Chemical Engineering / Kimya Mühendisliği
Permanent URI for this collectionhttps://hdl.handle.net/11147/14
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Research Project Kontrollü salım özelliklerine sahip antimikrobiyel, nanokompozit gıda ambalaj filmlerinin geliştirilmesi(TÜBİTAK - Türkiye Bilimsel ve Teknolojik Araştırma Kurumu, 2013) Alsoy Altınkaya, Sacide; Pekşen Özer, Bahar Başak; Oymacı, Pelin; Uz, Metin; Büyüköz, MeldaAntimikrobiyal gıda ambalaj malzemelerinde aktif maddenin kontrollü salımı kritiktir. Bu çalışmada kontrollü salım antimikrobiyal ajanı nanotanecik içine yükleyerek sağlanmıştır. Doğal bir antimikrobiyal ajan olan lisozim serbest halde ya da poliakrilik asit (PAA) ile tanecik oluşturarak peynir altı suyu izolatı (WPI) filmlerine ilave edilmiştir. Serbest lisozim içeren filmlerden salımın çok hızlı olduğu, 45 dakika gibi kısa bir sürede lisozimin tamamının ortama geçtiği, PAA içine yüklenmesi durumunda ise salım hızının oldukça yavaşladığı gözlenmiştir. PAA’nın molekül ağırlığı ile PAA/LIS oranının lisozimin salım hızı üzerinde oldukça etkili olduğu belirlenmiştir. Her iki parametrenin artması ile daha büyük boyutta tanecikler elde edilmiş ve bu tanecikleri içeren filmlerden lisozimin salım hızı düşmüştür. Film hazırlama esnasında lisozimin aktivitesinde kayıp yaşanmış, ancak salınan aktivite zamanla değişim göstermemiştir. Lisozim hem serbest, hem de PAA/LIS kompleksi içinde filme ilave edildiğinde ortama kontrollü bir şekilde salınarak antimikrobiyal etkisini daha uzun süre koruyabilmiştir. Çalışmada PAA/LIS komplekslerinin yanısıra kitosan nanotanecikler de hazırlanmış, bu taneciklerin WPI filmlerinin mekanik ve bariyer özellikleri üzerine olan etkileri incelenmiştir. %2 oranına kadar kitosan nanotanecik içeren filmlerin mekanik ve bariyer özelliklerinde önemli bir değişiklik olmazken, % 4 nanotanecik ilavesiyle çekme mukavemetinde ani bir artış, % uzama değerinde ve su buharı geçirgenliğinde de azalma gözlenmiştir. Ayrıca, kitosan nanotanecikler WPI filmlere antibakteriyel özellik de kazandırmıştır. Filme ilave edilen nanotanecik miktarının % 6’ya artırılması sonucunda homojen dağılımın sağlanamamasından dolayı mekanik ve bariyer özelliklerindeki iyileşme azalmaya başlamıştır. Bu çalışmada elde edilen sonuçlara dayanarak kitosan nanotaneciklerin WPI filminin mekanik ve bariyer özelliklerinin iyileştirilmesinde, PAA/LIS komplekslerinin de kontrollü salım yapabilen, antimikrobiyal özelliğe sahip WPI filmlerinin hazırlanmasında kullanılabileceği düşünülmektedir.Conference Object Development of Functional Materials for Sirna Delivery and Neural Tissue Engineering(AIChE, 2015) Uz, Metin; Alsoy Altınkaya, Sacide; Mallapragada, Surya K.The current nonviral siRNA delivery systems in the literature face many problems such as, cellular entry, endosomal escape and efficient siRNA release. Considering this motive, we developed gold nanoparticles (AuNPs) and temperature/pH responsive pentablock copolymer based siRNA delivery systems to address these problems. The temperature and pH responsive cationic and amphiphilic pentablock copolymers, which were consisted of the temperature responsive Pluronic F127 middle block constructed by PEO-PPO-PEO ((poly(ethyleneoxide)-block-poly(propyleneoxide)-block-poly(ethyleneoxide))) blocks contributing cellular entry through temperature responsive micellization and pH responsive cationic PDEAEM (poly(2-diethylaminoethyl methacrylate)) end blocks facilitating nucleic acid condensation and endosomal escape, were used for the first time in the development of polyplex and AuNP based multicomponent siRNA delivery systems (MCSs). The results indicated that systems managed to protect siRNA from external effects, maintain the system stability, facilitate cellular entry and enhance endosomal escape. It was noted that the transfection efficiency of the MCSs, which were boosted by the presence of cleavable disulfide bond, was ~15% higher than the commercial product RNAiMax while the efficacy of polyplexes alone were similar to the RNAiMax.Article Citation - WoS: 28Citation - Scopus: 29Development of a High-Flux Thin-Film Composite Nanofiltration Membrane With Sub-Nanometer Selectivity Using a Ph and Temperature-Responsive Pentablock Co-Polymer(American Chemical Society, 2019) Bar, Canbike; Çağlar, Nagahan; Uz, Metin; Mallapragada, Surya K.; Alsoy Altınkaya, SacideProducing block co-polymer-based nanofiltration (NF) membranes with sharp molecular weight cutoffs via an efficient method exhibiting persistent size-based separation quality is challenging. In this study, this challenge was addressed by reporting a facile approach to fabricate pentablock co-polymer (PBC)-based thin-film composite (TFC) NF membranes. The PBC, consisting of temperature-responsive Pluronic F127 (PEO-b-PPO-b-PEO) middle blocks and pH-responsive poly(N,N-(diethylamino)ethyl methacrylate) end blocks, were synthesized by atom-transfer radical polymerization. This polymer was then attached electrostatically to the surface of polysulfone/sulfonated polyether-sulfone support membranes fabricated using a non-solvent-induced phase separation technique. The conformational changes of the PBC chains in response to pH and temperature determined the, pure water flux and neutral solute (PEG 1000) rejection performance of TFC membranes. Permeability of the membranes increased from 13.0 +/- 0.63 to 15.9 +/- 0.06 L/m(2).h bar and from 6.7 +/- 0.00 to 13.9 +/- 0.07 L/m(2).h.bar by changing the solution pH from 4 to 8.5 and temperature from 4 to 25 degrees C, respectively. The pH- and temperature-responsive conformational changes did not affect the PEG 1000 rejection and membrane pore radius, which remained constant at similar to 89% and similar to 0.9 nm, respectively. This important finding was attributed to the high grafting density of co-polymer chains, resulting in spatial limitations among the grafted chains. The pore size of similar to 0.9 nm achieved with the proposed membrane design is the smallest size reported so far for membranes fabricated from block copolymers. TFC membranes demonstrated high stability and maintained their flux and rejection values under both static (storage in an acidic solution for up to 1 month) and dynamic (filtering PEG 1000 solution over 1 week) conditions. Pentablock copolymers enable a NF membrane with a sharp molecular weight cutoff suitable for size-selective separations. The membrane fabrication technique proposed in this study is a scalable and promising alternative that does not involve complex synthetic routes.Article Citation - WoS: 12Citation - Scopus: 13Development of Agcl-Tio2 Xerogels Entrapped Antibacterial Polyacrylonitrile Membranes: the Effect of High Salinity Water on Silver Release, Antibiofouling and Antibacterial Efficacies(Elsevier Ltd., 2020) Uz, Metin; Yaşar Mahlıçlı, Filiz; Şeker, Erol; Alsoy Altınkaya, SacideSilver-containing antibacterial membranes are commonly used to control biofouling during bacteria filtration. Unfortunately, fast and uncontrolled release of silver to water is a challenge since this causes mass accumulation of silver in water resources and insufficient long-term antimicrobial effect. To overcome these disadvantages, we propose to add AgCl-TiO2 xerogels (0–0.8 wt%) in the polyacrylonitrile membranes. The long-term silver retaining of the membranes was evaluated by measuring the silver release under filtration of deionized water in the absence and the presence of 1 M NaCl up to 5 days. The antibiofouling and the antibacterial efficacies were determined by measuring the changes in antibacterial activity and DI water flux of the membranes at the end of 5 days of E. coli filtration. The 0.2 wt% AgCl-TiO2 xerogel incorporated polyacrylonitrile membrane demonstrated a constant ~1 ?g of silver release/cm2 per filtration cycle after a total filtration of 0.05 L/cm2 with 1 M NaCl solution. Additionally, it showed antibacterial efficacy and ~100% recovery of deionized water flux by simple backwashing with water after having been used in many E. coli filtration cycles. Thus, this membrane could potentially be used up to ~5.8 years for 8000 h a year for the filtration of high salinity water. Statement of novelty: Silver-containing antibacterial membranes are commonly used to control biofouling during bacteria filtration. Uncontrolled release of silver from the membrane causes massive silver accumulation in water which in turn leads to contamination of water resources and threat to aquatic organisms. Although silver release is strongly influenced by the salinity of water, the release data was collected through filtration of pure DI water or tap water in literature. To overcome the shortcomings of the published studies, we propose to use AgCl-TiO2 xerogels in membranes due to low solubility of AgCl in water and measure the release by filtering high-salinity water. © 2020 Elsevier B.V.Article Citation - WoS: 13Citation - Scopus: 18Stimuli Responsive Polymer-Based Strategies for Polynucleotide Delivery(Cambridge University Press, 2017) Uz, Metin; Alsoy Altınkaya, Sacide; Mallapragada, Surya K.In recent years, stimuli responsive polymer based gene delivery vehicle design for cancer treatment and treatment of other genetic disorders has received extensive attention. Early studies focusing on DNA delivery have been facilitated by functional polymers and this area has seen further growth spurred by recent gene silencing strategies developed for small RNA [i.e., small interfering RNA (siRNA) or micro RNA (miRNA)] delivery. DNA and small RNAs possess analogous properties; however, their explicit differences define the specific challenges associated with the delivery route and the design of functional materials to overcome distinct challenges. Apart from classical gene delivery, the recent advances in genome editing have revealed the necessity of new delivery devices for genome editing tools. A system involving CRISPR (clustered, regularly interspaced, short palindromic repeats) and an endonuclease CRISPR-associated protein 9 (Cas9) coupled with a short, single-guide RNA (sgRNA) has emerged as a promising tool for genome editing along with functional delivery systems. For all these nucleic acid based treatments, the internal or external physiochemical changes in the biological tissue/cells play a major role in the design of stimuli responsive delivery materials for both in vitro and in vivo applications. This review emphasizes the recent advances in the use of pH, temperature, and redox potential-responsive polymers overcoming hurdles for delivery of gene and gene editing tools for both in vitro and in vivo applications. Specifically the chapter focuses on recently proposed delivery strategies, types of delivery systems, and polymer synthesis/modification methods. The recent advances in CRISPR/Cas9-sgRNA technology and delivery are also described in a separate section. The review ends with current clinical trials, concluding remarks, and future perspectives.Article Citation - WoS: 40Citation - Scopus: 47Gelatin-Based 3d Conduits for Transdifferentiation of Mesenchymal Stem Cells Into Schwann Cell-Like Phenotypes(Elsevier Ltd., 2017) Uz, Metin; Büyüköz, Melda; Sharma, Anup D.; Sakaguchi, Donald S.; Alsoy Altınkaya, Sacide; Mallapragada, Surya K.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 × 106 Pa and pore size of ∼150 μm 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 × 106 Pa and pore size of ∼100 μm 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 (∼2.5 pg/mL NGF and ∼0.7 pg/mL GDNF per cell) of neurotrophic factors, while MSCs in macroporous conduits released slightly lower (∼1.5 pg/mL NGF and 0.7 pg/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.Article Citation - WoS: 65Citation - Scopus: 72Effect of Peg Grafting Density and Hydrodynamic Volume on Gold Nanoparticle-Cell Interactions: an Investigation on Cell Cycle, Apoptosis, and Dna Damage(American Chemical Society, 2016) Uz, Metin; Bulmuş, Volga; Alsoy Altınkaya, SacideIn 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/nm2), 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.Article Citation - WoS: 44Citation - Scopus: 60Development of a Novel Strategy for Controlled Release of Lysozyme From Whey Protein Isolate Based Active Food Packaging Films(Elsevier Ltd., 2016) Pekşen Özer, Bahar Başak; Uz, Metin; Oymacı, Pelin; Alsoy Altınkaya, SacideThe purpose of this study is to develop a novel controlled release system based on pH-responsive polyacrylic acid (PAA)/lysozyme (LYS) complexes incorporated within a hydrophilic whey protein isolate (WPI) film matrix for active food packaging applications. Complex formation is simple under benign conditions that are suitable for preserving antimicrobial activity of the lysozyme. In addition, the pH-dependent charge density of complexes allowed a uniform distribution in the matrix. The properties of the complexes such as size, surface charge and hydrophilicity were varied by changing PAA/LYS ratio (0.1 and 0.3 w/w) and PAA molecular weight (2 kDa and 450 kDa). The effects of complex properties as well as mode of lysozyme incorporation into the films (100%-free, 50%-free+50%-PAA/LYS complex and 100%-PAA/LYS complex) on the LYS release rate, activity and antimicrobial efficacy of the films were investigated. The results have shown that ∼100% LYS loading into the complexes is possible regardless of PAA molecular weight or PAA/LYS ratio. Incorporating lysozyme into the film in complexed form extended its release time from less than 24 h up to 500 h and reduced its diffusivity from ∼10−9 to ∼10−13 cm2/s. The films including 50%-free-LYS+50%-PAA/LYS complex showed a 5.7 log reduction in bacterial population within 72 h whereas 100%-free-LYS containing film could not suppress Listeria innocua growth after 24 h. Overall, the results suggest that complexation of lysozyme with weak polyelectrolytes can be used as an effective strategy to achieve a long-lasting antimicrobial effect and that films prepared with such complexes have great potential as food packaging materials.Article Citation - WoS: 17Citation - Scopus: 22Responsive pentablock copolymers for siRNA delivery(Royal Society of Chemistry, 2015) Uz, Metin; Mallapragada, Surya K.; Alsoy Altınkaya, SacideIn 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.Conference Object Citation - Scopus: 3Characterization of Polysulfone Based Hemodialysis Membranes by Afm(Elsevier Ltd., 2012) Uz, Metin; Yaşar Mahlıçlı, Filiz; Polat, Mehmet; Alsoy Altınkaya, SacideMost of the hemodialysis membrane materials are hydrophobic in nature and allow protein adsorption on the surface easily due to hydrophobic interaction between membrane surface and protein molecules when in contact with blood. Adsorbed proteins can affect platelet and leukocyte adhesion, and modulate the response of plasmatic reactions followed by the activation of different defense systems in blood (Sun et al. 2003).