Ferulic acid-loaded polymeric nanoparticles prepared from nano-emulsion templates facilitate internalisation across the blood-brain barrier in model membranes.

A hydroxycinnamic acid derivative, namely ferulic acid (FA) has been successfully encapsulated in polymeric nanoparticles (NPs) based on poly(lactic-co-glycolic acid) (PLGA). FA-loaded polymeric NPs were prepared from O/W nano-emulsion templates using the phase inversion composition (PIC) low-energy emulsification method. The obtained PLGA NPs exhibited high colloidal stability, good drug-loading capacity, and particle hydrodynamic diameters in the range of 74 to 117 nm, depending on the FA concentration used. In vitro drug release studies confirmed a diffusion-controlled mechanism through which the amount of released FA reached a plateau at 60% after 6 hours-incubation. Five kinetic models were used to fit the FA release data as a function of time. The Weibull distribution and Korsmeyer-Peppas equation models provided the best fit to our experimental data and suggested quasi-Fickian diffusion behaviour. Moderate dose-response antioxidant and radical scavenging activities of FA-loaded PLGA NPs were demonstrated using the DPPH˙ assay achieving inhibition activities close to 60 and 40%, respectively. Cell culture studies confirmed that FA-loaded NPs were not toxic according to the MTT colorimetric assay, were able to internalise efficiently SH-SY5Y neuronal cells and supressed the intracellular ROS-level induced by H2O2 leading to 52% and 24.7% of cellular viability at 0.082 and 0.041 mg mL-1, respectively. The permeability of the NPs through the blood brain barrier was tested with an in vitro organ-on-a-chip model to evaluate the ability of the FA-loaded PLGA and non-loaded PLGA NPs to penetrate to the brain. NPs were able to penetrate the barrier, but permeability decreased when FA was loaded. These results are promising for the use of loaded PLGA NPs for the management of neurological diseases.

Polymer nanoparticles from low-energy nanoemulsions for biomedical applications.

The formulation of nanoemulsions by low-energy strategies, particularly by the phase inversion composition method, and the use of these nanoemulsions as templates for the preparation of polymer nanoparticles for biomedical applications are reviewed. The methods of preparation, nature of the components in the formulation, and their impact on the physicochemical properties, drug loading, and drug release are discussed. We highlight the utilization of ethyl cellulose, poly(lactic-co-glycolic acid), and polyurethane/polyurea in the field of nanomedicine as potential drug delivery systems. Advances are still needed to achieve better control over size distribution, nanoparticle concentration, surface functionalization, and the type of polymers that can be processed.

Rosmarinic Acid-Loaded Polymeric Nanoparticles Prepared by Low-Energy Nano-Emulsion Templating: Formulation, Biophysical Characterization, and In Vitro Studies.

Rosmarinic acid (RA), a caffeic acid derivative, has been loaded in polymeric nanoparticles made up of poly(lactic-co-glycolic acid) (PLGA) through a nano-emulsion templating process using the phase-inversion composition (PIC) method at room temperature. The obtained RA-loaded nanoparticles (NPs) were colloidally stable exhibiting average diameters in the range of 70-100 nm. RA was entrapped within the PLGA polymeric network with high encapsulation efficiencies and nanoparticles were able to release RA in a rate-controlled manner. A first-order equation model fitted our experimental data and confirmed the prevalence of diffusion mechanisms. Protein corona formation on the surface of NPs was assessed upon incubation with serum proteins. Protein adsorption induced an increase in the hydrodynamic diameter and a slight shift towards more negative surface charges of the NPs. The radical scavenging activity of RA-loaded NPs was also studied using the DPPH·assay and showed a dose-response relationship between the NPs concentration and DPPH inhibition. Finally, RA-loaded NPs did not affect the cellular proliferation of the human neuroblastoma SH-SY5Y cell line and promoted efficient cellular uptake. These results are promising for expanding the use of O/W nano-emulsions in biomedical applications.

Chromonic nematic liquid crystals in a room-temperature ionic liquid.

Planar multiaromatic molecules hierarchically and selectively arrange into nematic chromonic liquid crystals in the room temperature ionic liquid 2-hydroxyethylammonium formate. In a proof of concept, these liquid crystals were used as reaction media to produce mesostructured silica materials under mild biomimetic conditions. Several other applications are envisaged.

A nanocellulose-based platform towards targeted chemo-photodynamic/photothermal cancer therapy.

Cellulose nanocrystals (CNCs) have advantages as drug delivery carriers because of their biocompatibility and the presence of hydroxyl groups which favor chemical modification and drug binding. The present study describes the development of novel multifunctional rod-like CNCs-based carriers as therapeutic platforms: CNCs were hybridized with folic acid for actively targeting tumor cells, carbon dots (Cdots) for both imaging and photodynamic/photothermal treatments and doxorubicin (DOX) as an anticancer drug. Hybridized carriers displayed excellent drug-loading capacity. Moreover, Cdots-containing hybrids showed fluorescence and photosensitized singlet oxygen generation and photothermal behavior. Carriers exhibited pH-sensitive drug release because of changing interactions with DOX, and this release proved to be effective against in vitro cervical cancer cells, as evidenced by dose-dependent reduced cellular viabilities. Additionally, DOX release was promoted by light irradiation and the photodynamic behavior by reactive oxygen species was confirmed. These results demonstrate the potential of multifunctional CNCs-based carriers as platforms for multimodal photodynamic/photothermal-chemotherapy.

1D materials from ionic self-assembly in mixtures containing chromonic liquid crystal mesogens.

Ionic self-assembly is a simple yet powerful method to obtain robust nanostructures. Herewith, we use mixtures of oppositely-charged porphyrins that can act as mesogens to form chromonic liquid crystals in water, i.e., molecular stacks with orientational (nematic) or positional (hexagonal) order. Electrostatic locking coupled with π-π interactions between aromatic groups within the stacks, together with inter-stack hydrogen bonding induce formation of all-organic crystalline nanofibers with high aspect ratio (a few tenths of nanometers in width but several tenths of micrometers in length) and that display branching. The nanofibers prepared from metal-free porphyrin units feature interesting optical properties, including an absorption spectrum that is different from the simple sum of the individual spectra of the components, which is attributed to a striking aggregation-induced chromism. When in contact with some polar organic solvents the materials become fluorescent, as a result of disaggregation. In a proof-of-concept, the obtained self-assembled one-dimensional (1D) materials were carbonized (yield ca. 60%) to produce nitrogen-doped carbon nanofibers that can be used as active electrode materials for energy storage applications.

Functionalized PLGA nanoparticles prepared by nano-emulsion templating interact selectively with proteins involved in the transport through the blood-brain barrier.

During the last few decades, extensive efforts has been made to design nanocarriers to transport drugs into the central nervous system (CNS). However, its efficacy is limited due to the presence of the Blood-Brain Barrier (BBB) which greatly reduces drug penetration making Drug Delivery Systems (DDS) necessary. Polymeric nanoparticles (NPs) have been reported to be appropriate for this purpose and in particular, poly(lactic-co-glycolic acid) (PLGA) has been used for its ability to entrap small molecule drugs with great efficiency and the ease with which it functionalizes NPs. Despite the fact that their synthetic identity has been studied in depth, the biological identity of such manufactured polymers still remains unknown as does their biodistribution and in vivo fate. This biological identity is a result of their interaction with blood proteins, the so-called "protein corona" which tends to alter the behavior of polymeric nanoparticles in the body. The aim of the present research is to identify the proteins bounded to polymeric nanoparticles designed to selectively interact with the BBB. For this purpose, four different PLGA NPs were prepared and analyzed: (i) "PLGA@Drug," in which a model drug was encapsulated in its core; (ii) "8D3-PLGA" NPs where the PLGA surface was functionalized with a monoclonal anti-transferrin receptor antibody (8D3 mAb) in order to specifically target the BBB; (iii) "8D3-PLGA@Drug" in which the PLGA@Drug surface was functionalized using the same antibody described above and (iv) bare PLGA NPs which were used as a control. Once the anticipated protein corona NPs were obtained, proteins decorating both bare and functionalized PLGA NPs were isolated and analyzed. Apart from the indistinct interaction with PLGA NPs with the most abundant serum proteins, specific proteins could also be identified in the case of functionalized PLGA NPs. These findings may provide valuable insight into designing novel vehicles based on PLGA NPs for crossing the BBB.

Short and ultrashort antimicrobial peptides anchored onto soft commercial contact lenses inhibit bacterial adhesion.

Commercial soft contact lenses were chemically modified to incorporate antibacterial properties. Contact lenses and especially soft contact lenses present a risk of eye microbial infection that eventually may lead to vision loss. This is a significant health issue given the large population of contact lenses wearers worldwide. In order to introduce bactericidal activity in hydrogel contact lenses, one short and one ultrashort antimicrobial peptides, LKKLLKLLKKLLKL (LK) and IRIRIRIR (IR), were selected. These peptides were anchored on the surface of contact lenses using a linker (1,4-butanediol diglycidyl ether) under mild conditions (room temperature, pH = 7.4). Physical and chemical properties of peptide-functionalized contact lenses were investigated through several analytical techniques including wettability, Raman confocal microscopy, fluorescence studies, refractometry and spectrophotometry. These studies demonstrated that contact lens modification occurred at the nanolevel (ng/lens). Bacterial cultures showed that peptide-functionalized contact lenses can drastically reduce bacterial adhesion and viability when exposed to Pseudomonas aeruginosa and Staphylococcus aureus. These systems offer the potential to minimise corneal bacterial infection and represent a suitable platform for future ophthalmic devices.

Biomedical perfluorohexane-loaded nanocapsules prepared by low-energy emulsification and selective solvent diffusion.

Perfluorohexane-loaded nanocapsules are interesting materials for many biomedical applications such as oxygen delivery systems or contrast agents. However, their formulation into stable colloidal systems is challenging because of their hydro- and lipophobicity, high density and high vapour pressure. In this study, perfluorohexane-loaded polymeric nanocapsules are prepared for the first time by low-energy emulsification and selective solvent diffusion. The colloidal stability of the perfluorohexane nano-emulsion templates has been improved by the incorporation of an apolar low-density oil (isopropyl myristate) in the dispersed phase, thus addressing droplet coarsening and migration phenomena. The perfluorohexane-loaded nanocapsules prepared from the nano-emulsions show sizes smaller than the corresponding emulsion templates (below 150 nm by dynamic light scattering) and exhibit good stability under storage conditions. Hyperspectral enhanced dark field microscopy revealed a layered core/shell structure and allowed also to confirm the encapsulation of perfluorohexane which was quantified by elemental microanalysis. Although isopropyl myristate has an unfavourable biocompatibility profile, cell viability is enhanced when perfluorohexane is present in the nanocapsules, which is attributed to its high oxygen transport capacity.

Cubic liquid crystalline structures in diluted, concentrated and highly concentrated emulsions for topical application: Influence on drug release and human skin permeation.

Novel emulsions with a nanostructured continuous phase have been proposed as controlled drug delivery systems to enhance topical delivery of active ingredients avoiding systemic effects. In this study, oil-in-water (O/W) emulsions with two surfactant/water (S/W) weight ratios of 40:60 and 35:65, and oil concentrations of 10 wt% (diluted emulsion), 40 wt% (concentrated emulsion) and 85 wt% (highly concentrated emulsion) have been investigated to identify the presence of liquid crystalline structures and their influence on drug release and skin permeation. The emulsions have been characterized in terms of visual appearance, rheology and drug release. The presence of cubic liquid crystalline structures in emulsions with S/W 40:60 was confirmed by small angle X-ray scattering (SAXS). Rheology results showed a markedly different behaviour in emulsions with S/W 40:60 compared with nonstructured emulsions. A model drug, diclofenac sodium (DS) was successfully incorporated in the emulsions. DS release was studied with hydrophilic and lipophilic membranes, and the amount of DS in the receptor solution was significantly lower in the formulations containing cubic liquid structures. An in vitro skin permeation study with dermatomed human skin showed that emulsions with a nanostructured continuous phase are suitable formulations for topical delivery with DS retention in skin layers. The results indicate that the amount of drug retained in skin structures may be tuned by modification of liquid crystal concentration and emulsion structure.

Formulating stable hexosome dispersions with a technical grade diglycerol-based surfactant.

We report on the phase behavior of a technical grade and commercially available diglycerol monoisostearate, C41V, and its use for the preparation of nanostructured liquid crystal dispersions (hexosomes). C41V in water forms a reverse hexagonal liquid crystal at room temperature and in a wide range of concentrations (0.5-95 wt%); this hexagonal liquid crystal is stable up to 70 °C. A simple and effective method has been developed to disperse hexosomes with an encapsulated active molecule (Ketoprofen) that consists of (1) producing a nano-emulsion stabilized by an amphiphilic block copolymer (Pluronic F127) and containing ethyl acetate and C41V by using ultrasounds and (2) evaporating the solvent to produce hexosomes. The size of the hexosomes and ultrasound dispersion time is markedly reduced by using ethyl acetate as an auxiliary solvent with an optimal initial ratio of C41V:ethyl acetate of 50:50. Dynamic light scattering shows that the size of the hexosomes decreases as the concentration of stabilizer F127 or encapsulated Ketoprofen is increased. The lattice parameter in the hexagonal structure is calculated from small angle scattering data to be ca. 5.3  nm and is only slightly dependent on the amount of F127 and/or encapsulated Ketoprofen. Cryo electron microscopy reveals that the samples contain hexosomes and these coexist with spherical, likely F127 micelles. Lastly, hexosomes show a pH responsive release of Ketoprofen which could be useful for target delivery in the gastrointestinal tract.

Template-directed self-organization of colloidal PbTe nanocrystals into pillars, conformal coatings, and self-supported membranes.

We demonstrate the formation of three morphologies relevant for integration with miniaturized devices-microscale pillars, conformal coatings, and self-supported membranes-via template-directed self-organization of lead telluride (PbTe) colloidal nanocrystals (NCs). Optimizing the self-organization process towards producing one of these morphologies typically involves adjusting the surface chemistry of the particles, as a means of controlling the particle-particle and particle-template interactions. In contrast, we have produced each of the three morphologies of close-packed NCs by adjusting only the solvent and concentration of NCs, to ensure that the high quality of the ca. 10 nm PbTe NCs produced by hot-injection colloidal synthesis, which we used as model "building blocks," remains consistent across all three configurations. For the first two morphologies, the NCs were deposited as colloidal suspensions onto micropatterned silicon substrates. The microscale cuboid pillars (1 µm × 1 µm × 0.6 µm) were formed by depositing NC dispersions in toluene onto templates patterned with resist grid motifs, followed by the resist removal after the slow evaporation of toluene and formation of the micropillars. Conformal coatings were produced by switching the solvent from toluene to a faster drying hexane and pouring NC dispersions onto silicon templates with topographically patterned microstructures. In a similar process, self-supported NC membranes were formed from NC dispersions in hexane on the surface of diethylene glycol and transferred onto the micropatterned templates. The demonstrated combination of bottom-up self-organization with top-down micropatterned templates provides a scalable route for design and fabrication of NC ensembles in morphologies and form-factors that are compatible with their integration into miniaturized devices.

Development of Inhalable Superparamagnetic Iron Oxide Nanoparticles (SPIONs) in Microparticulate System for Antituberculosis Drug Delivery.

Tuberculosis (TB) is an infectious disease which affects millions of people worldwide. Inhalable polymeric dry powders are promising alternatives as anti-TB drug carriers to the alveoli milieu and infected macrophages, with potential to significantly improve the therapeutics efficiency. Here, the development of a magnetically responsive microparticulate system for pulmonary delivery of an anti-TB drug candidate (P3) is reported. Microparticles (MPs) are developed based on a cast method using calcium carbonate sacrificial templates and incorporate superparamagnetic iron oxide nanoparticles to concentrate MPs in alveoli and enable drug on demand release upon actuation of an external alternate magnetic field (AMF). The MPs are shown to be suitable for P3 delivery to the lower airways and for alveolar macrophage phagocytosis. The developed MPs reveal unique and promising features to be used as an inhalable dry powder allowing the AMF control over dosage and frequency of drug delivery anticipating improved TB treatments.

Adsorption of marine phycotoxin okadaic acid on a covalent organic framework.

Phycotoxins, compounds produced by some marine microalgal species, can reach high concentrations in the sea when a massive proliferation occurs, the so-called harmful algal bloom. These compounds are especially dangerous to human health when concentrated in the digestive glands of seafood. In order to generate an early warning system to alert for approaching toxic outbreaks, it is very important to improve monitoring methods of phycotoxins in aquatic ecosystems. Solid-phase adsorption toxin tracking devices reported thus far based on polymeric resins have not been able to provide an efficient harmful algal bloom prediction system due to their low adsorption capabilities. In this work, a water-stable covalent organic framework (COF) was evaluated as adsorbent for the hydrophobic toxin okadaic acid, one of the most relevant marine toxins and the parental compound of the most common group of toxins responsible for the diarrhetic shellfish poisoning. Adsorption kinetics of okadaic acid onto the COF in seawater showed that equilibrium concentration was reached in only 60min, with a maximum experimental adsorption of 61mgg-1. Desorption of okadaic acid from the COF was successful with both 70% ethanol and acetonitrile as solvent, and the COF material could be reused with minor losses in adsorption capacity for three cycles. The results demonstrate that COF materials are promising candidates for solid-phase adsorption in water monitoring devices.

A colloidally stable water dispersion of Ni nanowires as an efficient T2-MRI contrast agent.

A colloidally stable dispersion of anisotropic Ni nanowires in water has been achieved showing good performance as a T2-contrast agent in magnetic resonance imaging (MRI). A wet chemistry approach has been developed that renders water dispersible Ni nanowires of controlled size. Firstly, an array of Ni/Au multilayer nanowires of tailored dimensions was synthesized by pulsed electrodeposition within a porous alumina template in a three-electrode cell at room-temperature. Then, Ni/Au multilayer nanowires were released from the template followed by the isolation of the Ni segments through a two-step acidic etching, ending up with a colloidally stable dispersion of poly-acrylic acid (PAA)-coated Ni nanowires in water solution. The PAA-coated Ni nanowires have been morphologically and structurally characterized by scanning and transmission electron microscopy (SEM, TEM, respectively), whereas their ferromagnetic properties were studied using a SQUID-magnetometer. Importantly, relaxivity properties (transversal relaxivity, r2) indicated good performance of PAA-coated Ni nanowires as a T2-contrast agent at 1.41 T and body temperature, with values as large as those of commercial contrast agents based on iron oxide nanoparticles. MRI phantom imaging at clinical fields of 3 T confirmed the dark contrast effect of PAA-coated Ni nanowires, as compared to a water control, which opens a new window of opportunities for anisotropic metal nanostructures as contrast agents in MRI applications.

From Chromonic Self-Assembly to Hollow Carbon Nanofibers: Efficient Materials in Supercapacitor and Vapor-Sensing Applications.

Carbon nanofibers (CNFs) with high surface area (820 m2/g) have been successfully prepared by a nanocasting approach using silica nanofibers obtained from chromonic liquid crystals as a template. CNFs with randomly oriented graphitic layers show outstanding electrochemical supercapacitance performance, exhibiting a specific capacitance of 327 F/g at a scan rate of 5 mV/s with a long life-cycling capability. Approximately 95% capacitance retention is observed after 1000 charge-discharge cycles. Furthermore, about 80% of capacitance is retained at higher scan rates (up to 500 mV/s) and current densities (from 1 to 10 A/g). The high capacitance of CNFs comes from their porous structure, high pore volume, and electrolyte-accessible high surface area. CNFs with ordered graphitic layers were also obtained upon heat treatment at high temperatures (>1500 °C). Although it is expected that these graphitic CNFs have increased electrical conductivity, in the present case, they exhibited lower capacitance values due to a loss in surface area during thermal treatment. High-surface-area CNFs can be used in sensing applications; in particular, they showed selective differential adsorption of volatile organic compounds such as pyridine and toluene. This behavior is attributed to the free diffusion of these volatile aromatic molecules into the pores of CNFs accompanied by interactions with sp2 carbon structures and other chemical groups on the surface of the fibers.

EGFR-Based Immunoisolation as a Recovery Target for Low-EpCAM CTC Subpopulation.

Circulating tumour cells (CTCs) play a key role in the metastasis process, as they are responsible for micrometastasis and are a valuable tool for monitoring patients in real-time. Moreover, efforts to develop new strategies for CTCs isolation and characterisation, and the translation of CTCs into clinical practice needs to overcome the limitation associated with the sole use of Epithelial Cell Adhesion Molecule (EpCAM) expression to purify this tumour cell subpopulation. CTCs are rare events in the blood of patients and are believed to represent the epithelial population from a primary tumour of epithelial origin, thus EpCAM immunoisolation is considered an appropriate strategy. The controversy stems from the impact that the more aggressive mesenchymal tumour phenotypes might have on the whole CTC population. In this work, we first characterised a panel of cell lines representative of tumour heterogeneity, confirming the existence of tumour cell subpopulations with restricted epithelial features and supporting the limitations of EpCAM-based technologies. We next developed customised polystyrene magnetic beads coated with antibodies to efficiently isolate the phenotypically different subpopulations of CTCs from the peripheral blood mononuclear cells (PBMCs) of patients with metastatic cancer. Besides EpCAM, we propose Epidermal Growth Factor Receptor (EGFR) as an additional isolation marker for efficient CTCs detection.

A supramolecular strategy based on molecular dipole moments for high-quality covalent organic frameworks.

A supramolecular strategy based on strong molecular dipole moments is presented to gain access to covalent organic framework structures with high crystallinity and porosity. Antiparallel alignment of the molecules within the pore walls is proposed to lead to reinforced columnar stacking, thus affording a high-quality material. As a proof of principle, a novel pyrene dione building block was prepared and reacted with hexahydroxytriphenylene to form a boronic ester-linked covalent organic framework. We anticipate the strategy presented herein to be valuable for producing highly defined COF structures.

Up-scaling the synthesis of Cu2O submicron particles with controlled morphologies for solar H2 evolution from water.

The synthesis of Cu2O was studied to examine the effects of up-scaling on the size and morphology of the resultant particles. As a result, a successful protocol employing an automated laboratory reactor was developed for large-scale synthesis of phase-pure Cu2O colloids with specific sizes in the submicron to micrometer range (0.2-2.6 µm). The as-synthesized products have been studied by means of powder X-ray diffraction, Raman spectroscopy, X-ray photoelectron spectroscopy, UV-Vis-NIR spectroscopy, scanning electron microscopy, and photoelectrochemical measurements. A broad range of morphologies, both equilibrium (stellated octahedrons, cubes, cuboctahedrons, truncated octahedrons, truncated cuboctahedrons) and metastable (cage-like hierarchical structures, microspheres with flower-like texture), with uniform sizes have been selectively prepared either by careful tuning of synthesis conditions. Recrystallization of primary aggregates through Ostwald ripening is proposed as the formation mechanism for these Cu2O structures. As a photocathode for photoelectrochemical H2 evolution, Cu2O submicron cubes with exposed {001} facets exhibit a high open-circuit potential of ca. 0.9 V vs. the RHE at pH 1.

Multiple emulsions as soft templates for the synthesis of multifunctional silicone porous particles.

Multiple emulsion templating is a versatile strategy for the synthesis of porous particles. The present work addresses the synthesis of multifunctional poly(dimethylsiloxane) porous particles using multiple water-in-oil-in-water emulsions as soft templates with an oil phase constituted by a crosslinkable poly(dimethylsiloxane) (PDMS) oil. Herewith, the impact of the viscosity of PDMS oil (i.e., molecular weight) on the properties of both the emulsion templates and the resulting particles was evaluated. The viscosity of PDMS oil has a strong effect on the size and polydispersity of the emulsion templates as well as on the mechanical properties of the derived particles. The elastic modulus can be tuned by mixing PDMS oils of different viscosities to form bimodal crosslinked networks. Iron oxide nanoparticles can be readily incorporated into the emulsion templates to provide additional functionalities to the silicone particles, such as magnetic separation or magnetic hyperthermia. The synthesized composite magnetic particles were found to be useful as recoverable absorbent materials (e.g., for oil spills) by taking advantage of their high buoyancy and high hydrophobicity.