Muharrem Seleci

Nanostructured Amphiphilic Star-Hyperbranched Block Copolymers for Drug Delivery

A robust drug delivery system based on nanosized amphiphilic star-hyperbranched block copolymer, namely, poly(methyl methacrylate-block-poly(hydroxylethyl methacrylate) (PMMA-b-PHEMA) is described. PMMA-b-PHEMA was prepared by sequential visible light induced self-condensing vinyl polymerization (SCVP) and conventional vinyl polymerization. All of the synthesis and characterization details of the conjugates are reported. To accomplish tumor cell targeting property, initially cell-targeting (arginylglycylaspactic acid; RGD) and penetrating peptides (Cys-TAT) were binding to each other via the well-known EDC/NHS chemistry. Then, the resulting peptide was further incorporated to the surface of the amphiphilic hyperbranched copolymer via a coupling reaction between the thiol (-SH) group of the peptide and the hydroxyl group of copolymer by using N-(p-maleinimidophenyl) isocyanate as a heterolinker. The drug release property and targeting effect of the anticancer drug (doxorobucin; DOX) loaded nanostructures to two different cell lines were evaluated in vitro. U87 and MCF-7 were chosen as integrin αvβ3 receptor positive and negative cells for the comparison of the targeting efficiency, respectively. The data showed that drug-loaded copolymers exhibited enhanced cell inhibition toward U87 cells in compared to MCF-7 cells because targeting increased the cytotoxicity of drug-loaded copolymers against integrin αvβ3 receptor expressing tumor cells.

Amine-intercalated montmorillonite matrices for enzyme immobilization and biosensing applications

Clay based biosensors were developed using montmorillonite (Mont) modified with methyl (M) and dimethylamine (DM). X-ray diffraction, Fourier transform infrared spectroscopy, zeta potential and thermal gravimetric measurements were used to characterize the modified clays. After immobilization of glucose oxidase (GOx) via clay on the glassy carbon electrode, its application as a glucose biosensor was investigated in detail. The best response characteristics were obtained by DM-Mont and optimization of enzyme amount, reproducibility of biosensor fabrication, repeatability of measurements and operational stability were all evaluated. The optimized biosensor showed a very good linearity between 0.05 mM and 1.0 mM, a 7 s response time and a limit of detection to glucose of 0.038 mM. Also, kinetic parameters and stabilities were determined. Apparent Km and Imax values were found as 0.73 mM and 2.955 μA, respectively. As well as batch configuration, the DM-Mont/GOx biosensor was successfully applied in the flow injection analysis mode. Finally, the performance of the DM-Mont/GOx biosensor to analyze glucose in a wine sample was compared with HPLC.

Biofunctional quantum dots as fluorescence probe for cell-specific targeting☆

We describe here the synthesis, characterization, bioconjugation, and application of water-soluble thioglycolic acid TGA-capped CdTe/CdS quantum dots (TGA-QDs) for targeted cellular imaging. Anti-human epidermal growth factor receptor 2 (HER2) antibodies were conjugated to TGA-QDs to target HER2-overexpressing cancer cells. TGA-QDs and TGA-QDs/anti-HER2 bioconjugates were characterized by fluorescence and UV-Vis spectroscopy, X-ray diffraction (XRD), hydrodynamic sizing, electron microscopy, and gel electrophoresis. TGA-QDs and TGA-QDs/anti-HER2 were incubated with cells to examine cytotoxicity, targeting efficiency, and cellular localization. The cytotoxicity of particles was measured using an MTT assay and the no observable adverse effect concentration (NOAEC), 50% inhibitory concentration (IC50), and total lethal concentration (TLC) were calculated. To evaluate localization and targeting efficiency of TGA-QDs with or without antibodies, fluorescence microscopy and flow cytometry were performed. Our results indicate that antibody-conjugated TGA-QDs are well-suited for targeted cellular imaging studies.

Folic acid-modified clay: targeted surface design for cell culture applications

Here we report the preparation, characterization and application of folic acid modified Montmorillonite clay (FA-Mont) as a cell culture material. Clays exhibit unique properties such as good mechanical and chemical stabilities, high surface area, low toxicity and easy combination with functional organic groups. FA was used as a modifier to facilitate adhesion of folate positive cells on the clay surface. FA-Mont was characterized using FT-IR, XRD, zeta potential measurements as well as thermogravimetric analyses. Finally, the usage potential of FA-Mont as a receptor mediated cell adhesion material was successfully proved by using folate receptor (FR) rich HeLa and FR poor A-549 cells. The cell adhesion studies were monitored and imaged by fluorescence and scanning electron microscopy techniques.

Amine intercalated clay surfaces for microbial cell immobilization and biosensing applications

Trimethylamine (TM) intercalated montmorillonite (Mont) clay was prepared and characterized using X-ray diffraction, Fourier transform infrared spectroscopy, zeta potential and thermal gravimetric measurements. Gluconobacter oxydans cells were immobilized on the micro-structured matrix and used in microbial sensor applications. Sensor responses were based on the respiratory activity of the cells and the consumption of oxygen was monitored at −0.7 V (vs. Ag/AgCl and platinum electrodes) by using glucose as a substrate. Stabilization of the bacteria was performed on the Mont using glutaraldehyde. Optimization of the TM-Mont/G. oxydans sensor and examination of the electrochemical mechanism were carried out in a batch system. Measurements concerning analytical characteristics, operational stability, repeatability and substrate specificity depending on the carbon source in the culture were investigated in a flow injection analysis (FIA) system. Linear ranges were found between 0.15 and 5.0 mM for the batch mode and 0.1 and 5.0 mM for the FIA system, respectively. Finally, real samples were analyzed and were compared with the results of a spectrophotometric method as reference.

From Invisible Structures of SWCNTs toward Fluorescent and Targeting Architectures for Cell Imaging

Single-walled carbon nanotubes (SWNTs) are unique nanostructures used as cargo systems for variety of diagnostic and therapeutic agents. For taking advantage of these structures in biological processes, they should be visible. Therefore, fluorescence labeling of SWCNTs with various probes is a significant issue. Herein, we demonstrate a simple approach for cell specific imaging and diagnosis by combining SWCNTs with a copolymer poly(para-phenylene) (PPP) containing polystyrene (PSt) and poly(ε-caprolactone) (PCL) side chains (PPP-g-PSt-PCL). In this approach PPP-g-PSt-PCL is noncovalently attached on carboxyl functional SWCNTs. The obtained fluorescent probe is bound to folic acid (FA) for targeted imaging of folate receptor (FR) positive HeLa cells. In vitro studies demonstrate that this conjugate can specifically bind to HeLa cells and indicate great potential for targeting and imaging studies.

Niosomes as Nanoparticular Drug Carriers

Drug delivery systems are defined as formulations aiming for transportation of a drug to the desired area of action within the body. The basic component of drug delivery systems is an appropriate carrier that protects the drug from rapid degradation or clearance and thereby enhances drug concentration in target tissues. Based on their biodegradable, biocompatible, and nonimmunogenic structure, niosomes are promising drug carriers that are formed by self-association of nonionic surfactants and cholesterol in an aqueous phase. In recent years, numerous research articles have been published in scientific journals reporting the potential of niosomes to serve as a carrier for the delivery of different types of drugs. The present review describes preparation methods, characterization techniques, and recent studies on niosomal drug delivery systems and also gives up to date information regarding recent applications of niosomes in drug delivery.

Fluorescent micellar nanoparticles by self-assembly of amphiphilic, nonionic and water self-dispersible polythiophenes with “hairy rod” architecture

Polymers with “hairy-rod” architecture having oligo/polythiophene (PTh) as main chain and poly (ethylene glycol) (PEG) (Mn = 2000) as flexible side chains were obtained by combining the “macromonomer technique” with specific methods for the synthesis of conjugated polymers. Fluorescent nanoparticles of core–shell type with high colloidal stability were obtained from these water self-dispersible materials by a direct dissolution method in aqueous media. It has been shown that the size and photophysical properties of the micellar nanoparticles in aqueous dispersions as well as the bulk properties of the investigated materials can be tuned by varying the PEG side chain density and by the modality of PEG connection to the PTh main chains. The presence of the PEG shells in the structure of these fluorescent nanoparticles cans suppress the non-specific interactions with biomolecules on the one side and on the other side they work as a biomimetic interface that could facilitate their potential use as cell-imaging agents. The present attempt offers an ease of access alternative to conducting polymer nanoparticle encapsulation in a biocompatible matrix by nanoprecipitation.

Aptamer mediated niosomal drug delivery

Development of nanoscale carrier systems for targeted drug delivery is crucial for cancer treatment. The current methods of drug delivery exhibit some problems such as lack of therapy efficiency at the desired parts of the body, degradation of the drug before reaching the desired tissue and limitations in cellular penetration. In this work, a novel drug delivery platform was developed to overcome these problems and to enable specific and efficient uptake into the cells. The surface of the synthesized polyethylene glycolated niosomes (PEGNIO) was modified with cell penetrating peptide (CPP) and cell specific MUC1 (S2.2) aptamer, and doxorubicin (DOX) as a cancer model drug was encapsulated in this platform. Fluorescence microscopy and flow cytometry analysis were used to investigate the cellular uptake and intracellular distribution of the DOX loaded niosomal formulation. In vitro cytotoxicity studies were carried out using MUC1 positive HeLa and negative U87 cells. Moreover, dynamic light scattering (DLS), zeta potential measurements and fluorescence absorption spectroscopy were performed to determine the vesicle size, as well as charge and spectroscopic properties of the conjugates. From these results, this novel aptamer mediated niosomal drug delivery platform may have application potential in targeted drug delivery towards MUC1-overexpressing tumors.

Phyto-Niosomes: In Vitro Assessment of the Novel Nanovesicles Containing Marigold Extract

Herbal compounds, so-called phytoconstituents, illustrate poor absorption by living cells. Phytosomes are advanced form of herbal compounds that show higher absorption rate and bioavailability, which results better than conventional plant extracts. Niosomes, which are made of nonionic surfactants, create better chemical and stability conditions besides lipid vesicles. This study covers the preparation, characterization and cell culture applications of phyto-niosomes of Marigold extract. Before the encapsulation process, extracts of selected plants were prepared and the extract that presents best bioactivity was chosen. The resulting phyto-niosomes were characterized and their biological activities including cytotoxicity, wound healing and antioxidant activity were investigated. GRAPHICAL ABSTRACT