effectively complexed siRNA via electrostatic interaction, and the resultant complex (Au110-PEI-PEG5000-AA.siRNA) illustrated favourable physicochemical characteristics, including particle size, surface charge, and stability. In vitro, anisamide-targeted AuNPs selectively bound to human prostate cancer PC-3 cells, inducing efficient endosomal escape of siRNA, and effective downregulation of the RelA gene. In vivo, prolonged systemic exposure of siRNA was achieved by anisamide-targeted AuNPs resulting in significant tumour growth suppression in a PC3 xenograft mouse model without an increase in toxicity. In addition, a combination of siRNA-mediated NF-κB knockdown using anisamide-targeted AuNPs with Paclitaxel produced a synergistic therapeutic response, thus providing a promising therapeutic strategy for the treatment of prostate cancer.

Nanopatterned protein-polysaccharide thin films by humidity regulated phase separation.

Greater sustainability in mass manufacturing is essential to alleviating anthropogenic climate change. High surface-area, micro- and nano-patterned films have become a fundamental tool in materials science, however these technologies are subject to a dwindling petrochemical supply, increasing costs and disposability concerns. This paper describes the production of patterned biopolymer films utilizing controlled phase separation of biopolymeric thin films into nanopatterns using easily transferable variables and methods. Similar morphologies to those commonly observed with synthetic block-copolymers (BCPs) were achieved across a large range of feature sizes, from 160 nm to >5 µm: Bicontinuous, porous, droplet-matrix, particulated and dimpled. Protein and polysaccharide type, protein to polysaccharide ratio, casting method and ambient humidity were primary conditions found to influence the pore morphology of the films. High protein concentrations (4:1 and 2:1 blends) generally resulted in porous structures whereas high polysaccharide concentrations (1:2 and 1:4 blends) resulted in spherical structures. High humidity conditions (60% + relative humidity) resulted in the growth of large protuberances up to 10 µm in diameter while lower humidity (10-30%) resulted in discrete features smaller than 200 nm.

Bioconjugated gold nanoparticles enhance cellular uptake: A proof of concept study for siRNA delivery in prostate cancer cells.

The chemistry of gold nanoparticles (AuNPs) facilitates surface modifications and thus these bioengineered NPs have been investigated as a means of delivering a variety of therapeutic cargos to treat cancer. In this study we have developed AuNPs conjugated with targeting ligands to enhance cell-specific uptake in prostate cancer cells, with a purpose of providing efficient non-viral gene delivery systems in the treatment of prostate cancer. As a consequence, two novel AuNPs were synthesised namely AuNPs-PEG-Tf (negatively charged AuNPs with the transferrin targeting ligands) and AuNPs-PEI-FA (positively charged AuNPs with the folate-receptor targeting ligands). Both bioconjugated AuNPs demonstrated low cytotoxicity in prostate cancer cells. The attachment of the targeting ligand Tf to AuNPs successfully achieved receptor-mediated cellular uptake in PC-3 cells, a prostate cancer cell line highly expressing Tf receptors. The AuNPs-PEI-FA effectively complexed small interfering RNA (siRNA) through electrostatic interaction. At the cellular level the AuNPs-PEI-FA specifically delivered siRNA into LNCaP cells, a prostate cancer cell line overexpressing prostate specific membrane antigen (PSMA, exhibits a hydrolase enzymic activity with a folate substrate). Following endolysosomal escape the AuNPs-PEI-FA.siRNA formulation produced enhanced endogenous gene silencing compared to the non-targeted formulation. Our results suggest both formulations have potential as non-viral gene delivery vectors in the treatment of prostate cancer.

Evaluation of the physicochemical properties and the biocompatibility of polyethylene glycol-conjugated gold nanoparticles: A formulation strategy for siRNA delivery.

The potential of RNA interference (RNAi)-based therapeutics for cancer has received much attention; however, delivery of RNAi effectors, such as small interfering RNA (siRNA), remains an obstacle to clinical translation. Non-viral delivery vectors have been used extensively to enhance siRNA delivery. Recently, the potential of gold nanoparticles (AuNPs) for transporting drugs, proteins and genetic materials has been demonstrated. Previously, our laboratory synthesised positively charged, surfactant-free AuNPs in water by the reduction of gold (III) chloride (AuCl3) using hydroxylamine hydrochloride (NH2OH·HCl) in the presence of L-cysteine methyl ester hydrochloride (HSCH2CH(NH2)COOCH3·HCl) as a capping agent. These AuNPs, which achieve higher cell viability in comparison to cetyl trimethyl ammonium bromide (CTAB, a surfactant)-capped counterparts, have demonstrated potential for siRNA delivery. However, it is well known that systemic administration of cationic delivery systems without biological stablising moieties causes non-specific binding with negatively charged serum proteins, resulting in particle aggregation and opsonisation. Consequently, highly stable AuNPs capped with l-cysteine methyl ester hydrochloride conjugated to poly(ethylene glycol) (PEG) were synthesised in this study. PEGylation enhanced the biocompatibility of the AuNPs by reducing toxicity in a range of cell types, by inhibiting interaction with serum proteins thus avoiding aggregation, and, by providing protection against degradation by nucleases. Moreover, these PEGylated AuNPs formed nanoparticles (NPs) with siRNA (which was first compacted with protamine), and had a diameter within the nanoscale range (∼ 250 nm) and a near neutral surface charge (∼ 10 mV). In the future a bifunctional PEG chain on the AuNPs (i.e., SH-PEG-NH2, SH-PEG-COOH) will be used to facilitate conjugation of a targeting ligand to enhance cell specific uptake.

Pegylation increases platelet biocompatibility of gold nanoparticles.

The increasing use of gold nanoparticles in medical diagnosis and treatment has raised the concern over their blood compatibility. The interactions of nanoparticles with blood components may lead to platelet aggregation and endothelial dysfunction. Therefore, medical applications of gold nanoparticles call for increased nanoparticle stability and biocompatibility. Functionalisation of nanoparticles with polythelene glycol (PEGylation) is known to modulate cell-particle interactions. Therefore, the aim of the current study was to investigate the effects of PEGylated-gold nanoparticles on human platelet function and endothelial cells in vitro. Gold nanoparticles, 15 nm in diameter, were synthesised in water using sodium citrate as a reducing and stabilising agent. Functionalised polyethylene glycol-based thiol polymers were used to coat and stabilise pre-synthesised gold nanoparticles. The interaction of gold nanoparticles-citrate and PEGylated-gold nanoparticles with human platelets was measured by Quartz Crystal Microbalance with Dissipation. Platelet-nanoparticles interaction was imaged using phase-contrast, scanning and transmission electron microscopy. The inflammatory effects of gold nanoparticles-citrate and PEGylated-gold nanoparticles in endothelial cells were measured by quantitative real time polymerase chain reaction. PEGylated-gold nanoparticles were stable under physiological conditions and PEGylated-gold nanoparticles-5400 and PEGylated-gold nanoparticles-10800 did not affect platelet aggregation as measured by Quartz Crystal Microbalance with Dissipation. In addition, PEGylated-gold nanoparticles did not induce an inflammatory response when incubated with endothelial cells. Therefore, this study shows that PEGylated-gold nanoparticles with a higher molecular weight of the polymer chain are both platelet- and endothelium-compatible making them attractive candidates for biomedical applications.

The sensitivity of random polymer brush-lamellar polystyrene-b-polymethylmethacrylate block copolymer systems to process conditions.

The use of random copolymer brushes (polystyrene-r-polymethylmethacrylate--PS-r-PMMA) to 'neutralise' substrate surfaces and ordain perpendicular orientation of the microphase separated lamellae in symmetric polystyrene-b-polymethylmethacrylate (PS-b-PMMA) block copolymers (BCPs) is well known. However, less well known is how the brushes interact with both the substrate and the BCP, and how this might change during thermal processing. A detailed study of changes in these films for different brush and diblock PS-b-PMMA molecular weights is reported here. In general, self-assembly and pattern formation is altered little, and a range of brush molecular weights are seen to be effective. However, on extended anneal times, the microphase separated films can undergo dimension changes and loss of order. This process is not related to any complex microphase separation dynamics but rather a degradation of methacrylate components in the film. The data suggest that care must be taken in interpretation of structural changes in these systems as being due to BCP only effects.

The use of quartz crystal microbalance with dissipation (QCM-D) for studying nanoparticle-induced platelet aggregation.

Interactions between blood platelets and nanoparticles have both pharmacological and toxicological significance and may lead to platelet activation and aggregation. Platelet aggregation is usually studied using light aggregometer that neither mimics the conditions found in human microvasculature nor detects microaggregates. A new method for the measurement of platelet microaggregation under flow conditions using a commercially available quartz crystal microbalance with dissipation (QCM-D) has recently been developed. The aim of the current study was to investigate if QCM-D could be used for the measurement of nanoparticle-platelet interactions. Silica, polystyrene, and gold nanoparticles were tested. The interactions were also studied using light aggregometry and flow cytometry, which measured surface abundance of platelet receptors. Platelet activation was imaged using phase contrast and scanning helium ion microscopy. QCM-D was able to measure nanoparticle-induced platelet microaggregation for all nanoparticles tested at concentrations that were undetectable by light aggregometry and flow cytometry. Microaggregates were measured by changes in frequency and dissipation, and the presence of platelets on the sensor surface was confirmed and imaged by phase contrast and scanning helium ion microscopy.