Liquid Crystal Nanoparticle Conjugates for Scavenging Reactive Oxygen Species in Live Cells.

The elevated intracellular production of or extracellular exposure to reactive oxygen species (ROS) causes oxidative stress to cells, resulting in deleterious irreversible biomolecular reactions (e.g., lipid peroxidation) and disease progression. The use of low-molecular weight antioxidants, such as 4-amino-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO), as ROS scavengers fails to achieve the desired efficacy because of their poor or uncontrolled cellular uptake and off-target effects, such as dysfunction of essential redox homeostasis. In this study, we fabricated a liquid crystal nanoparticle (LCNP) conjugate system with the fluorescent dye perylene (PY) loaded in the interior and poly (ethylene glycol) (PEG) decorated on the surface along with multiple molecules of TEMPO (PY-LCNP-PEG/TEMPO). PY-LCNP-PEG/TEMPO exhibit enhanced cellular uptake, and efficient ROS-scavenging activity in live cells. On average, the 120 nm diameter PY-LCNPs were conjugated with >1800 molecules of TEMPO moieties on their surface. PY-LCNP-PEG/TEMPO showed significantly greater reduction in ROS activity and lipid peroxidation compared to free TEMPO when the cells were challenged with ROS generating agents, such as hydrogen peroxide (H2O2). We suggest that this is due to the increased local concentration of TEMPO molecules on the surface of the PY-LCNP-PEG/TEMPO NPs, which efficiently bind to the plasma membrane and enter cells. Overall, these results demonstrate the enhanced capability of TEMPO-conjugated LCNPs to protect live cells from oxidative stress by effectively scavenging ROS and reducing lipid peroxidation.

Light tunable plasmonic metasurfaces.

Self-assembled plasmonic metasurfaces are promising optical platforms to achieve accessible flat optics, due to their strong light-matter interaction, nanometer length scale precision, large area, light weight, and high-throughput fabrication. Here, using photothermal continuous wave laser lithography, we show the spectral and spatial tuning of metasurfaces comprised of a monolayer of ligand capped hexagonally packed gold nanospheres. To tune the spectral response of the metasurfaces, we show that by controlling the intensity of a laser focused onto the metasurface that the absorption peak can be reconfigured from the visible to near-infrared wavelength. The irreversible spectral tuning mechanism is attributed to photothermal modification of the surface morphology. Combining self-assembled metasurfaces with laser lithography, we demonstrate an optically thin (λ/42), spectrally selective plasmonic Fresnel zone plate. This work establishes a new pathway for creating flat, large area, frequency selective optical elements using self-assembled plasmonic metasurfaces and laser lithography.

Analyzing fidelity and reproducibility of DNA templated plasmonic nanostructures.

Synthetic DNA templated nanostructures offer an excellent platform for the precise spatial and orientational positioning of organic and inorganic nanomaterials. Previous reports have shown its applicability in the organization of plasmonic nanoparticles in a number of geometries for the purpose of realizing tunable nanoscale optical devices. However, translation of nanoparticle-DNA constructs to application requires additional efforts to increase scalability, reproducibility, and formation yields. Understanding all these factors is, in turn, predicated on in-depth analysis of each structure and comparing how formation changes with complexity. Towards the latter goal, we assemble seven unique plasmonic nanostructure symmetries of increasing complexity based on assembly of gold nanorods and nanoparticles on two different DNA origami templates, a DNA triangle and rhombus, and characterize them using gel electrophoresis, atomic force- and transmission electron microscopy, as well as optical spectroscopy. In particular, we focus on how much control can be elicited over yield, reproducibility, shape, size, inter-particle angles, gaps, and plasmon shifts as compared to expectations from computer simulations as structural complexity increases. We discuss how these results can contribute to establishing process principles for creating DNA templated plasmonic nanostructures.

Dynamic Plasmonic Pixels.

Information display utilizing plasmonic color generation has recently emerged as an alternative paradigm to traditional printing and display technologies. However, many implementations so far have either presented static pixels with a single display state or rely on relatively slow switching mechanisms such as chemical transformations or liquid crystal transitions. Here, we demonstrate spatial, spectral, and temporal control of light using dynamic plasmonic pixels that function through the electric-field-induced alignment of plasmonic nanorods in organic suspensions. By tailoring the geometry and composition (Au and Au@Ag) of the nanorods, we illustrate light modulation across a significant portion of the visible and infrared spectrum (600-2400 nm). The fast (∼30 µs), reversible nanorod alignment is manifested as distinct color changes, characterized by shifts of observed chromaticity and luminance. Integration into larger device architectures is showcased by the fabrication of a seven-segment numerical indicator. The control of light on demand achieved in these dynamic plasmonic pixels establishes a favorable platform for engineering high-performance optical devices.

Hybrid Liquid Crystal Nanocarriers for Enhanced Zinc Phthalocyanine-Mediated Photodynamic Therapy.

Current challenges in photodynamic therapy (PDT) include both the targeted delivery of the photosensitizer (PS) to the desired cellular location and the maintenance of PS efficacy. Zinc phthalocyanine (ZnPc), a macrocyclic porphyrin and a potent PS for PDT, undergoes photoexcitation to generate reactive singlet oxygen that kills cells efficiently, particularly when delivered to the plasma membrane. Like other commonly employed PS, ZnPc is highly hydrophobic and prone to self-aggregation in aqueous biological media. Further, it lacks innate subcellular targeting specificity. Cumulatively, these attributes pose significant challenges for delivery via traditional systemic drug delivery modalities. Here, we report the development and characterization of a liquid crystal nanoparticle (LCNP)-based formulation for the encapsulation and targeted tethering of ZnPc to the plasma membrane bilayer. ZnPc was coloaded with the organic fluorophore, perylene (PY), in the hydrophobic polymeric matrix of the LCNP core. PY facilitated the fluorescence-based tracking of the LCNP carrier while also serving as a Förster resonance energy transfer (FRET) donor to the ZnPc acceptor. This configuration availed efficient singlet oxygen generation via enhanced excitation of ZnPc from multiple surrounding PY energy donors. When excited in a FRET configuration, cuvette-based assays revealed that singlet oxygen generation from the ZnPc was ∼1.8-fold greater and kinetically 12 times faster compared to when the ZnPc was excited directly. The specific tethering of the LCNPs to the plasma membrane of HEK 293 T/17 and HeLa cells was achieved by surface functionalization of the NPs with PEGylated cholesterol. In HeLa cells, LCNPs coloaded with PY and ZnPc, when photoexcited in a FRET configuration, mediated 70% greater cell killing compared to LCNPs containing ZnPc alone (direct excitation of ZnPc). This was attributed to a significant increase of the oxidative stress in the cells during the PDT. Overall, this work details the ability of the LCNP platform to facilitate (1) the specific tethering of the PY-ZnPc FRET pair to the plasma membrane and (2) the FRET-mediated, augmented singlet oxygen generation for enhanced PDT relative to the direct excitation of ZnPc alone.

Linear and nonlinear optical characterization of self-assembled, large-area gold nanosphere metasurfaces with sub-nanometer gaps: errata.

We correct a nomenclature error for the plasmon ruler equation used to fit the simulation data in Fig. 2(d) [Opt. Express24, 27360 (2016)].

Targeted Plasma Membrane Delivery of a Hydrophobic Cargo Encapsulated in a Liquid Crystal Nanoparticle Carrier.

The controlled delivery of drug/imaging agents to cells is critical for the development of therapeutics and for the study of cellular signaling processes. Recently, nanoparticles (NPs) have shown significant promise in the development of such delivery systems. Here, a liquid crystal NP (LCNP)-based delivery system has been employed for the controlled delivery of a water-insoluble dye, 3,3'-dioctadecyloxacarbocyanine perchlorate (DiO), from within the NP core to the hydrophobic region of a plasma membrane bilayer. During the synthesis of the NPs, the dye was efficiently incorporated into the hydrophobic LCNP core, as confirmed by multiple spectroscopic analyses. Conjugation of a PEGylated cholesterol derivative to the NP surface (DiO-LCNP-PEG-Chol) enabled the binding of the dye-loaded NPs to the plasma membrane in HEK 293T/17 cells. Time-resolved laser scanning confocal microscopy and Förster resonance energy transfer (FRET) imaging confirmed the passive efflux of DiO from the LCNP core and its insertion into the plasma membrane bilayer. Finally, the delivery of DiO as a LCNP-PEG-Chol attenuated the cytotoxicity of DiO; the NP form of DiO exhibited ~30-40% less toxicity compared to DiOfree delivered from bulk solution. This approach demonstrates the utility of the LCNP platform as an efficient modality for the membrane-specific delivery and modulation of hydrophobic molecular cargos.

Linear and nonlinear optical characterization of self-assembled, large-area gold nanosphere metasurfaces with sub-nanometer gaps.

We created centimeter-scale area metasurfaces consisting of a quasi-hexagonally close packed monolayer of gold nanospheres capped with alkanethiol ligands on glass substrates using a directed self-assembly approach. We experimentally characterized the morphology and the linear and nonlinear optical properties of metasurfaces. We show these metasurfaces, with interparticle gaps of 0.6 nm, are modeled well using a classical (without charge transfer) description. We find a large dispersion of linear refractive index, ranging from values less than vacuum, 0.87 at 600 nm, to Germanium-like values of 4.1 at 880 nm, determined using spectroscopic ellipsometry. Nonlinear optical characterization was carried out using femtosecond Z-scan and we observe saturation behavior of the nonlinear absorption (NLA) and nonlinear refraction (NLR). We find a negative NLR from these metasurfaces two orders of magnitude larger (n2,sat = -7.94x10-9 cm2/W at Isat,n2 = 0.43 GW/cm2) than previous reports on gold nanostructures at similar femtosecond time scales. We also find the magnitude of the NLA comparable to the largest values reported (ß2,sat = -0.90x105 cm/GW at Isat,ß2 = 0.34 GW/cm2). Precise knowledge of the index of refraction is of crucial importance for emerging dispersion engineering technologies. Furthermore, utilizing this directed self-assembly approach enables the nanometer scale resolution required to develop the unique optical response and simultaneously provides high-throughput for potential device realization.

Electrically Induced Twist in Smectic Liquid-Crystalline Elastomers.

As an approach for electrically controllable actuators, we prepare elastomers of chiral smectic-A liquid crystals, which have an electroclinic effect, i.e., molecular tilt induced by an applied electric field. Surprisingly, our experiments find that an electric field causes a rapid and reversible twisting of the film out of the plane, with a helical sense that depends on the sign of the field. To explain this twist, we develop a continuum elastic theory based on an asymmetry between the front and back of the film. We further present finite-element simulations, which show the dynamic shape change.

Lipid Raft-Mediated Membrane Tethering and Delivery of Hydrophobic Cargos from Liquid Crystal-Based Nanocarriers.

A main goal of bionanotechnology and nanoparticle (NP)-mediated drug delivery (NMDD) continues to be the development of novel biomaterials that can controllably modulate the activity of the NP-associated therapeutic cargo. One of the desired subcellular locations for targeted delivery in NMDD is the plasma membrane. However, the controlled delivery of hydrophobic cargos to the membrane bilayer poses significant challenges including cargo precipitation and lack of specificity. Here, we employ a liquid crystal NP (LCNP)-based delivery system for the controlled partitioning of a model dye cargo from within the NP core into the plasma membrane bilayer. During synthesis of the NPs, the water-insoluble model dye cargo, 3,3'-dioctadecyloxacarbocyanine perchlorate (DiO), was efficiently incorporated into the hydrophobic LCNP core as confirmed by multiple spectroscopic analyses. Conjugation of a PEGylated cholesterol derivative to the NP surface (DiO-LCNP-PEG-Chol) facilitated the localization of the dye-loaded NPs to lipid raft microdomains in the plasma membrane in HEK 293T/17 cell. Analysis of DiO cellular internalization kinetics revealed that when delivered as a LCNP-PEG-Chol NP, the half-life of DiO membrane residence time (30 min) was twice that of free DiO (DiO(free)) (15 min) delivered from bulk solution. Time-resolved laser scanning confocal microscopy was employed to visualize the passive efflux of DiO from the LCNP core and its insertion into the plasma membrane bilayer as confirmed by Förster resonance energy transfer (FRET) imaging. Finally, the delivery of DiO as a LCNP-PEG-Chol complex resulted in the attenuation of its cytotoxicity; the NP form of DiO exhibited ∼30-40% less toxicity compared to DiO(free). Our data demonstrate the utility of the LCNP platform as an efficient vehicle for the combined membrane-targeted delivery and physicochemical modulation of molecular cargos using lipid raft-mediated tethering.

Multifunctional liquid crystal nanoparticles for intracellular fluorescent imaging and drug delivery.

A continuing goal of nanoparticle (NP)-mediated drug delivery (NMDD) is the simultaneous improvement of drug efficacy coupled with tracking of the intracellular fate of the nanoparticle delivery vehicle and its drug cargo. Here, we present a robust multifunctional liquid crystal NP (LCNP)-based delivery system that affords facile intracellular fate tracking coupled with the efficient delivery and modulation of the anticancer therapeutic doxorubicin (Dox), employed here as a model drug cargo. The LCNPs consist of (1) a liquid crystal cross-linking agent, (2) a homologue of the organic chromophore perylene, and (3) a polymerizable surfactant containing a carboxylate headgroup. The NP core provides an environment to both incorporate fluorescent dye for spectrally tuned particle tracking and encapsulation of amphiphilic and/or hydrophobic agents for intracellular delivery. The carboxylate head groups enable conjugation to biologicals to facilitate the cellular uptake of the particles. Upon functionalization of the NPs with transferrin, we show the ability to differentially label the recycling endocytic pathway in HEK 293T/17 cells in a time-resolved manner with minimal cytotoxicity and with superior dye photostability compared to traditional organic fluorophores. Further, when passively loaded with Dox, the NPs mediate the rapid uptake and subsequent sustained release of Dox from within endocytic vesicles. We demonstrate the ability of the LCNPs to simultaneously serve as both an efficient delivery vehicle for Dox as well as a modulator of the drug's cytotoxicity. Specifically, the delivery of Dox as a LCNP conjugate results in a ∼40-fold improvement in its IC50 compared to free Dox in solution. Cumulatively, our results demonstrate the utility of the LCNPs as an effective nanomaterial for simultaneous cellular imaging, tracking, and delivery of drug cargos.

A technique to functionalize and self-assemble macroscopic nanoparticle-ligand monolayer films onto template-free substrates.

This protocol describes a self-assembly technique to create macroscopic monolayer films composed of ligand-coated nanoparticles. The simple, robust and scalable technique efficiently functionalizes metallic nanoparticles with thiol-ligands in a miscible water/organic solvent mixture allowing for rapid grafting of thiol groups onto the gold nanoparticle surface. The hydrophobic ligands on the nanoparticles then quickly phase separate the nanoparticles from the aqueous based suspension and confine them to the air-fluid interface. This drives the ligand-capped nanoparticles to form monolayer domains at the air-fluid interface. The use of water-miscible organic solvents is important as it enables the transport of the nanoparticles from the interface onto template-free substrates. The flow is mediated by a surface tension gradient and creates macroscopic, high-density, monolayer nanoparticle-ligand films. This self-assembly technique may be generalized to include the use of particles of different compositions, size, and shape and may lead to an efficient assembly method to produce low-cost, macroscopic, high-density, monolayer nanoparticle films for wide-spread applications.

Interpenetrating networks based on gelatin methacrylamide and PEG formed using concurrent thiol click chemistries for hydrogel tissue engineering scaffolds.

The integration of biological extracellular matrix (ECM) components and synthetic materials is a promising pathway to fabricate the next generation of hydrogel-based tissue scaffolds that more accurately emulate the microscale heterogeneity of natural ECM. We report the development of a bio/synthetic interpenetrating network (BioSINx), containing gelatin methacrylamide (GelMA) polymerized within a poly(ethylene glycol) (PEG) framework to form a mechanically robust network capable of supporting both internal cell encapsulation and surface cell adherence. The covalently crosslinked PEG network was formed by thiol-yne coupling, while the bioactive GelMA was integrated using a concurrent thiol-ene coupling reaction. The physical properties (i.e. swelling, modulus) of BioSINx were compared to both PEG networks with physically-incorporated gelatin (BioSINP) and homogenous hydrogels. BioSINx displayed superior physical properties and significantly lower gelatin dissolution. These benefits led to enhanced cytocompatibility for both cell adhesion and encapsulation; furthermore, the increased physical strength provided for the generation of a micro-engineered tissue scaffold. Endothelial cells showed extensive cytoplasmic spreading and the formation of cellular adhesion sites when cultured onto BioSINx; moreover, both encapsulated and adherent cells showed sustained viability and proliferation.

Hydrodynamic shaping, polymerization, and subsequent modification of thiol click fibers.

Hydrodynamic focusing in microfluidic channels is used to produce highly uniform, shaped polymer fibers at room temperature and under "green" conditions. Core streams of thiol-ene and thiol-yne prepolymer solutions were guided using a phase-matched sheath stream through microfluidic channels with grooved walls to determine shape. Size was dictated by the ratio of the flow rates of the core and sheath streams. Thiol click reactions were initiated using UV illumination to lock in predesigned cross-sectional shapes and sizes. This approach proved to be much more flexible than electrospinning in that highly uniform fibers can be produced from prepolymer solutions with varying compositions and viscosities with made-to-order sizes and shapes. Furthermore, a very simple manipulation of the composition provided reactive groups on the fiber surface for attachment of active ligands and biological components. A proof-of-principle experiment demonstrated that biotin attached to thiol groups on the fiber surface could specifically bind a fluorescent protein.

Tuning mechanical properties of liquid crystalline nanoparticles.

We report the synthesis of colloidal nanoparticles with an internal structure forming a gel-like matrix. These nanoparticles are composed of low molecular weight liquid crystal (LC) 4-pentyl-4-cyanobiphenyl (5CB) encapsulated in an LC-based polymer network. Using nanoscopic mechanical analysis, we demonstrate the ability to independently tune the shape anisotropy and stiffness by varying, respectively, the 5CB concentration and the extent of the polymer cross-linking. Based on these data, a model is introduced to account for the effect of the polymer network on the mechanical properties, thus providing novel insight into the nanomechanics of these soft particles.

High-pressure investigations of a ferroelectric liquid crystal exhibiting a trend reversal in the thermal variation of polarization.

In contrast to the exhaustive measurements of various properties of ferroelectric liquid crystals at atmospheric pressure, only a few studies exist at high pressure. Here we report the isobaric thermal variation of spontaneous polarization (P(s)), coercive voltage (U(xc)), and rotational viscosity (γ(ϕ)) of a ferroelectric liquid crystal (10PPBN4) as a function of applied pressure. The material having a high value of P(s) exhibits a trend reversal: as the temperature is lowered below the transition from the smectic A to the smectic C* (ferroelectric) phase, P(s) increases to begin with but after reaching a maximum decreases with further decrease in temperature. Interestingly, the trend reversal feature becomes more dominant as the pressure is increased. Further, at a fixed reduced temperature with respect to the transition, all three parameters P(s), U(xc), and γ(ϕ) decrease with pressure. We show that the data can be well described by a model developed for materials exhibiting a sign reversal in P(s). The single characteristic parameter of the model, viz., the ratio of the inversion temperature (at which P(s) changes sign), to the transition temperature, is seen to increase with pressure, corroborating predominance of trend reversal at elevated pressures observed experimentally.

Critical behavior of three organosiloxane de Vries-type liquid crystals observed via the dielectric response.

Dielectric measurements have been made on three organosiloxane liquid crystal compounds exhibiting a smectic A (SmA) to smectic C* (SmC*) transition, the SmA phase being of the de Vries type. The electroclinic response of the molecules in the de Vries phase of these compounds exhibits a double-peak profile, and is thus different from the conventional chiral SmA phase, a feature explained on the basis of an antiferroelectric (AF) block model (Krishna Prasad et al 2009 Phys. Rev. Lett. 102 147802). The differential interactions arising from the different molecular ends of these siloxane-based compounds, which are the basis for the AF block model, can also be expected to enhance the layer translational order. We present x-ray integrated intensity data that show a high (~0.9) translational order in the SmA phase. Dielectric relaxation spectra bring out the fact that the magnitude of the soft mode relaxation parameters is dependent on the number of siloxane groups in the terminal part of the molecule. A range-shrinking analysis of the temperature-dependent dielectric relaxation strength has been carried out, using a power-law expression. The characteristic exponent shows a systematic growth with range shrinking and reaches limiting values comparable to that predicted for the 2D Ising universality class.

UV polymerization of hydrodynamically shaped fibers.

Most natural and man-made fibers have circular cross-sections; thus the properties of materials composed of non-circular fibers are largely unexplored. We demonstrate the technology for fabricating fibers with predetermined cross-sectional shape. Passive hydrodynamic focusing and UV polymerization of a shaped acrylate stream produced metre-long fibers for structural and mechanical characterization.

Critical field strength in an electroclinic liquid crystal elastomer.

We elucidate the polymer dynamics of a liquid crystal elastomer based on the time-dependent response of the pendent liquid crystal mesogens. The molecular tilt and switching time of mesogens are analyzed as a function of temperature and cross-linking density upon application of an electric field. We observe an unexpected maximum in the switching time of the liquid crystal mesogens at intermediate field strength. Analysis of the molecular tilt over multiple time regimes correlates the maximum response time with a transition to entangled polymer dynamics at a critical field strength.

Orientational order of a ferroelectric liquid crystal with small layer contraction.

We present spectroscopic and optical studies of a non-layer-shrinkage ferroelectric liquid crystal DSiKN65. The orientational order parameters S, measured with respect to the smectic layer normal using IR spectroscopy on a sample aligned homeotropically, does not exhibit any significant variation between the smectic-A∗ and smectic-C∗ phases. In contrast the birefringence of a planar homogenous sample abruptly increases at the smectic-A∗ to smectic-C∗ transition. This suggests a general increase in the orientational order, which can be described by the orientational order parameters S' defined with respect to the director. Simultaneous increase of S' and the director tilt Θ may explain the low shrinkage of smectic layers, which is consistent with recent theoretical models describing the smectic-A∗ to smectic-C∗ transition for such materials.