Light and pH responsive catanionic vesicles based on a chalcone/flavylium photoswitch for smart drug delivery: From molecular design to the controlled release of doxorubicin.

Spatially and temporally localized delivery is a promising strategy to circumvent adverse effects of traditional drug therapy such as drug toxicity and prolonged treatments. Stimuli-responsive colloidal nanocarriers can be crucial to attain such goals. Here, we develop a delivery system based on dual light and pH responsive vesicles having a cationic bis-quat gemini surfactant, 12-2-12, and a negatively charged amphiphilic chalcone, C4SCh. The premise is to exploit the chalcone/flavylium interconversion to elicit a morphological change of the vesicles leading to the controlled release of an encapsulated drug. First, the phase behavior of the catanionic system is studied and the desirable composition yielding stable unilamellar vesicles identified and selected for further studies. The solutions containing vesicles (Dh ≈ 200 nm, ζ-potential ≈ 80 mV) are in-depth characterized by light microscopy, cryo-transmission electron microscopy (cryo-TEM), dynamic light scattering (DLS) and surface tension measurements. Upon subjecting the vesicles to UV irradiation (λ = 365 nm) at near neutral pH (≈ 6.0), no morphological effects are observed, yet when irradiation is coupled with pH = 3.0, the majority of the vesicles are disrupted into bilayer fragments. The anticancer drug doxorubicin (DOX) is successfully entrapped in the non-irradiated vesicles, yielding an encapsulation efficiency of ≈ 25% and a loading capacity of ≈ 3%. The release profile of the drug-loaded vesicles is then studied in vitro in four conditions: i) no stimuli (pH = 6.0); ii) irradiation, pH = 6.0; iii) no irradiation and adjusted pH = 3.0; iv) irradiation and adjusted pH = 3.0 Crucially, irradiation at pH = 3.0 leads to a sustained release of DOX to ca. 80% (within 4 h), whereas cases i) and ii) lead to only ≈ 25 % release and case iii) to 50% release but precipitation of the vesicles. Thus, our initial hypothesis is confirmed: we present a proof of concept delivery system where light and pH act as inputs of an AND logic gate mechanism for the controlled release of a relevant biomedical drug (output). This may prove useful if the irradiated nanocarriers meet acidified physiological environments such as tumors sites, endosomes or lysosomes.

Disperse-and-Mix: Oil as an 'Entrance Door' of Carbon-Based Fillers to Rubber Composites.

Oil was employed as an 'entrance door' for loading rubber with carbon-based fillers of different size and dimensionalities: 1D carbon nanotubes (CNTs), 2D graphene nanoplatelets (GNPs), and 3D graphite. This approach was explored, as a proof of concept, in the preparation of tire tread, where oil is commonly used to reduce the viscosity of the composite mixture. Rubber was loaded with carbon black (CB, always used) and one or more of the above fillers to enhance the thermal and mechanical properties of the composite. The CNT-loaded system showed the best enhancement in mechanical properties, followed by the CNT-GNP one. Rubber loaded with both graphite and GNP showed the best enhancement in thermal conductivity (58%). The overall enhancements in both mechanical and thermal properties of the various systems were analyzed through an overall relative efficiency index in which the total filler concentration in the system is also included. According to this index, the CNT-loaded system is the most efficient one. The oil as an 'entrance door' is an easy and effective novel approach for loading fillers that are in the nanoscale and provide high enhancement of properties at low filler concentrations.

Vegetable-Oil-Based Intelligent Ink for Oxygen Sensing.

An oil-based composite is employed to monitor the exposure to oxygen inside food packaging, aiming at evaluating the package integrity and the freshness of food. The composite is an oxygen-sensitive printable ink consisting of electrically conductive silver microflakes, embedded in a vegetable oil matrix. The sensitivity of the oil to oxygen is driven by its high content of unsaturated fatty acids that polymerize and shrink upon exposure to atmospheric oxygen. Shrinkage increases the silver concentration and induces percolation, manifested by a steep increase in the electrical conductivity of the composite. We found that the electrical conductivity of the composite is related to its exposure time to air. Employing linseed oil as a matrix demonstrates an increase in electrical conductivity from 10-11 to 10-3 S/cm after only 6 days of exposure to air. We also show that this time span could be modified by changing the oil type to fit various expiration periods of food products.

Block Copolymers as Dispersants for Single-Walled Carbon Nanotubes: Modes of Surface Attachment and Role of Block Polydispersity.

When using amphiphilic polymers to exfoliate and disperse carbon nanotubes in water, the balance between the hydrophobic and hydrophilic moieties is critical and nontrivial. Here, we investigate the mode of surface attachment of a triblock copolymer, Pluronics F127, composed of a central hydrophobic polypropylene oxide block flanked by hydrophilic polyethylene oxide blocks, onto single-walled carbon nanotubes (SWNTs). Crucially, we analyze the composition in dispersant of both the as-obtained dispersion (the supernatant) and the precipitate-containing undispersed materials. For this, we combine the carefully obtained data from 1H NMR peak intensities and self-diffusion and thermogravimetric analysis. The molecular motions behind the observed NMR features are clarified. We find that the hydrophobic blocks attach to the dispersed SWNT surface and remain significantly immobilized leading to 1H NMR signal loss. On the other hand, the hydrophilic blocks remain highly mobile and thus readily detectable by NMR. The dispersant is shown to possess significant block polydispersity that has a large effect on dispersibility. Polymers with large hydrophobic blocks adsorb on the surface of the carbonaceous particles that precipitate, indicating that although a larger hydrophobic block is good for enhancing adsorption, it may be less effective in dispersing the tubes. A model is also proposed that consistently explains our observations in SWNT dispersions and some contradicting findings obtained previously in carbon nanohorn dispersions. Overall, our findings help elucidating the molecular picture of the dispersion process for SWNTs and are of interest when looking for more effective (i.e., well-balanced) polymeric dispersants.

Practical aspects in size and morphology characterization of drug-loaded nano-liposomes.

Size and morphology distributions are critical to the performance of nano-drug systems, as they determine drug pharmacokinetics and biodistribution. Therefore, comprehensive and reliable analyses of these properties are required by both the US Food and Drug Administration (FDA) and European Medicines Agency (EMA). In this study, we compare two most commonly used approaches for assessing the size distribution and morphology of liposomal nano-drug systems, namely, dynamic light scattering (DLS) and cryogenic-transmission electron microscopy (cryo-TEM); an automated quantitative analysis method was developed for the latter method. We demonstrate the advantages and disadvantages of each of these two approaches for a commercial formulation of the anti-cancer drug doxorubicin - Doxil®, in which the drug is encapsulated, mostly in the form of nano-rod crystals. With increasing drug concentration, these nano-rods change the shape of the liposomes from spherical, before drug loading, to prolate (oval), post drug loading. Cryo-TEM analysis provides a detailed size distribution of both the liposomes (minor and major axes) and the nano-rod drug. Both these values are relevant to the drug performance. In this study, we show that at elevated drug concentration (2.75 mg/ml) the drug grows mainly along the major axis and that this high concentration can result, in some cases, in liposome rupture. We show that the combination of cryo-TEM and DLS constitutes a reliable tool for demonstrating the stability of the formulation in human plasma at body temperature, a characteristic that is crucial for achieving therapeutic efficacy.

A minimal length rigid helical peptide motif allows rational design of modular surfactants.

Extensive work has been invested in the design of bio-inspired peptide emulsifiers. Yet, none of the formulated surfactants were based on the utilization of the robust conformation and self-assembly tendencies presented by the hydrophobins, which exhibited highest surface activity among all known proteins. Here we show that a minimalist design scheme could be employed to fabricate rigid helical peptides to mimic the rigid conformation and the helical amphipathic organization. These designer building blocks, containing natural non-coded α-aminoisobutyric acid (Aib), form superhelical assemblies as confirmed by crystallography and microscopy. The peptide sequence is amenable to structural modularity and provides the highest stable emulsions reported so far for peptide and protein emulsifiers. Moreover, we establish the ability of short peptides to perform the dual functions of emulsifiers and thickeners, a feature that typically requires synergistic effects of surfactants and polysaccharides. This work provides a different paradigm for the molecular engineering of bioemulsifiers.

Improving the gas barrier property of clay-polymer multilayer thin films using shorter deposition times.

Relatively fast exposure times (5 s) to aqueous solutions were found to improve the gas barrier of clay-polymer thin films prepared using layer-by-layer (LbL) assembly. Contrary to the common belief about deposition time (i.e., the longer the better), oxygen transmission rates (OTRs) of these nano-brick-wall assemblies are improved by reducing exposure time (from 1 min to 5 s). Regardless of composition, LbL films fabricated using shorter deposition time are always thicker in the first few layers, which correspond to greater clay spacing and lower OTR. A quadlayer (QL) assembly consisting of three repeat units of branched polyethylenimine (PEI), poly(acrylic acid) (PAA), PEI and montmorillonite (MMT) clay is only 24 nm thick when deposited with 1 min exposure to each ingredient. Reducing the exposure time of polyelectrolytes to 5 s not only increases this film thickness to 55 nm but also reduces the oxygen transmission rate (OTR) to 0.05 cm3/(m2 day atm), which is 2 orders of magnitude lower than the same film made using 1 min exposures. A conceptual model is proposed to explain the differences in growth and barrier, which are linked to polyelectrolyte relaxation, desorption, and interdiffusion. The universality of these findings is further exemplified by depositing clays with varying aspect ratios. This ability to quickly deposit high-barrier nanocomposite thin films opens up a tremendous opportunity in terms of commercial-scale processing of LbL assemblies.

Transient fibril structures facilitating nonenzymatic self-replication.

An emerging new direction of research focuses on developing "self-synthesizing materials", those supramolecular structures that can promote their own formation by accelerating the synthesis of building blocks and/or an entire assembly. It was postulated recently that practical design of such systems can benefit from the ability to control the assembly of amphiphilic molecules into nanostructures. We describe here the self-assembly pathway of short amphiphilic peptides into various forms of soluble ß-sheet structures--ß-plates, fibrils, and hollow nanotubes--and their consequent activity as autocatalysts for the synthesis of monomeric peptides from simpler building blocks. A detailed kinetic analysis of both the self-assembly and self-replication processes allows us to suggest a full model and simulate the replication process, revealing that only specific structures, primarily fibrils that are stable within the solution for a time shorter than a few hours, can be active as catalysts. Interestingly, we have found that such a process also induces fibril reproduction, in a mechanism very similar to the propagation of prion proteins by transmission of misfolded states.

Preparation and characterization of a novel pyrrole-benzophenone copolymerized silica nanocomposite as a reagent in a visual immunologic-agglutination test.

Biological sensing is explored through novel stable colloidal dispersions of pyrrole-benzophenone and pyrrole copolymerized silica (PPy-SiO(2)-PPyBPh) nanocomposites, which allow covalent linking of biological molecules through light mediation. The mechanism of nanocomposite attachment to a model protein is studied by gold labeled cholera toxin B (CTB) to enhance the contrast in electron microscopy imaging. The biological test itself is carried out without gold labeling, i.e., using CTB only. The protein is shown to be covalently bound through the benzophenone groups. When the reactive PPy-SiO(2)-PPyBPh-CTB nanocomposite is exposed to specific recognition anti-CTB immunoglobulins, a qualitative visual agglutination assay occurs spontaneously, producing as a positive test, PPy-SiO(2)-PPyBPh-CTB-anti-CTB, in less than 1 h, while the control solution of the PPy-SiO(2)-PPyBPh-CTB alone remained well-dispersed during the same period. These dispersions were characterized by cryogenic transmission microscopy (cryo-TEM), scanning electron microscopy (SEM), FTIR and X-ray photoelectron spectroscopy (XPS).