In vitro coronal protein signatures and biological impact of silver nanoparticles synthesized with different natural polymers as capping agents.

Biopolymer-capped particles, sodium alginate-, gelatin- and reconstituted silk fibroin-capped nanosilver (AgNPs), were synthesized with an intention to study, simultaneously, their in vitro and in vivo haemocompatibility, one of the major safety factors in biomedical applications. Solid state characterization showed formation of spherical nanoparticles with 5 to 30 nm primary sizes (transmission electron microscopy) and X-ray photoelectron spectroscopy analysis of particles confirmed silver bonding with the biopolymer moieties. The degree of aggregation of the biopolymer-capped AgNPs in the synthesis medium (ultrapure water) is relatively low, with comparable hydrodynamic size with those of the control citrate-stabilized NPs, and remained relatively unchanged even after 6 weeks. The polymer-capped nanoparticles showed different degrees of aggregation in biologically relevant media - PBS (pH 7.4) and 2% human blood plasma - with citrate- (control) and alginate-capped particles showing the highest aggregation, while gelatin- and silk fibroin-capped particles revealed better stability and less aggregation in these media. In vitro cytotoxicity studies revealed that the polymer-capped particles exhibited both concentration and (hydrodynamic) size-dependent haemolytic activity, the extent of which was higher (up to 100% in some cases) in collected whole blood samples of healthy human volunteers when compared to that in the washed erythrocytes. This difference is thought to result from the detected protein corona formation on the nanoparticle surface in the whole blood system, which was associated with reduced particle aggregation, causing more severe cytotoxic effects. At the tested particle concentration range in vitro, we observed a negligible haemolysis effect in vivo (Balb/c mice). Polymer-capped particles did accumulate in organs, with the highest levels detected in the liver (up to 422 µg per g tissue), yet no adverse behavioural effects were observed in the mice during the duration of the nanoparticle exposure.

Understanding the performance of a paper-based UV exposure sensor: The photodegradation mechanism of brilliant blue FCF in the presence of TiO2 photocatalysts in both the solid state and solution.

RATIONALE: The decolouration of brilliant blue FCF by the action of titanium dioxide (TiO2 ) under ultraviolet (UV) exposure has been recently reported as the basis of a paper-based sensor for monitoring UV sun exposure. The mechanism of brilliant blue FCF photodegradation in the presence of the photocatalyst and the resulting photoproducts are thus far unknown. METHODS: The UV-initiated photodegradation of brilliant blue FCF in the presence of TiO2 for both the aqueous and the solid state was investigated. Degradation in the solid state was observed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-MS). Decomposition of the dye in the aqueous state was analyzed using liquid chromatography/mass spectrometry (LC/MS) and ultraviolet-visible (UV-Vis) spectroscopy. RESULTS: After UV radiation exposure, the brilliant blue FCF base peak [M1 + NH4 ]+ (m/z calc. 766.194 found 766.194) decreased in the LC/MS chromatogram with a concomitant appearance of BB-FCF decomposition products involving the sequential loss of the N-ethyl and N-methylbenzene sulfonate (MBSA) groups, assigned as [M2 + H]+ (-MBSA, calc. 579.163 found 579.162), [M3 + H]+ (-MBSA, -Et, calc. 551.131 found 551.131), [M4 + H]+ (-2MBSA, calc. 409.158 found 409.158), [M5 + H]+ (-2MBSA, -Et, calc. 381.127 found 381.127). Ions [M2 + H]+ and [M3 + H]+ were also identified in the photodegradation products using MALDI-MS. Observation by UV-Vis indicated a decrease in the solution absorbance maxima and an associated blue-shift upon UV exposure in solution. CONCLUSIONS: The LC/MS analysis indicated two main oxidation processes. The most obvious was attack of the N-methylene, eliminating either ethyl or MBSA groups. The presence of the hydroxylated decomposition product M13 ([M13 + H]+ , calc. 595.157 found 595.157) supported this assignment. In addition, the detection of photoproduct M8, proposed to be 3-((ethylamino)methyl)benzenesulfonic acid ([M8 + H]+ , calc. 216.069 found 216.069), indicates an aryl-oxidative elimination. The absence of the aryl-hydroxy products normally expected to accompany the formation of M8 is proposed to be due to TiO2 -binding catechol-like derivatives, which are then removed upon filtration.

Micropatterning of porous silicon Bragg reflectors with poly(ethylene glycol) to fabricate cell microarrays: Towards single cell sensing.

The work presented here describes the development of an optical label-free biosensor based on a porous silicon (PSi) Bragg reflector to study heterogeneity in single cells. Photolithographic patterning of a poly(ethylene glycol) (PEG) hydrogel with a photoinitiator was employed on RGD peptide-modified PSi to create micropatterns with cell adhesive and cell repellent areas. Macrophage J774 cells were incubated to form cell microarrays and single cell arrays. Moreover, cells on the microarrays were lysed osmotically with Milli-Q™ water and the infiltration of cell lysate into the porous matrix was monitored by measuring the red shift in the reflectivity. On average, the magnitude of red shift increased with the increase in the number of cells on the micropatterns. The red shift from the spots with single cells varied from spot to spot emphasizing the heterogeneous nature of the individual cells.

Reversible Thermoresponsive Plasmonic Core-Satellite Nanostructures That Exhibit Both Expansion and Contraction (UCST and LCST).

The assembly of sophisticated gold nanoparticle constructs where thermoresponsive core-satellite nanostructures are created and the satellites are close enough to the core for strong surface plasmon resonance coupling to occur, has begun to be developed. The linker between the core and the satellites being a thermoresponsive polymer means that a dispersion of these nanostructures will show temperature-dependent optical properties as the distance between the core and the satellites changes. Unlike previous related thermoresponsive core-satellite systems that undergo a single thermoresponsive transition, herein a polymer system with dual thermoresponsive transitions (block copolymer with both lower critical solution temperature and upper critical solution temperature) is employed as a linker that modulates the gap distance between the "core" and "satellites" in response to the temperature. In this way, optical properties of dispersions can be dynamically tuned. The system permits wide and reversible control of the optical properties, which may render them excellent candidates for reversible nanosensors.

Thermoresponsive plasmonic core-satellite nanostructures with reversible, temperature sensitive optical properties.

Herein is described a facile method for the assembly of plasmonic gold nanoparticles into smart plasmonic core-satellite nanostructures that allow for the dynamic and reversible tuning of the localised surface plasmon resonance using temperature. This smart system takes advantage of the thermoresponsive polymer linker that modulates the gap distance between the core and satellites in response to the temperature, resulting in the tuning of the surface plasmon coupling and resultant optical shift. It permits optical shifts over a wide wavelength range and reversible control of the optical properties by altering the temperature, which may allow these systems to become candidates for temperature sensitive nanosensors.

Can single molecule localization microscopy be used to map closely spaced RGD nanodomains?

Cells sense and respond to nanoscale variations in the distribution of ligands to adhesion receptors. This makes single molecule localization microscopy (SMLM) an attractive tool to map the distribution of ligands on nanopatterned surfaces. We explore the use of SMLM spatial cluster analysis to detect nanodomains of the cell adhesion-stimulating tripeptide arginine-glycine-aspartic acid (RGD). These domains were formed by the phase separation of block copolymers with controllable spacing on the scale of tens of nanometers. We first determined the topology of the block copolymer with atomic force microscopy (AFM) and then imaged the localization of individual RGD peptides with direct stochastic optical reconstruction microscopy (dSTORM). To compare the data, we analyzed the dSTORM data with DBSCAN (density-based spatial clustering application with noise). The ligand distribution and polymer topology are not necessary identical since peptides may attach to the polymer outside the nanodomains and/or coupling and detection of peptides within the nanodomains is incomplete. We therefore performed simulations to explore the extent to which nanodomains could be mapped with dSTORM. We found that successful detection of nanodomains by dSTORM was influenced by the inter-domain spacing and the localization precision of individual fluorophores, and less by non-specific absorption of ligands to the substratum. For example, under our imaging conditions, DBSCAN identification of nanodomains spaced further than 50 nm apart was largely independent of background localisations, while nanodomains spaced closer than 50 nm required a localization precision of ~11 nm to correctly estimate the modal nearest neighbor distance (NDD) between nanodomains. We therefore conclude that SMLM is a promising technique to directly map the distribution and nanoscale organization of ligands and would benefit from an improved localization precision.

A versatile method for the preparation of carbon-rhodium hybrid catalysts on graphene and carbon black.

Strategies for combining the selectivity and efficiency of homogeneous organometallic catalysts with the versatility of heterogeneous catalysts are urgently needed. Herein a direct and modular methodology is presented that provides rapid access to well-defined carbon-rhodium hybrid catalysts. A pre-synthesized Rh(i) complex containing a carbene-triazole ligand was found to be stable for direct immobilization onto unactivated graphene, carbon black and glassy carbon electrodes. Characterization of the heterogeneous systems using X-ray photoelectron spectroscopy (XPS), thermogravimetric analysis (TGA), inductively coupled plasma-optical emission spectroscopy/mass spectrometry (ICP-OES/MS), Raman spectroscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) confirmed the well-defined nature of the hybrid catalysts. The hybrid catalysts show excellent activity, comparable to that of the homogeneous system for the hydrosilylation of diphenylacetylene, with turnover numbers ranging from 5000 to 48 000. These catalysts are the best reported to date for the hydrosilylation of diphenylacetylene. In common with conventional heterogeneous catalysts, high reusability, due to a lack of Rh metal leaching, was also observed for all carbon-rhodium complexes under investigation.

Simple Method for Tuning the Optical Properties of Thermoresponsive Plasmonic Nanogels.

We report a straightforward way for forming and tuning the optical properties of thermally responsive plasmonic nanogels. Upon functionalization, a small red shift (2-3 nm) of the pNIPAM@AuNPs was observed due to changes in the refractive index surrounding the AuNP. By adding thermoresponsive poly-N-isopropylacrylamide (pNIPAM) into the pNIPAM@AuNP, its optical response was significantly increased. Heating the nanogel such that the pNIPAM collapsed and acted as a cross-link resulted in the aggregation of the AuNPs. The plasmonic response with red shifts of up to 20 nm was observed. The enlarged red shift was due to the increase in the dielectric constant around the particles and the interparticle interaction of the AuNPs. The interparticle interaction also leads to the broadening of the spectra. Experimental data and finite-difference time-domain (FDTD) calculation are in agreement with this observation. The temperature-dependent optical properties were reversible through multiple cycles of heating and cooling.

Reversible gating of smart plasmonic molecular traps using thermoresponsive polymers for single-molecule detection.

Single-molecule surface-enhanced Raman spectroscopy (SERS) has attracted increasing interest for chemical and biochemical sensing. Many conventional substrates have a broad distribution of SERS enhancements, which compromise reproducibility and result in slow response times for single-molecule detection. Here we report a smart plasmonic sensor that can reversibly trap a single molecule at hotspots for rapid single-molecule detection. The sensor was fabricated through electrostatic self-assembly of gold nanoparticles onto a gold/silica-coated silicon substrate, producing a high yield of uniformly distributed hotspots on the surface. The hotspots were isolated with a monolayer of a thermoresponsive polymer (poly(N-isopropylacrylamide)), which act as gates for molecular trapping at the hotspots. The sensor shows not only a good SERS reproducibility but also a capability to repetitively trap and release molecules for single-molecular sensing. The single-molecule sensitivity is experimentally verified using SERS spectral blinking and bianalyte methods.

Heat-treated stainless steel felt as scalable anode material for bioelectrochemical systems.

This work reports a simple and scalable method to convert stainless steel (SS) felt into an effective anode for bioelectrochemical systems (BESs) by means of heat treatment. X-ray photoelectron spectroscopy and cyclic voltammetry elucidated that the heat treatment generated an iron oxide rich layer on the SS felt surface. The iron oxide layer dramatically enhanced the electroactive biofilm formation on SS felt surface in BESs. Consequently, the sustained current densities achieved on the treated electrodes (1 cm(2)) were around 1.5±0.13 mA/cm(2), which was seven times higher than the untreated electrodes (0.22±0.04 mA/cm(2)). To test the scalability of this material, the heat-treated SS felt was scaled up to 150 cm(2) and similar current density (1.5 mA/cm(2)) was achieved on the larger electrode. The low cost, straightforwardness of the treatment, high conductivity and high bioelectrocatalytic performance make heat-treated SS felt a scalable anodic material for BESs.

Polymersomes prepared from thermoresponsive fluorescent protein-polymer bioconjugates: capture of and report on drug and protein payloads.

Polymersomes provide a good platform for targeted drug delivery and the creation of complex (bio)catalytically active systems for research in synthetic biology. To realize these applications requires both spatial control over the encapsulation components in these polymersomes and a means to report where the components are in the polymersomes. To address these twin challenges, we synthesized the protein-polymer bioconjugate PNIPAM-b-amilFP497 composed of thermoresponsive poly(N-isopropylacrylamide) (PNIPAM) and a green-fluorescent protein variant (amilFP497). Above 37 °C, this bioconjugate forms polymersomes that can (co-)encapsulate the fluorescent drug doxorubicin and the fluorescent light-harvesting protein phycoerythrin 545 (PE545). Using fluorescence lifetime imaging microscopy and Förster resonance energy transfer (FLIM-FRET), we can distinguish the co-encapsulated PE545 protein inside the polymersome membrane while doxorubicin is found both in the polymersome core and membrane.

Synthesis and high-throughput processing of polymeric hydrogels for 3D cell culture.

3D cell cultures have drawn a large amount of interest in the scientific community with their ability to closely mimic physiological conditions. Hydrogels have been used extensively in the development of extracellular matrix (ECM) mimics for 3D cell culture. Compounds such as collagen and fibrin are commonly used to synthesize natural ECM mimics; however they suffer from batch-to-batch variation. In this Review we explore the synthesis route of hydrogels; how they can be altered to give different chemical and physical properties; how different biomolecules such as arginylglycylaspartic acid (RGD) or vascular endothelial growth factor (VEGF) can be incorporated to give different biological cues; and how to create concentration gradients with UV light. There will also be emphasis on the types of techniques available in high-throughput processing such as nozzle and droplet-based biofabrication, photoenabled biofabrication, and microfluidics. The combination of these approaches and techniques allow the preparation of hydrogels which are capable of mimicking the ECM.

Chemical patterning on preformed porous silicon photonic crystals: towards multiplex detection of protease activity at precise positions†Electronic supplementary information (ESI) available: SEM images, XPS result and more optical reflectivity data. See DOI: 10.1039/c4tb00281dClick here for additional data file.

Porous silicon (PSi) rugate filters modified with alkyne-terminated monolayers were chemically patterned using a combination of photolithography of photoresist and click chemistry. Two chemical functionalities were obtained by conjugating, via click reactions, ethylene glycol moieties containing two different terminal groups to discrete areas towards the exterior of a PSi rugate filter. The patterning of biological species to the functionalized surface was demonstrated through the conjugation of fluorescein isothiocyanate labelled bovine serum albumin (FITC-BSA). Fluorescence microscopy showed selective positioning of FITC-BSA at discretely functionalized areas. Meanwhile, the optical information from precisely defined positions on the patterned surface was monitored by optical reflectivity measurements. The optical measurements revealed successful step-wise chemical functionalization followed by immobilization of gelatin. Multiplex detection of protease activity from different array elements on the patterned surface was demonstrated by monitoring the blue shifts in the reflectivity spectra resulted from the digestion of gelatin by subtilisin. Precise information from both individual elements and average population was acquired. This technique is important for the development of PSi into a microarray platform for highly parallel biosensing applications, especially for cell-based assays.

Flame oxidation of stainless steel felt enhances anodic biofilm formation and current output in bioelectrochemical systems.

Stainless steel (SS) can be an attractive material to create large electrodes for microbial bioelectrochemical systems (BESs), due to its low cost and high conductivity. However, poor biocompatibility limits its successful application today. Here we report a simple and effective method to make SS electrodes biocompatible by means of flame oxidation. Physicochemical characterization of electrode surface indicated that iron oxide nanoparticles (IONPs) were generated in situ on an SS felt surface by flame oxidation. IONPs-coating dramatically enhanced the biocompatibility of SS felt and consequently resulted in a robust electroactive biofilm formation at its surface in BESs. The maximum current densities reached at IONPs-coated SS felt electrodes were 16.5 times and 4.8 times higher than the untreated SS felts and carbon felts, respectively. Furthermore, the maximum current density achieved with the IONPs-coated SS felt (1.92 mA/cm(2), 27.42 mA/cm(3)) is one of the highest current densities reported thus far. These results demonstrate for the first time that flame oxidized SS felts could be a good alternative to carbon-based electrodes for achieving high current densities in BESs. Most importantly, high conductivity, excellent mechanical strength, strong chemical stability, large specific surface area, and comparatively low cost of flame oxidized SS felts offer exciting opportunities for scaling-up of the anodes for BESs.

Light-induced organic monolayer modification of iodinated carbon electrodes.

We report the modification of carbon electrodes formed from pyrolyzed photoresist films (PPF) via plasma iodination followed by the organic monolayer modification of these surfaces. The iodinated surfaces were characterized using cyclic voltammetry, atomic force microscopy, and X-ray photoelectron spectroscopy to enable the optimization of the iodination while preserving the stability and smoothness of the carbon surface. Subsequently, the C-I surface was further modified with molecules that possess an alkene or alkyne at one end through light activation with low energy (visible range λ 514 nm). The versatility of the modification reaction of the C-I surfaces is shown by reactions with undecylenic acid, 1,8-nonadiyne, and S-undec-10-enyl-2,2,2-trifluoroethanethioate (C11-S-TFA). Modification with 1,8-nonadiyne allows further modification via "click" chemistry with azido-terminated oligo(ethylene oxide) molecules demonstrated briefly to alter the hydrophilicity of the surface after attachment of ethylene oxide moieties. Furthermore, patterning of C11-S-TFA was demonstrated using a simple photolithography technique. Deprotection of the C11-S-TFA gave a free thiol allowed patterning of gold nanoparticles on the surface as verified using scanning electron microscopy (SEM). These results demonstrate that plasma iodination to form C-I is a versatile, simple, and modular approach to functionalize the carbon surface.

Stimuli-responsive functionalized mesoporous silica nanoparticles for drug release in response to various biological stimuli.

A silica-based mesoporous nanosphere (MSN) controlled-release drug delivery system has been synthesized and characterized. The system uses l-cysteine derivatized gold nanoparticles (AuNPs), bound to the MSNs using Cu2+ as a bridging ion. The AuNPs serve as removable caps that hinder the release of drug molecules inside the amino functionalized MSN mesoporous framework. The modified MSNs themselves exhibit negligible cytotoxicity to living cells, as revealed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The drug delivery system requires one of two biological stimuli to trigger drug release. These stimuli are either: low pH (pH < 5); or elevated levels of adenosine triphosphate (ATP) (concentration > 4 mM). The feasibility of biologically controlled release was demonstrated through the stimuli-induced removal of the AuNP caps over the MSN releasing the anticancer drug doxorubicin. We envisage that this MSN system could play a significant role in developing new generations of controlled-release delivery vehicles.

High-order multiblock copolymers via iterative Cu(0)-mediated radical polymerizations (SET-LRP): toward biological precision.

We report a new approach for the facile synthesis of high-order multiblock copolymers comprising very short blocks. The approach entails sequential addition of different monomers via an iterative single electron transfer-living radical polymerization technique, allowing nearly perfect control of the copolymer microstructure. It is possible to synthesize high-order multiblock copolymers with unprecedented control, i.e., A-B-C-D-E-etc., without any need for purification between iterative 24 h block formation steps. To illustrate this concept, we report the synthesis of model P(MA-b-MA...) homopolymer and P(MA-b-nBuA-b-EA-b-2EHA-b-EA-b-nBuA) copolymer in extremely high yield. Finally, the halide end-group can be modified via "click chemistry", including thiol-bromide click chemistry, sodium methanethiosulfonate nucleophilic substitution, and atom transfer radical nitroxide coupling reaction, to yield functional, structurally complex macromolecules.

Post-functionalization of ATRP polymers using both thiol/ene and thiol/disulfide exchange chemistry.

In this communication, we report on a new route to the functionalization of ATRP polymers exploiting their halide end-groups, which were converted successfully into reactive disulfide end-groups, using sodium methanethiosulfonate. The resultant disulfide-terminated polymers could then be reacted with different functional thiols to yield functional polymers exploiting either thiol/disulfide exchange chemistry or thiol/ene "click" reactions.

High fidelity vinyl terminated polymers by combining RAFT and cobalt catalytic chain transfer (CCT) polymerization methods.

In this work, we combine reversible addition fragmentation transfer polymerization (RAFT) with cobalt catalytic chain transfer (CCT) to obtain omega-unsaturated polymers with relatively low polydispersity values.