The EcCLC antiporter embedded in lipidic liquid crystalline films - molecular dynamics simulations and electrochemical methods.

Lipidic-liquid crystalline nanostructures (lipidic cubic phases), which are biomimetic and stable in an excess of water, were used as a convenient environment to investigate the transport properties of the membrane antiporter E. coli CLC-1 (EcCLC). The chloride ion transfer by EcCLC was studied by all-atom molecular dynamics simulations combined with electrochemical methods at pH 7 and pH 5. The cubic phase film was used as the membrane between the chloride donor and receiving compartments and it was placed on the glassy carbon electrode and immersed in the chloride solution. Structural characterization of lipidic mesoscopic systems with and without the incorporation of EcCLC was performed using small-angle X-ray scattering. The EcCLC transported chloride ions more efficiently at more acidic pH, and the resistance of the film decreased at lower pH. 4,4-Diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) employed as an inhibitor of the protein was shown to decrease the transport efficiency upon hydrolysis to DADS at both pH 7 and pH 5. The molecular dynamics simulations, performed for the first time in lipidic cubic phases for EcCLC, allowed studying the collective movements of chloride ions which can help in elucidating the mechanism of transporting the ions by the EcCLC antiporter. The protein modified lipidic cubic phase film is a convenient and simple system for screening potential inhibitors of integral membrane proteins, as demonstrated by the example of the EcCLC antiporter. The use of lipidic cubic phases may also be important for the further development of new electrochemical sensors for membrane proteins and enzyme electrodes.

Interactions of doxorubicin with organized interfacial assemblies. 1. Electrochemical characterization.

Doxorubicin is an anthracycline that has found wide use as a chemotherapeutic agent, with the primary target of its action being nuclear DNA. Despite the large body of knowledge on this family of compounds, the mechanism of doxorubicin penetration through the cellular or nuclear membrane remains understood to a limited extent. The plasma membrane acts as a barrier to the permeation of polar molecules, and this effect is mainly due to the hydrophobicity of membrane interior. The partitioning of DOX molecules into the lipid bilayer must thus be the basis for its passive transport across the biological membrane and therefore a key area of research activity lies in understanding how the structure of the anthracycline influences its interactions with amphiphilic interfaces. We have studied interactions between doxorubicin and Langmuir/Langmuir-Blodgett monomolecular films of octadecylamine (C18NH2), dihexadecylphosphate (DHP) and 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), and DMPC bilayer films (Langmuir-Schaeffer) on a polycrystalline gold surface using ellipsometry, cyclic voltammetry, electrochemical impedance spectroscopy, and quartz crystal microbalance measurements. For all biomimetic films there is a substantial interaction between doxorubicin and the interface, and the extent of this interaction depends on the hydrophobic/hydrophilic properties of the film formed and its organization.

Interactions of doxorubicin with organized interfacial assemblies. 2. Spectroscopic characterization.

Doxorubicin is an anthracycline that has found wide use as a chemotherapeutic agent, with the primary limitation to its use being cardiotoxicity. Depending on the identity and location of pendent side groups, the anthracyclines exhibit varying degrees of chemotherapeutic activity and toxicity, and a key area of research activity lies in understanding how the structure of the anthracycline influences its interactions with amphiphilic interfaces. We have studied interactions between doxorubicin and interfacial adlayers of octadecylamine (C18NH2), dihexadecylphosphate (DHP), and both monolayers and bilayers of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) on mica using time- and frequency-resolved spectroscopic measurements. We report surface-enhanced resonance Raman data and fluorescence lifetime and anisotropy imaging data for doxorubicin at these interfaces. For all monolayers, there is a substantial interaction between doxorubicin and the interface. For DMPC bilayers, the extent of the interaction between doxorubicin and the interface depends on how the interface was formed.

Synthesis and Characterization of Magnetic Drug Carriers Modified with Tb3+ Ions.

The study aimed to synthesize and characterize the magnetic drug carrier modified with terbium (III) ions. The addition of terbium extends the possibilities of their applications for targeted anticancer radiotherapy as well as for imaging techniques using radioisotopes emitting ß+, ß-, α, and γ radiation. The synthesis of iron oxide nanoparticles stabilized with citrates using the co-precipitation method (IONP @ CA) was carried out during the experimental work. The obtained nanoparticles were used to synthesize a conjugate containing terbium ions and guanosine-5'-monophosphate as an analog of drugs from the thiopurine group. Conjugates and their components were characterized using Transmission Electron Microscopy, infrared spectroscopy, X-ray microanalysis, spectrofluorimetry, and thermogravimetric analysis. The hybrid was also investigated with Langmuir layers to check the interaction with analogs of biological membranes.

Interactions of doxorubicin with self-assembled monolayer-modified electrodes: electrochemical, surface plasmon resonance (SPR), and gravimetric studies.

We present the results on the partitioning of doxorubicin (DOX), a potent anticancer drug, through the model membrane system, self-assembled monolayers (SAMs) on gold electrodes. The monolayers were formed from alkanethiols of comparable length with different ω-terminal groups facing the aqueous electrolyte: the hydrophobic -CH(3) groups for the case of dodecanethiol SAMs or hydrophilic -OH groups of mercaptoundecanol SAMs. The electrochemical experiments combined with the surface plasmon resonance (SPR) and gravimetric studies show that doxorubicin is likely adsorbed onto the surface of hydrophilic monolayer, while for the case of the hydrophobic one the drug mostly penetrates the monolayer moiety. The adsorption of the drug hinders further penetration of doxorubicin into the monolayer moiety.

Hybrid System for Local Drug Delivery and Magnetic Hyperthermia Based on SPIONs Loaded with Doxorubicin and Epirubicin.

Cancer is one of the most common causes of death worldwide, thus new solutions in anticancer therapies are highly sought after. In this work, superparamagnetic iron oxide nanoparticles (SPIONs) conjugated with anticancer drugs are synthesized and investigated as potential magnetic drug nanocarriers for local drug delivery and mild magnetic hyperthermia. We have obtained a hybrid system loaded with holmium and anticancer drugs and thoroughly studied it with respect to the size, morphology, surface modifications and magnetic properties, and interactions with the model of biological membranes, cytotoxicity. We present that nanoparticles having a round shape and size 15 nm are successfully stabilized to avoid their agglomeration and modified with doxorubicin or epirubicin within a controlled way. The number of drugs loaded into the SPIONs was confirmed with thermogravimetry. The hybrid based on SPIONs was investigated in touch with model biological membranes within the Langmuir-Blodgett technique, and results show that modified SPION interacts effectively with them. Results obtained with magnetic hyperthermia and biological studies confirm the promising properties of the hybrid towards future cancer cell treatment.

Physicochemical properties and in vitro cytotoxicity of iron oxide-based nanoparticles modified with antiangiogenic and antitumor peptide A7R.

Superparamagnetic iron oxide-based nanoparticles (SPIONs) are promising carriers as targeted drug delivery vehicles, because they can be guided to their target with the help of an external magnetic field. Functionalization of nanoparticles' surface with molecules, which bind with high affinity to receptors on target tissue significantly facilitates delivery of coated nanoparticles to their targeted site. Here, we demonstrate conjugation of an antiangiogenic and antitumor peptide ATWLPPR (A7R) to SPIONs modified with sebacic acid (SPIONs-SA). Successful conjugation was confirmed by various analytical techniques (FTIR, SERS, SEM-EDS, TEM, TGA). Cell cytotoxicity studies, against two cell lines (HUVEC and MDA-MB-231) indicated that SPIONs modified with A7R reduced HUVEC cell viability at concentrations higher than 0.01 mg Fe/mL, in comparison to cells that were exposed to either the nanoparticles modified with sebacic acid or A7R peptide solely, what might be partially caused by a process of internalization.

Monoolein Cubic Phase Gels and Cubosomes Doped with Magnetic Nanoparticles-Hybrid Materials for Controlled Drug Release.

Hybrid materials consisting of a monoolein lipidic cubic phase (LCP) incorporating two types of magnetic nanoparticles (NP) were designed as addressable drug delivery systems. The materials, prepared in the form of a gel, were subsequently used as a macroscopic layer modifying an electrode and, after dispersion to nanoscale, as magnetocubosomes. These two LCPs were characterized by small-angle X-ray scattering (SAXS), cross-polarized microscopy, magnetic measurements, and phase diagrams. The magnetic dopants were hydrophobic NPoleic and hydrophilic NPcitric, characterized by dynamic light scattering (DLS) and transmission electron microscopy (TEM), and their influence on the properties of the cubic phases was investigated. The removal of the anticancer drug, Doxorubicin (Dox) from the hybrid cubic phase gels was studied by electrochemical methods. The advantages of incorporating magnetic nanoparticles into the self-assembled lipid liquid crystalline phases include the ability to address the cubic phase nanoparticle containing large amounts of drug and to control the kinetics of the drug release.

Solid-core and hollow magnetic nanostructures: Synthesis, surface modifications and biological applications.

In the need of development of versatile and flexible platforms for sensing, nanostructured particles are one of the systems of choice. Additionally, the state-of-the-art, controlled surface modifications of these structures offer broad possibilities of using such systems for diagnostics and therapy, often referred to as thera(g)nostics. In this brief review we will focus on the synthesis and surface modifications of solid-core magnetic nanostructures and polymeric capsules containing nanoferrites modified with anti-cancer drug--doxorubicin, designed for magnetic field-driven drug delivery for cancer therapy. We will also outline some problems related to the usage of such structures. The encapsulation and distribution of magnetic iron oxide nanoparticles modified with doxorubicin will be demonstrated in the polypyrrole spherical microvessels.