Bioengineering / Biyomühendislik
Permanent URI for this collectionhttps://hdl.handle.net/11147/4529
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Research Project Birden fazla biyomolekülün algılanması için akıllı nanoyapı dizileri(2014) Zareie, Hadi M.; Bulmuş, VolgaBiyolojik maddelerin hızlı, nicel ve paralel bir şekilde algılanabilmesine, biyomedikal, çevre, biyoteknoloji, savunma ve tarım gibi birçok alanda ihtiyaç duyulmaktadır. Birim kütle başına çok geniş yüzey alanına sahip olan nanomalzemeler boyuta ve şekle bağlı eşsiz kimyasal ve fiziksel özellikler sergilerler. Bu özellikleri sayesinde nanomalzemeler, biyolojik numunelerin algılanmasında son derece duyarlı, etikete/işaretlemeye ihtiyaç duymayan, hızlı metotlar geliştirmek için eşsiz fırsatlar sunmaktadır. Bu projede hedefimiz, birden fazla biyolojik maddenin etikete/işaretlemeye ihtiyaç duymadan, nicel ve paralel şekilde algılanması için duyarlı ve hızlı bir sistemin geliştirilmesine yönelik olarak sıcaklık-duyarlı polimerler ve biyomoleküller ile fonksiyonelleştirilmiş sandviç-benzeri nanoyapıların desenli dizilerinin üretilmesi ve çoklu biyomolekül bağlanma olaylarının bir fonksiyonu olarak lokalize yüzey plazmon rezonansının (LSPR) ve kapasitansının incelenmesidir. Bu amaçla öncelikle sandviç-benzeri nanoyapıların desenli dizinleri nanoküre litografisi tekniği ile üretilmiştir. Sandviç-benzeri nanoyapılar, metal-yalıtkan-metal üçlü tabakalardan oluşturulmuştur. Bu nanosandviç dizileri, model biyomoleküller (biyotin, glutatiyon ve tek-sarmal oligoadenin) ve sıcaklıkduyarlı polimer ile fonksiyonelleştirilmiştir. Nanosandviç dizilerinin hazırlanması ve yüzey modifikasyonları, taramalı elektron mikroskobu, atomik kuvvet mikroskopisi, UV-görünür-yakın kızılötesi spektrofotometrisi aracılığıyla LSPR ölçümleri, X-ışını fotoelektron spektrometresi (XPS) gibi farklı teknikler ile doğrulanmıştır. Farklı transdüksiyon mekanizmalarını incelemek için, polimerler ve biyomoleküller ile fonksiyonelleştirilmiş nanodizilerin lokalize yüzey plazmon rezonansı ve kapasitansı, çoklu biyomolekül bağlanma olaylarının bir fonksiyonu olarak ölçülmüştür. LSPR ve XPS kapasitans ölçümleri, nanodizilerin işlevselleştirilmesi adımlarını ve daha da önemlisi sıcaklık kontrollü olarak biyotin-streptavidin ve oligoadenin-oligotimin veya biyotin-streptavidin ve glutatiyon-glutatiyon Stransferaz biyotanıma olaylarını açıkça göstermiştir. Sonuç olarak, bu projenin çıktısı biyomedikal, çevre, biyoteknoloji, tarım, savunma ve benzer endüstrilerde doğrudan uygulamaları olacak yeni ve iyileştirilmiş biyosensörlerin geliştirilmesine yönelik yeni nanomalzemeler ve yöntemler olmuştur.Article Citation - WoS: 2Citation - Scopus: 2Evaluating the Performance of Conventional Daf and Posidaf Processes for Cyanobacteria Separation at a Pilot Plant Scale(IWA Publishing, 2022) Yap, Russell K.L.; Rao, N. R.H.; Holmes, M.; Whittaker, Michael; Stuetz, Richard M.; Jefferson, Bruce; Bulmuş, Volga; Peirson, William Leslie; Henderson, R. K.In this work, a commercially available water treatment polymer poly(N,N-diallyl-N,N-dimethylammonium chloride) (PDADMAC) and a hydrophobically modified polymer (HMP) designed to adhere to bubble surfaces were applied for the first time in the novel Posi-dissolved air flotation process (PosiDAF) that uses polymer-modified bubbles, at pilot-scale for the treatment of waste stabilisation pond samples rich in algae. It was found that PDADMAC in PosiDAF gave comparable removal to that achieved using conventional DAF at .95% cell separation. Furthermore, the float layer was more uniform and thicker with up to 8% solid contents compared to conventional DAF, which comprised discrete floc clusters with an average solid concentration of ∼4.1%. In contrast to the use of PDADMAC, the application of the HMP did not achieve similarly good separation at pilot scale. It was hypothesised that this may be due to the micellisation of the HMP on the bubble surface, creating unstable bubbles that coalesced and prevented polymer-bubble-cell interactions, which are crucial for effective cell separation. On comparison of the costs of PosiDAF and conventional DAF, it was found that PosiDAF resulted in cost-savings of up to 74% due to low chemical consumption. In summary, PosiDAF reduced chemical cost and increased solid contents in the metal-free float.Article Citation - Scopus: 1A Diaminoethane Motif Bearing Low Molecular Weight Polymer as a New Nucleic Acid Delivery Agent(Elsevier, 2021) Zelcak, Aykut; Ünal, Yağmur Ceren; Meşe, Gülistan; Bulmuş, VolgaAmong polymer-based gene delivery systems, poly(ethylene imine) (PEI) stands out as an effective polycation. However, the toxic effects of PEI especially at higher molecular weights limit its usage. Although the effects of PEI's architecture and molecular weight on gene delivery is controversial in literature, low molecular weight PEI appears to be efficient at transfection while having lower toxicity. Herein, as an alternative to low molecular weight, linear PEI, a methacrylate polymer bearing diamimoethane motifs, poly(2-((2-aminoethyl)amino)ethyl methacrylate) (P(AEAEMA)), was evaluated in vitro as a new nucleic acid delivery agent. P(AEAEMA) (8 kDa) showed low toxicity on Skov-3-luc and NIH/3T3 cell lines at polymer concentrations where PEI (8 kDa) was highly toxic. P(AEAEMA) could efficiently form complexes with siRNA at an N/P ratio of 2 as shown by gel electrophoresis. The diameter of P(AEAEMA)-siRNA complexes was found to be significantly lower than PEIsiRNA complexes almost at all tested N/P ratios. P(AEAEMA) could improve the stability of siRNA in serum containing media by protecting the siRNA against serum nucleases. siRNA and pDNA transfection efficiency of P (AEAEMA) on luciferase expressing Skov-3-luc cell line and HEK 293T cell line, respectively was found to be comparable to well-known nucleic acid carrier, PEI. The transfection efficiency of both P(AEAEMA) and PEI was found to be cell-type-dependent. None of the polymers were able to transfect MDA-MB-231 cells with siRNA or pDNA.Article Citation - WoS: 5Citation - Scopus: 6Efficient Synthesis of Crgd Functionalized Polymers as Building Blocks of Targeted Drug Delivery Systems(Elsevier Ltd., 2018) Thankappan, Hajeeth; Zelçak, Aykut; Taykoz, Damla; Bulmuş, VolgaSynthetic peptides with cyclic arginine-glycine-aspartate motif (cRGD) play an important role in cell recognition and cell adhesion. cRGD-decorated soluble polymers and polymeric nanoparticles have been increasingly used for cell-specific delivery of antitumor drugs. While the significance of cRGD modification for tumor cell-specific targeting of polymeric carriers is well-accepted, straightforward procedures ensuring the fidelity of cRGD modification of polymeric systems are still lacking. Herein, we have reported an in-situ polymerization approach for synthesis of cRGD-end-functionalized well-defined polymers as potential building blocks of targeted drug delivery systems. A new cRGD peptide functionalized RAFT agent was synthesized as confirmed by MALDI-TOF and 1H NMR spectroscopy. The ability of this RAFT agent to control polymerizations was then tested using two different monomers oligoethyleneglycol acrylate and t-butyl methacrylate. The RAFT-controlled character of polymerizations and the living characteristic of the synthesized polymers were investigated through a series of kinetic experiments. The cytotoxicity and targeting capability of cRGD-functionalized OEGA polymers were investigated using cell lines expressing αvβ3 integrins at varying extents.Article Citation - WoS: 36Citation - Scopus: 38Effect of Molecular Architecture on Cell Interactions and Stealth Properties of Peg(American Chemical Society, 2017) Özer, İmran; Tomak, Aysel; Zareie, Hadi M.; Baran, Yusuf; Bulmuş, VolgaPEGylation, covalent attachment of PEG to therapeutic biomolecules, in which suboptimal pharmacokinetic profiles limiting their therapeutic utility are of concern, is a widely applied technology. However, this technology has been challenged by reduced bioactivity of biomolecules upon PEGylation and immunogenicity of PEG triggering immune response and abrogating clinical efficacy, which collectively necessitate development of stealth polymer alternatives. Here we demonstrate that comb-shape poly[oligo(ethylene glycol) methyl ether methacrylate] (POEGMA), a stealth polymer alternative, has a more compact structure than PEG and self-organize into nanoparticles in a molecular weight dependent manner. Most notably, we show that comb-shape POEGMA promotes significantly higher cellular uptake and exhibits less steric hindrance imposed on the conjugated biomolecule than PEG. Collectively, comb-shape POEGMA offers a versatile alternative to PEG for stealth polymer-biomolecule conjugation applications.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.