Polymeric Nanocontainers Research Articles

Thermoresponsive supramolecular nanocontainers from ionic complexes of amphiphilic wedge-shaped mesogens and polybases

Multi-responsive polymeric nanocontainers attract significant attention for their potential applications in biotechnology, drug delivery, catalysis, and other fields. By incorporating a liquid-crystalline (LC) mesogenic ligand with an alkyl tail length ranging from 8–12 carbons, ionically linked to the polymer backbone, we generate vesicles with walls significantly thinner than those of conventional polymersomes, approaching the thickness of a lipid bilayer. These LC vesicles, ranging in size from 50–120 nm, are designed to be mechanically robust due to the alignment of the hydrophilic polymer backbone within the plane of the vesicle wall. Additionally, incorporating a temperature-sensitive block into the polymer structure imparts thermoresponsiveness to the nanocontainers, enhancing their functionality and adaptability for various applications. Ionic complexes of hydrophilic polybases, specifically poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) and PDMAEMA-b-PNIPAM (poly(N-isopropylacrylamide)) block copolymers, with amphiphilic wedge-shaped mesogens bearing a sulfonic acid group at the focal point were synthesized. The designed nanocontainers, in the form of either vesicles or nanotubes, exhibit a well-defined wall thickness of 5 nm, dictated by the organization of a smectic LC phase. The constructed coarse-grained models elucidate the mechanism of self-assembly, demonstrating that the balance between the hydrophilicity of the main polymer chain and the hydrophobicity of the wedge-shaped pendant groups determines both the internal and external structure of the vesicles.

Hybrid Zinc Coating with CuO Nanocontainers Containing Corrosion Inhibitor for Combined Protection of Mild Steel from Corrosion and Biofouling

In this study, a multifunctional hybrid coating is designed for the combined protection of mild steel from corrosion and biofouling in aggressive salt water. This involves preparation of a pH-responsive-release system based on copper oxide (CuO) as a biocide, and the corrosion inhibitor Safranin loaded in polymeric nanocontainers by alternate adsorption of poly(acrylic acid) and poly(ethylenimine) on CuO nanoparticles in water solutions. By optimizing the conditions, i.e., pH and concentration, good stability of the suspensions and the loading amount of Safranin is achieved. The nanocontainers are electrodeposited as an intermediate layer in an ordinary zinc coating on steel (“sandwich-like” structure) from the water solution in order to minimize the effect of CuO dissolution. To highlight the role of Safranin in reducing steel corrosion, a second zinc coating containing CuO nanoparticles without a corrosion inhibitor is also examined. The surface morphology and corrosion behavior of the hybrid coatings are evaluated in a model corrosion medium (5% NaCl solution). Both coatings are found to improve the anticorrosion behavior of steel for a time interval of 55 days and at conditions of external polarization. It can be expected that the newly developed hybrid coatings would also demonstrate potential for marine applications due to the main characteristics of their components.

Mathematical modelling of drug delivery from pH-responsive nanocontainers

Targeted drug delivery systems represent a promising strategy to treat localised disease with minimum impact on the surrounding tissue. In particular, polymeric nanocontainers have attracted major interest because of their structural and morphological advantages and the variety of polymers that can be used, allowing the synthesis of materials capable of responding to the biochemical alterations of the environment. While experimental methodologies can provide much insight, the generation of experimental data across a wide parameter space is usually prohibitively time consuming and/or expensive. To better understand the influence of varying design parameters on the release profile and drug kinetics involved, appropriately-designed mathematical models are of great benefit. Here, we developed a continuum-scale mathematical model to describe drug transport within, and release from, a hollow nanocontainer consisting of a core and a pH-responsive polymeric shell. Our two-layer mathematical model accounts for drug dissolution and diffusion and includes a mechanism to account for trapping of drug molecules within the shell. We conduct a sensitivity analysis to assess the effect of varying the model parameters on the overall behaviour of the system. To demonstrate the usefulness of our model, we focus on the particular case of cancer treatment and calibrate the model against release profile data for two anti-cancer therapeutical agents. We show that the model is capable of capturing the experimentally observed pH-dependent release.

Multisensitive Polymeric Nanocontainers as Drug Delivery Systems: Biological Evaluation.

This chapter focuses on the in vitro biological evaluation of multisensitive nanocontainers as drug delivery systems for cancer treatment. Cancer tissues possess some unique characteristics such as increased temperature due to inflammation, thermal vulnerability (40-45°C), low cellular pH, and redox instabilities. The employment of polymers bearing pH, thermo, and/or redox sensitivities in the synthesis of hollow polymeric nanostructures has led to the formulation of a variety of drug delivery vehicles that are capable of targeted delivery and trigger specific drug release. The cavity in the structure allows for the encapsulation of anticancer drugs as well as other moieties with anticancer activity, like iron oxide magnetic nanoparticles. The drug loading and release capability of the nanocontainers is evaluated prior to biological studies in order to determine the concentration of the drug in the structure. The in vitro assessment includes cytotoxicity studies, quantitatively through the colorimetric MTT assay as well as qualitatively via the scratch-wound healing assay, on both cancer and healthy cell lines. The cellular localization of the studied drug-loaded and unloaded nanocontainers is determined through confocal fluorescence microscopy.

P.412 Gallic acid, chlorogenic acid, ferulic acid and quercetin improve learning and memory processes in olfactory bulbectomized rats

Polymeric nanocontainers (NCs) impregnated with corrosion inhibitor benzotriazole are formed using layer-by-layer deposition of poly(acrylic acid) and poly(diallyldimethylammonium chloride) onto hematite particles in the presence of 0.1 M NaCl. Electric light scattering and electrophoresis are employed to control the formation and the size of NCs, as well as the stability of their suspensions. The inhibitor loaded NCs are incorporated into the volume of the galvanized coating during the electrodeposition of zinc on the steel substrate to ensure additional self-healing effect in the case of corrosion attack. The surface morphology and the uniformity of this composite coating as well as the lack of aggregation of the nanocontainers is demonstrated by scanning electron microscopy. The influence of NCs present in the zinc electrolyte on the cathodic and anodic processes is investigated by cyclic voltammetry. The corrosion behavior of the composite coatings at conditions of external cathodic and anodic polarization is tested with potentiodynamic measurements and the results are compared to pure zinc coatings. The composite coatings with embedded NCs revealed enhanced corrosion protection of low carbon steel in comparison with the pure zinc coatings in neutral corrosion medium.

A facile design of smart silica nanocarriers via surface-initiated RAFT polymerization as a dual-stimuli drug release platform

The rational design of hybrid nanostructures with an active core and a modified shell can promote several biological applications. Herein, a coating platform has been developed to obtain monodispersed core-shell magnetic mesoporous silica nanocarriers. The surface of silica shells has been successfully modified by disulfide bonds and the photoresponsive polymer, poly(2-nitrobenzyl acrylate) (PNBA), via RAFT polymerization using azobisisobutyronitrile (AIBN) as a thermal initiator. The results showed that these nanocarriers have nanospherical morphologies with high porosity, large surface areas, and suitable pore volumes. Furthermore, ibuprofen (IBU) as a model drug has been loaded easily into the polymeric nanocontainers with a high loading content, followed by coating with β-Cyclodextrin (β-CD) as effective gatekeeper molecules. Treating these nanocarriers with ultraviolet light causes the photocleavage of o-nitrobenzyl ester moieties contained in the growing polymer chains. The controlled release of the loaded IBU cargo can be achieved by ultraviolet light, or reduction cleavable of the disulfide bond, or both together. Interestingly, the magnetic core can enhance the release process through magnetic guiding and pushing the nanocarriers to the targeted sites by using an external magnet or a directed magnetic field. The prepared photodegradable polymer nanocarriers can be widely applied in the fields of controlled drug delivery systems (DDS) and pharmacologically active polymers.

Gold nanoparticle decorated pH-sensitive polymeric nanocontainers as a potential theranostic agent

A pH-sensitive system of hollow P(MAA-co-MBA-co-AA) nanocontainers (NCs) modified with gold nanoparticles (GNCs) has been developed for theranostic applications, drug delivery and real time monitoring through imaging. The GNCs were synthesised by the distillation precipitation copolymerization procedure followed by in situ synthesis and embodiment of gold nanoparticles on the polymeric matrix (CSNs). Separately, citrate capped gold nanoparticles (GNPs) were also synthesized and compared with the GNCs for their fluorescence and cellular localization ability. The GNCs were tested for their drug loading and release behavior in response to the anticancer drug doxorubicin (DOX) at different pH values. Sustained drug release was observed at acidic pH. The viability of MCF-7 breast cancer cells and HEK-293 human embryonic kidney cells in relation to the GNCs, GNPs, GNC@DOX and DOX was also evaluated. GNCs and GNPs at the tested concentrations did not inhibit proliferation at either cell lines, whereas the GNCs@DOX presented comparable results. Cellular migration of MCF-7 cells treated with GNCs or GNPs was evaluated through the scratch-wound healing assay but no significant inhibition was detected. GNCs’ and GNPs’ fluorescence ability was exploited for assessing cellular localization through confocal laser scanning microscopy. GNCs after only 1 h of treatment were found in the cytoplasm of MCF7 cells, whereas GNCs@DOX were localized in the nucleus; the desirable site of action of DOX.

Serum amyloid A protein as a diagnostic marker for viral infections

Polymeric nanocontainers (NCs) impregnated with corrosion inhibitor benzotriazole are formed using layer-by-layer deposition of poly(acrylic acid) and poly(diallyldimethylammonium chloride) onto hematite particles in the presence of 0.1 M NaCl. Electric light scattering and electrophoresis are employed to control the formation and the size of NCs, as well as the stability of their suspensions. The inhibitor loaded NCs are incorporated into the volume of the galvanized coating during the electrodeposition of zinc on the steel substrate to ensure additional self-healing effect in the case of corrosion attack. The surface morphology and the uniformity of this composite coating as well as the lack of aggregation of the nanocontainers is demonstrated by scanning electron microscopy. The influence of NCs present in the zinc electrolyte on the cathodic and anodic processes is investigated by cyclic voltammetry. The corrosion behavior of the composite coatings at conditions of external cathodic and anodic polarization is tested with potentiodynamic measurements and the results are compared to pure zinc coatings. The composite coatings with embedded NCs revealed enhanced corrosion protection of low carbon steel in comparison with the pure zinc coatings in neutral corrosion medium.

Development of multi-layered and multi-sensitive polymeric nanocontainers for cancer therapy: in vitro evaluation

Nanoscale drug delivery systems represent a promising strategy to treat cancer and to overcome the side effects of chemotherapy. In particular, hollow polymeric nanocontainers have attracted great interest because of their structural and morphological advantages and the variety of polymers that can be used, allowing the synthesis of stimuli-responsive materials capable of responding to the biochemical alterations of the tumour microenvironment. Here are reported the synthesis, characterization and in vitro evaluation of a three-stimuli-sensitive hollow nanocontainer consisting of three different shells, each one sensitive to a specific tumoral stimulus: in order pH, temperature and reducing environment. To test its properties, daunorubicin was used as a model drug, for which the nanocontainers exhibited excellent encapsulation ability. The in vitro drug release behaviour was studied under different conditions, where the system proved capable of responding to the selected tumoral stimuli by releasing a larger amount of drug than in physiological environment. The hollow system itself showed negligible cytotoxicity but the loaded nanocontainers and free drug showed identical cytotoxicity and intracellular localization. Therefore, this formulation can be considered as a promising platform to develop an injectable delivery system capable of improving systematic toxicity without affecting or reducing the activity of the encapsulated drug.

Immobilization of Benzocaine in Polymeric Nanocontainers. Pharmacokinetic Modeling

The possibility of immobilizing benzocaine in polymeric nanocontainers made of sulfonated cation exchangers, i.e., sulfonated polycalixarene and KU-23 30/100, was studied. The desorption kinetics of neutral and cationic benzocaine from the polymeric nanocontainers provided the desired drug-release pharmacokinetics upon peroral administration.

Design of polymeric core-shell nanocontainers impregnated with benzotriazole for active corrosion protection of galvanized steel

Abstract Polymeric nanocontainers (NCs) impregnated with corrosion inhibitor benzotriazole are formed using layer-by-layer deposition of poly(acrylic acid) and poly(diallyldimethylammonium chloride) onto hematite particles in the presence of 0.1 M NaCl. Electric light scattering and electrophoresis are employed to control the formation and the size of NCs, as well as the stability of their suspensions. The inhibitor loaded NCs are incorporated into the volume of the galvanized coating during the electrodeposition of zinc on the steel substrate to ensure additional self-healing effect in the case of corrosion attack. The surface morphology and the uniformity of this composite coating as well as the lack of aggregation of the nanocontainers is demonstrated by scanning electron microscopy. The influence of NCs present in the zinc electrolyte on the cathodic and anodic processes is investigated by cyclic voltammetry. The corrosion behavior of the composite coatings at conditions of external cathodic and anodic polarization is tested with potentiodynamic measurements and the results are compared to pure zinc coatings. The composite coatings with embedded NCs revealed enhanced corrosion protection of low carbon steel in comparison with the pure zinc coatings in neutral corrosion medium.

Tailor-made nanocontainers for combined magnetic-field-induced release and MRI.

The synthesis of a novel nanocapsule-based carrier system is described, possessing a triggered release in remote-controlled fashion upon application of an external magnetic field in combination with the possibility to use the capsules as contrast agents for magnetic resonance imaging (MRI). Therefore, polymeric nanocontainers containing a high amount of superparamagnetic MnFe2 O4 nanoparticles and a thermo-degradable shell are fabricated via a miniemulsion route. The process allows the facile encapsulation of hydrophilic compounds, as demonstrated for a model dye. Release of the encapsulated dye is achieved upon application of an external alternating magnetic field. While the magnetic nanoparticles here act as heat generators to stimulate the decomposition of the shell and subsequently a release of the payload, they additionally enable the use of the nanocapsules as imaging agents for MRI. Due to the encapsulated magnetic nanoparticles, the nanocapsules possess high r2 relaxivity values of 96-120 Hz mmol(-1) , which makes them suitable for MRI. In toxicity experiments, the nanocapsules show no cell toxicity up to fairly high concentrations (600 µg mL(-1) ). Due to their dual-functionality, the nanocapsules possess high potential as nanocarriers with combined magnetic-field-induced release capability and as contrast agents for MRI.

Long-term active antimicrobial coatings for surgical sutures based on silver nanoparticles and hyperbranched polylysine

The goal of this study was to develop a long-term active antimicrobial coating for surgical sutures. To this end, two water-insoluble polymeric nanocontainers based on hyperbranched polylysine (HPL), hydrophobically modified by either using glycidyl hexadecyl ether, or a mixture of stearoyl/palmitoyl chloride, were synthesized. Highly stabilized silver nanoparticles (AgNPs, 2–5 nm in size) were generated by dissolving silver nitrate in the modified HPL solutions in toluene followed by reduction with L-ascorbic acid. Poly(glycolic acid)-based surgical sutures were dip-coated with the two different polymeric silver nanocomposites. The coated sutures showed high efficacies of more than 99.5% reduction of adhesion of living Staphylococcus aureus cells onto the surface compared to the uncoated specimen. Silver release experiments were performed on the HPL-AgNP modified sutures by washing them in phosphate buffered saline for a period of 30 days. These coatings showed a constant release of silver ions over more than 30 days. After this period of washing, the sutures retained their high efficacies against bacterial adhesion. Cytotoxicity tests using L929 mouse fibroblast cells showed that the materials are basically non-cytotoxic.

Enzymatic Cascade Reactions inside Polymeric Nanocontainers: A Means to Combat Oxidative Stress

Oxidative stress, which is primarily due to an imbalance in reactive oxygen species, such as superoxide radicals, peroxynitrite, or hydrogen peroxide, represents a significant initiator in pathological conditions that range from arthritis to cancer. Herein we introduce the concept of enzymatic cascade reactions inside polymeric nanocontainers as an effective means to detect and combat superoxide radicals. By simultaneously encapsulating a set of enzymes that act in tandem inside the cavities of polymeric nanovesicles and by reconstituting channel proteins in their membranes, an efficient catalytic system was formed, as demonstrated by fluorescence correlation spectroscopy and fluorescence cross-correlation spectroscopy. Superoxide dismutase and lactoperoxidase were selected as a model to highlight the combination of enzymes. These were shown to participate in sequential reactions in situ in the nanovesicle cavity, transforming superoxide radicals to molecular oxygen and water and, therefore, mimicking their natural behavior. A channel protein, outer membrane protein F, facilitated the diffusion of lactoperoxidase substrate/products and dramatically increased the penetration of superoxide radicals through the polymer membrane, as established by activity assays. The system remained active after uptake by THP-1 cells, thus behaving as an artificial organelle and exemplifying an effective approach to enzyme therapy.

Towards Targeted Drug Delivery by Covalent Ligand‐Modified Polymeric Nanocontainers

AbstractHere we present a novel strategy for specific cellular targeting of polymeric nanocontainers by using self‐assembly of block copolymers consisting of either Polydimethoxysiloxane‐b‐Polymethyloxazoline‐b‐Polydimethoxysiloxane (PDMS‐b‐PMOXA‐b‐PDMS) or functionalized PDMS‐b‐PMOXA‐b‐PDMS. Covalent functionalization of the above copolymer was accomplished using either the fluorescent dye sulforhodamine B or a poly‐guanosin ligand, the latter by using the Huisgen 1,3‐dipolar cycloaddition. The success of the covalent modification of the block copolymer has been determined by studying functionalized sulforhodamine B by NMR and fluorescence correlation spectroscopy. The covalent click chemistry approach leads to efficiently functionalized polymeric nanocontainers which enables specific uptake by activated macrophages overexpressing the scavenger receptor A1.

Branched poly- N-vinyl-2-pyrrolidones as polymeric nanocontainers for hydrophilic dyes

The capability of N-vinyl-2-pyrrolidone branched copolymers to incapsulate molecules of hydrophilic dyes has been found and studied.

Observing Proteins as Single Molecules Encapsulated in Surface‐Tethered Polymeric Nanocontainers

Immobilizing biomolecules provides the advantage of observing them individually for extended time periods, which is impossible to accomplish for freely diffusing molecules in solution. In order to immobilize individual protein molecules, we encapsulated them in polymeric vesicles made of amphiphilic triblock copolymers and tethered the vesicles to a cover slide surface. A major goal of this study is to investigate polymeric vesicles with respect to their suitability for protein-folding studies. The fact that polymeric vesicles possess an extreme stability under various chemical conditions is supported by our observation that harsh unfolding conditions do not perturb the structural integrity of the vesicles. Moreover, polymerosomes prove to be permeable to GdnHCl and, thereby, ideally suited for unfolding and refolding studies with encapsulated proteins. We demonstrate this with encapsulated phosphoglycerate kinase, which was fluorescently labeled with Atto655, a dye that exhibits pronounced photoinduced electron transfer (PET) to a nearby tryptophan residue in the native state. Under unfolding conditions, PET was reduced, and we monitored alternating unfolding and refolding conditions for individual encapsulated proteins.

Polymeric nanocontainers with high loading capacity of hydrophobic drugs

Water soluble 4- and 6-arm star-shaped polymers with (bio) degradable hydrophobic cores and dense hydrophilic coronas were synthesized by a combination of ring opening polymerization (ROP) and atom transfer radical polymerization (ATRP). Their carrier abilities of different model compounds as well as active pharmaceutically ingredients in aqueous solutions were evaluated by UV/Vis and H-1 NMR spectroscopy studies. The highest loading capacities (up to 36 guest molecules per molecule of polymer) were, as expected, found for the 6-arm star-shaped polymers. The densely grafted poly[poly(ethylene glycol) methyl ether methacrylate] [p(mPEGMA)] shell stabilizes the host-guest complexes preventing undesired multimolecular aggregation. Dynamic light scattering (DLS) measurements evidenced the unimolecular character of all 4- and 6-arm polymers before and after loading with guest molecules. The (bio) degradability of the core was demonstrated under acidic conditions and in vitro using a lipase from Rhizopus Arrhizus. The dual-stimuli responsiveness of the hydrophobic core to enzymes and pH may facilitate increased control over in vivo drug release.

A novel use of an in vitro method to predict the in vivo stability of block copolymer based nano-containers

The purpose of this study was to design an in vitro experiment that can assess the stability of polymeric micellar formulations of hydrophobic drugs such as cyclosporine A (CyA) in blood, and predict the in vivo performance of the examined delivery system. Poly(ethylene oxide)- block-poly(ε-caprolactone) (PEO- b-PCL) copolymers were assembled to polymeric nano-containers for the physical encapsulation of CyA by a co-solvent evaporation method using different loading conditions. CyA-loaded micelles were prepared and compared to commercially available intravenous formulation of CyA (Sandimmune®) for in vitro release, protein binding, and pharmacokinetic parameters in Sprague–Dawley rats. The unbound fraction ( fu) of CyA was determined using an erythrocyte vs. plasma and buffer partitioning technique. Different polymeric micellar formulations of CyA did not show any significant difference in CyA release when dialyzed against bovine serum albumin. The fu experiments, however, revealed a significant decrease in the fu of the loaded drug with an increase in the drug/polymer loading ratio, while the fu of all micellar formulations were significantly lower than Sandimmune®. The pharmacokinetic study showed that fu of CyA in each formulation correlated with its in vivo performance determined by pharmacokinetic parameters: the lower fu of the formulation, translated to a higher area under the concentration versus time curve (AUC), and a lower clearance (CL) and volume of distribution (Vd). In conclusion, determination of the unbound fraction of encapsulated drug can be used to predict the in vivo stability of polymeric micellar nano-containers. PEO- b-PCL micelles containing higher CyA-loaded levels are shown to be more stable changing the pharmacokinetics of the encapsulated CyA to a higher extent.

Encapsulation of Fluorescent Molecules by Functionalized Polymeric Nanocontainers: Investigation by Confocal Fluorescence Imaging and Fluorescence Correlation Spectroscopy

Nanocontainers (NCs) were prepared from amphiphilic triblock copolymers, having an average molecular weight of around 8000 g/mol, by using previously published preparation methods consisting of dispersing the polymer in an aqueous buffer solution containing molecules for encapsulation. A small molecular weight fluorophore, sulforhodamine B, as well as the fluorescent protein avidin labeled with Alexa 488 were encapsulated, and the resulting nanocontainers were characterized using fluorescence correlation spectroscopy (FCS) and fluorescence cross-correlation spectroscopy (FCCS). Nanocontainer size determination by FCS is very robust and compares well with results obtained from photon correlation spectroscopy: the measured diameters of the polymeric nanocontainers vary between 140 and 172 nm. Encapsulation of fluorescent molecules was determined by evaluating the molecular brightness of nanocontainers with an encapsulated fluorescently labeled protein (avidin-Alexa 488). Results indicate that the number of encapsulated avidin-Alexa 488 molecules corresponds well with the initial concentration of the fluorescently labeled protein and the encapsulated volume. A nanocontainer binding assay was developed using biotinylated fluorescently labeled nanocontainers. Binding of biotinylated nanocontainers to fluorescently labeled streptavidin was followed by fluorescence cross-correlation spectroscopy. The intrinsic dissociation constant, K(d), of labeled streptavidin to the ligand-modified nanocontainers is 1.7 +/- 0.4 x 10(-8) M, and about 1921 +/- 357 molecules of labeled streptavidin are bound to each nanocontainer.