Sacide Alsoy Altinkaya

Effect of PEG Grafting Density and Hydrodynamic Volume on Gold Nanoparticle–Cell Interactions: An Investigation on Cell Cycle, Apoptosis, and DNA Damage

In this study, interactions of polyethylene glycol (PEG)-coated gold nanoparticles (AuNPs) with cells were investigated with particular focus on the relationship between the PEG layer properties (conformation, grafting density, and hydrodynamic volume) and cell cycle arrest, apoptosis, and DNA damage. Steric hindrance and PEG hydrodynamic volume controlled the protein adsorption, whereas the AuNP core size and PEG hydrodynamic volume were primary factors for cell uptake and viability. At all PEG grafting densities, the particles caused significant cell cycle arrest and DNA damage against CaCo2 and PC3 cells without apoptosis. However, at a particular PEG grafting density (∼0.65 chains/nm(2)), none of these severe damages were observed on 3T3 cells indicating discriminating behavior of the healthy (3T3) and cancer (PC3 and CaCo2) cells. It was concluded that the PEG grafting density and hydrodynamic volume, tuned with the PEG concentration and AuNP size, played an important role in particle-cell interactions.

Surface modification of polysulfone based hemodialysis membranes with layer by layer self assembly of polyethyleneimine/alginate-heparin: a simple polyelectrolyte blend approach for heparin immobilization

This study intends to improve blood compatibility of polysulfone (PSF) membranes by generating a nonthrombogenic surface through heparin immobilization. To achieve this task, the support membrane prepared from a blend of PSF and sulfonated polysulfone (SPSF) was modified with layer by layer (LBL) deposition of polyethyleneimine (PEI) and alginate (ALG) and heparin blended with ALG was immobilized only on the outermost surface of the LBL assembly. The results have shown that the adsorption of human plasma proteins and platelet activation on the LBL modified membranes decreased significantly compared with the unmodified PSF and PSF-SPSF blend membranes. Furthermore, blending ALG with a small amount of heparin remarkably prolonged the APTT values of heparin free PEI/ALG coated membranes. It is envisaged that the use of a blend of HEP and ALG only in the terminating layer of the LBL assembly can be an economical and alternative modification technique to create nonthrombogenic surfaces.

Multi-Zone Drying Schemes for Lowering the Residual Solvent Content during Multi-Component Drying of Semicrystalline Polymers

The development of a glassy skin in multicomponent semicrystalline polymer systems limits the diffusion of solvents out of the system and increases residual solvent levels. Based on the results of a mathematical model that we had previously developed, we have proposed a multi-zone drying scheme aimed at lowering the residual solvent levels by taking into account the effect of interactions between the various solvents as predicted by the model. This article focuses on the application of this model to develop optimal drying schemes and to verify the effectiveness of these predictions using experimental techniques. The mathematical model developed previously to study the diffusion of multiple solvents and changes in the crystallinity of semicrystalline polymer systems during drying incorporates many features including Vrentas-Duda diffusion theory, solvent-induced crystallization kinetics, as well as glass transition effects and skinning of the film. The multi-zone drying system was developed by varying the drying temperature in each zone as well as changing the partial pressure of individual solvents during the drying process. The effectiveness of the multi-zone drying schemes predicted by the model was validated experimentally using thermogravimetric methods. The polymer-solvent system chosen was a poly(vinyl alcohol)-water-methanol system. Our experimental data suggested that the multi-zone drying schemes were superior to a single-zone drying system through direct comparison. Further examination of the mathematical model yielded individual solvent profiles and these data reaffirmed our conclusions that a multi-zone drying scheme has the ability to reduce the effect of solvent trapping and thus lower the overall residual solvent content.

Gelatin-based 3D conduits for transdifferentiation of mesenchymal stem cells into Schwann cell-like phenotypes

In this study, gelatin-based 3D conduits with three different microstructures (nanofibrous, macroporous and ladder-like) were fabricated for the first time via combined molding and thermally induced phase separation (TIPS) technique for peripheral nerve regeneration. The effects of conduit microstructure and mechanical properties on the transdifferentiation of bone marrow-derived mesenchymal stem cells (MSCs) into Schwann cell (SC) like phenotypes were examined to help facilitate neuroregeneration and understand material-cell interfaces. Results indicated that 3D macroporous and ladder-like structures enhanced MSC attachment, proliferation and spreading, creating interconnected cellular networks with large numbers of viable cells compared to nanofibrous and 2D-tissue culture plate counterparts. 3D-ladder-like conduit structure with complex modulus of ∼0.4×106Pa and pore size of ∼150μm provided the most favorable microenvironment for MSC transdifferentiation leading to ∼85% immunolabeling of all SC markers. On the other hand, the macroporous conduits with complex modulus of ∼4×106Pa and pore size of ∼100μm showed slightly lower (∼65% for p75, ∼75% for S100 and ∼85% for S100β markers) immunolabeling. Transdifferentiated MSCs within 3D-ladder-like conduits secreted significant amounts (∼2.5pg/mL NGF and ∼0.7pg/mL GDNF per cell) of neurotrophic factors, while MSCs in macroporous conduits released slightly lower (∼1.5pg/mL NGF and 0.7pg/mL GDNF per cell) levels. PC12 cells displayed enhanced neurite outgrowth in media conditioned by conduits with transdifferentiated MSCs. Overall, conduits with macroporous and ladder-like 3D structures are promising platforms in transdifferentiation of MSCs for neuroregeneration and should be further tested in vivo. STATEMENT OF SIGNIFICANCE This manuscript focuses on the effect of microstructure and mechanical properties of gelatin-based 3D conduits on the transdifferentiation of mesenchymal stem cells to Schwann cell-like phenotypes. This work builds on our recently accepted manuscript in Acta Biomaterialia focused on multifunctional 2D films, and focuses on 3D microstructured conduits designed to overcome limitations of current strategies to facilitate peripheral nerve regeneration. The comparison between conduits fabricated with nanofibrous, macroporous and ladder-like microstructures showed that the ladder-like conduits showed the most favorable environment for MSC transdifferentiation to Schwann-cell like phenotypes, as seen by both immunolabeling as well as secretion of neurotrophic factors. This work demonstrates the importance of controlling the 3D microstructure to facilitate tissue engineering strategies involving stem cells that can serve as promising approaches for peripheral nerve regeneration.

Responsive pentablock copolymers for siRNA delivery

In this study, temperature and pH responsive cationic and amphiphilic pentablock copolymers, which consist of the temperature responsive triblock Pluronic F127 sandwiched between pH responsive PDEAEM (poly(2-diethylaminoethyl methacrylate)) end blocks, were used for the first time in the development of polyplex and gold nanoparticle (AuNP) based multicomponent siRNA delivery systems (MCSs). Copolymers in both systems protected siRNA from external effects, provided cell entry and endosomal escape. The thermoreversible micellization of the hydrophobic PPO block facilitated the cellular entry while the PDEAEM blocks enhanced the endosomal escape through protonated tertiary amine groups by pH buffering. The synergistic advantages of the different blocks showed an enhanced effect in the MCSs due to attachment and surface configuration reasons. The siRNA transfection efficiency of MCSs against luciferase expressing SKOV3 cells was 15% higher than both the polyplexes alone and the commercial siRNA transfection agent Lipofectamine RNAiMax at the same applied dose, without any toxicity. The results indicated that the multicomponent systems based on the responsive cationic pentablock copolymers and gold nanoparticles have promising potential as an efficient siRNA delivery vector for future applications.

Nanofibrous gelatine scaffolds integrated with nerve growth factor-loaded alginate microspheres for brain tissue engineering

Neural regeneration research is designed in part to develop strategies for therapy after nerve damage due to injury or disease. In this study, a new gelatine‐based biomimetic scaffold was fabricated for brain tissue engineering applications. A technique combining thermally induced phase separation and porogen leaching was used to create interconnected macropores and nanofibrous structure. To promote tissue regeneration processes, the scaffolds were integrated with nerve growth factor (NGF)‐loaded alginate microspheres. The results showed that nanofibrous matrix could only be obtained when gelatine concentration was at least 7.5% (w/v). The scaffold with a modulus value (1.2 kPa) similar to that of brain tissue (0.5–1 kPa) was obtained by optimizing the heat treatment time, macropore size and gelatine concentration. The encapsulation efficiencies of NGF into 0.1% and 1% alginate microspheres were 85% and 100%, respectively. The release rate of NGF from the microspheres was controlled by the alginate concentration and the poly(L‐lysine) coating. The immobilization of the microspheres in the scaffold reduced burst release and significantly extended the release period. The nanofibrous architecture and controlled release of NGF from the microspheres induced neurite extension of PC12 cells, demonstrating that the released NGF was in an active form. The results suggest that the scaffolds prepared in this study may have potential applications in brain tissue engineering due to topologic and mechanical properties similar to brain tissue and pore structure suitable for cell growth and differentiation. Copyright

The effects of urease immobilization on the transport characteristics and protein adsorption capacity of cellulose acetate based hemodialysis membranes

In this study, cellulose acetate (CA) based hemodialysis membranes were prepared by a dry phase inversion method and the influences of urease immobilization on the clearing performance and protein adsorption capacity of the membranes were investigated. Permeation experiments have shown that modification of CA membranes with urease immobilization not only enhanced the transport rate of urea but also increased the permeation coefficients of uric acid and creatinine by changing the structure of the membrane. Furthermore, the protein adsorption capacity of the CA membranes decreased. On the other hand, the mechanical strength of the modified CA membrane did not change significantly compared with that of the unmodified one. A mathematical model was derived to determine the rate of mass transfer of urea through modified CA membranes. Model predictions along with the experimental data suggest that urease immobilization can be used as an alternative method in preparing CA based hemodialysis membranes with improved transport characteristics and biocompatibility through reduced protein adsorption capacities.

Modeling of Hemodialysis Operation

In this study, a theoretical model was developed to predict the solute concentrations in patients’ blood and optimize the efficiency of the hemodialysis operation. The model takes into account simultaneous mass and momentum transfer on the blood side both in radial and axial directions. A key component of the model is the incorporation of the protein adsorption on the inner surface of the membrane. The validity of the model was confirmed with the experimental data available in the literature for two different types of hemodiafilter. To illustrate the importance of including the radial concentration gradients and protein adsorption kinetics in the model, the experimental data were predicted with and without consideration of these effects. The results have shown that assuming uniform concentration in the radial direction or neglecting protein adsorption on the inner surface of the membrane leads to higher error in predicting the experimental data. In addition, significant error can be introduced in the calculation of the dialysis time if protein adsorption is not considered.