Bioengineering / Biyomühendislik
Permanent URI for this collectionhttps://hdl.handle.net/11147/4529
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Article Citation - Scopus: 3Development of Chrono-Spectral Gold Nanoparticle Growth Based Plasmonic Biosensor Platform(Elsevier, 2024) Sözmen, Alper Baran; Elveren, Beste; Erdoğan, Duygu; Mezgil, Bahadır; Baştanlar, Yalın; Yıldız, Ümit Hakan; Arslan Yıldız, AhuPlasmonic sensor platforms are designed for rapid, label-free, and real-time detection and they excel as the next generation biosensors. However, current methods such as Surface Plasmon Resonance require expertise and well-equipped laboratory facilities. Simpler methods such as Localized Surface Plasmon Resonance (LSPR) overcome those limitations, though they lack sensitivity. Hence, sensitivity enhancement plays a crucial role in the future of plasmonic sensor platforms. Herein, a refractive index (RI) sensitivity enhancement methodology is reported utilizing growth of gold nanoparticles (GNPs) on solid support and it is backed up with artificial neural network (ANN) analysis. Sensor platform fabrication was initiated with GNP immobilization onto solid support; immobilized GNPs were then used as seeds for chrono-spectral growth, which was carried out using NH2OH at varied incubation times. The response to RI change of the platform was investigated with varied concentrations of sucrose and ethanol. The detection of bacteria E.coli BL21 was carried out for validation as a model microorganism and results showed that detection was possible at 102 CFU/ml. The data acquired by spectrophotometric measurements were analyzed by ANN and bacteria classification with percentage error rates near 0% was achieved. The proposed LSPR-based, label-free sensor application proved that the developed methodology promises utile sensitivity enhancement potential for similar sensor platforms. © 2024 The Author(s)Article Citation - WoS: 18Citation - Scopus: 17Modifying Pickering Polymerized High Internal Phase Emulsion Morphology by Adjusting Particle Hydrophilicity(Elsevier, 2024) Durgut, Enes; Zhou, Muchu; Dikici, Betuel Aldemir; Foudazi, Reza; Claeyssens, FrederikThis study investigates the use of submicron polymeric particles with varying crosslinking densities as the sole stabilizer for producing Polymerized High Internal Phase Emulsions (PolyHIPE). We establish a direct correlation between the crosslinking density and the hydrophilicity of the polymer particles. The hydrophilicity of these particles significantly influences the morphology and rheology of HIPEs. These differences manifest as various morphological variations in the resulting PolyHIPE templates. It was discovered that by increasing the crosslinker weight percentage in the particles from 0 % to 100 %, PolyHIPEs with semi-open, open, and closed porous structures can be obtained. Furthermore, non-crosslinked particles were observed to dissolve in the continuous phase, acting as macromolecular surfactants that generate small pores akin to surfactant-stabilized structures in PolyHIPE. These findings offer fresh insights into the relationship between particle localization at the interface, HIPE rheology, and the formation of pore throats in Pickering PolyHIPEs, leading to the creation of either closed or open porous networks. Additionally, interfacial rheological results demonstrate that particles synthesized with varying monomer-to-crosslinker ratios exhibit different interfacial elasticities, which are linked to PolyHIPE morphology.Article Citation - Scopus: 11Μdacs Platform: a Hybrid Microfluidic Platform Using Magnetic Levitation Technique and Integrating Magnetic, Gravitational, and Drag Forces for Density-Based Rare Cancer Cell Sorting(Elsevier, 2023) Keçili, Seren; Yılmaz, Esra; Özçelik, Özge Solmaz; Anıl İnevi, Müge; Günyüz, Zehra Elif; Yalçın Özuysal, Özden; Özçivici, Engin; Tekin, Hüseyin CumhurCirculating tumor cells (CTCs) are crucial indicators of cancer metastasis. However, their rarity in the bloodstream and the heterogeneity of their surface biomarkers present challenges for their isolation. Here, we developed a hybrid microfluidic platform (microfluidic-based density-associated cell sorting (µDACS) platform) that utilizes density as a biophysical marker to sort cancer cells from the population of white blood cells (WBCs). The platform utilizes the magnetic levitation technique on a microfluidic chip to sort cells based on their specific density ranges, operating under a continuous flow condition. By harnessing magnetic, gravitational, and drag forces, the platform efficiently separates cells. This approach involves a microfluidic chip equipped with a microseparator, which directs cells into top and bottom outlets depending on their levitation heights, which are inversely proportional to their densities. Hence, low-density cancer cells are collected from the top outlet, while high-density WBCs are collected from the bottom outlet. We optimized the sorting efficiency by varying the flow rates, and concentrations of the sorting medium's paramagnetic properties using standard densities of polymeric microspheres. To demonstrate the platform's applicability, we performed hybrid microfluidic sorting on MDA-MB-231 human breast cancer cells and U-937 human monocytes. The results showed efficient sorting of rare cancer cells (≥100 cells/mL) from serum samples, achieving a sorting efficiency of ∼70% at a fast-processing speed of 1 mL h−1. This label-free approach holds promise for rapid and cost-effective CTC sorting, facilitating in-vitro diagnosis and prognosis of cancer. © 2023 The Author(s)Article Citation - WoS: 14Citation - Scopus: 16Synergistic Effect of Type and Concentration of Surfactant and Diluting Solvent on the Morphology of Emulsion Templated Matrices Developed as Tissue Engineering Scaffolds(Elsevier, 2022) Claeyssens, Frederik; Aldemir Dikici, Betül; Dikici, SerkanEmulsion templating is an advantageous route for the fabrication of tissue engineering scaffolds. Emulsions are mostly stabilised using surfactants, and the performances of the surfactants depend on various parameters such as emulsification temperature and the presence of the electrolytes. In this study, we suggest that diluting solvent type also has a dramatic impact on the efficiency of the surfactant and morphology of the polymerised emulsions. For this, morphologies of polycaprolactone methacrylate-based polymerised emulsions, which are designed for tissue engineering applications and in vitro biocompatibilities, were shown by our group, prepared using four different surfactants, and three different solvents were investigated. Results showed that the diluting solvent used in the emulsion composition has a strong impact on the performance of the surfactant and consequently on the morphology of polymerised emulsions. Increasing surfactant concentration and diluting solvent volume have an opposite impact on the characteristics of emulsions. Scaffolds with average pore sizes changing from 10 to 78 μm could be fabricated. Establishing these relations enables us to have control over the overall morphology of polymerised emulsions and precisely engineer them for specific tissue engineering applications by tuning solvent and surfactant type and concentration.Article Citation - WoS: 4Citation - Scopus: 6Rational Design of Thermophilic Cyp119 for Progesterone Hydroxylation by in Silico Mutagenesis and Docking Screening(Elsevier, 2023) Kestevur Doğru, Ekin; Güralp, Gülce; Uyar, Arzu; Sürmeli, Nur BaşakSteroid-based chemicals can affect the metabolism, immune functions, and development of sexual characteristics. Because of these effects, steroid derivatives are widely used in the pharmaceutical industry. Progesterone is a steroid-based hormone that mainly controls the ovulation period of women but is also a precursor molecule for the synthesis of important hormones like testosterone and cortisone. Cytochrome P450 (CYP) enzymes are important for the production of hydroxyprogesterones in the industry since they can catalyze regio- and enantioselective hydroxylation reactions. Although human CYP enzymes can catalyze hydroxyprogesterone synthesis with high selectivity, these enzymes are membrane bound, which limits their application for industrial production. CYP119 is a soluble and thermophilic enzyme from the archaea Sulfolobus acidocaldarius. Even though the native substrate of the enzyme is not known, CYP119 can catalyze styrene epoxidation, lauric acid hydroxylation, and Amplex®Red peroxidation. In this work, an in silico mutagenesis approach was used to design CYP119 mutants with high progesterone affinity. Energy scores of progesterone docking simulations were used for the design and elimination of single, double, and triple mutants of CYP119. Among designed 674 mutants, five of them match the criteria for progesterone hydroxylation. The most common mutation of these five mutants, L69G mutant was analyzed using independent molecular dynamics (MD) simulations in comparison with the wild-type (WT) enzyme. L69G CYP119, was expressed and isolated from Escherichia coli; it showed 800-fold higher affinity for progesterone compared to WT CYP119. L69G CYP119 also catalyzed progesterone hydroxylation. The novel designed enzyme L69G CYP119 is a potential versatile biocatalyst for progesterone hydroxylation that is expected to be stable under industrial production conditions.Article Citation - WoS: 17Citation - Scopus: 22Protein Corona Formation on Silver Nanoparticles Under Different Conditions(Elsevier, 2022) Tomak, Aysel; Yılancıoğlu, Buket; Winkler, David; Öksel Karakuş, CeydaThe surfaces of nanoparticles become covered by biomolecules in biological fluids. This protein ‘corona’ modifies materials’ characteristics and biological activity. The composition of the protein corona is dynamic, abundant biomolecules that bind first are subsequently replaced by less abundant but more tightly bound ones. Here, we explore the formation of the silver nanoparticle protein corona on exposure to cell culture media containing 10 % fetal bovine serum supplemented Dulbecco's Modified Eagle's medium. Sodium dodecyl-sulfate polyacrylamide gel electrophoresis and liquid chromatography-mass spectrometry/mass spectrometry analysis were used to monitor how different parameters such as incubation time, heating duration, cell culture medium, incubation temperature, and the number of washes affect the nanoparticle–protein corona complex. silver nanoparticles with and without bound proteins were characterized by electron microscopy, dynamic light scattering, and ultraviolet-visible-near-IR spectroscopy. The tetrazolium-based MTT assay was used to determine viability of A549 human lung adenocarcinoma cells treated with silver nanoparticles. Characterization of the nanoparticles before and after protein binding provided insights into their changing morphology on corona formation. Our results confirmed that the physiological environment directly affects protein corona formation on nanoparticle surfaces. In particular, incubation condition-dependent differences in the amount of bound proteins were observed. This work highlights the importance of environmental drivers of protein adsorption, which should be considered when predicting and/or controlling protein targets of silver nanoparticles.Article Citation - WoS: 11Citation - Scopus: 11The Importance of Neopterin in Covid-19: The Prognostic Value and Relation With the Disease Severity(Elsevier, 2022) Rasmi, Yousef; Heidari, Nadia; Kübra Kırboğa, Kevser; Hatamkhani, Shima; Tekin, Burcu; Alipour, Shahryar; Naderi, Roya; Farnamian, Yeghaneh; Akça, İlknurCoronavirus Disease 2019 [COVID-19], caused by severe acute respiratory syndrome coronavirus 2 [SARS-CoV-2], has rapidly evolved into a global health emergency. Neopterin [NPT], produced by macrophages when stimulated with interferon [IFN-]gamma, is an essential cytokine in the antiviral immune response. NPT has been used as a marker for the early assessment of disease severity in different diseases. The leading cause of NPT production is the pro-inflammatory cytokine IFN-. Macrophage activation has also been revealed to be linked with disease severity in SARS-CoV-2 patients. We demonstrate the importance of NPT in the pathogenesis of SARS-CoV-2 and suggest that targeting NPT in SARS-CoV-2 infection may be critical in the early prediction of disease progression and provision of timely management of infected individuals.Article Citation - Scopus: 6Sensitive and Rapid Protein Assay Via Magnetic Levitation(Elsevier, 2022) Sözmen, Alper Baran; Arslan Yıldız, AhuMagnetic levitation (MagLev) is a newly emerging methodology for biosensing that provides a density-based analysis, which is highly sensitive and versatile. In this study, a magnetic levitation based sensor platform was used for protein detection; and sensor platform optimization was performed for both sensitivity and resolution. Bovine Serum Albumin (BSA) was used as a model protein and detection of BSA was carried out by antibody functionalized polystyrene microspheres (PSMs). Various sizes of PSMs were examined and their performances were compared by statistical analyses in terms of limit of detection (LOD), sensitivity, and resolution. Quantification of the protein was done based on the magnetic levitation height differences of antibody functionalized PSMs. For optimization of the methodology, varied PSMs were utilized, and standardization of PSM diameter, concentration of the antibody to be functionalized, and PSM dilution rates were carried out. In conclusion, 20 μm PSMs diluted to 0.005% W/V and functionalized with anti-BSA antibody at a concentration of 28 μg/ml were determined to provide the best resolution for BSA detection. A dynamic range of 100 nM to 1 mM was observed with an LOD value of 4.1 ng/ml. This sensing platform promises a novel approach with a diverse application field and it provides rapid, consistent, and reproducible results with high resolution and sensitivity.Article Citation - WoS: 8Citation - Scopus: 9Optimization of the Algal Species Chlorella Miniata Growth: Mathematical Modelling and Evaluation of Temperature and Light Intensity Effects(Elsevier, 2022) Sözmen, Alper Baran; Ata, Ayça; Övez, BikemGrowth of Chlorella miniata, a green microalga was investigated during this study under various temperature and light intensity values with the purpose of determining growth rate changes of the microalgae with cultivation parameters, experiments were carried out using airlift photobioreactors with a study volume of 6 L. Culturing conditions were between 66 and 385 μmol photon m−2 s−1 and between 14 and 30 °C for light intensity and ambient temperature, respectively. Acquired data were then used to test various mathematical models for coherency with experimental results. Aiba Model for light intensity and Skewed Normal Distribution Model for temperature parameters performed superior compared to the rest of the mathematical models used during the study. Utilizing both mathematical models a novel model was deduced to express effects of both light intensity and temperature parameters in combination on algal growth. Then the developed model was used to calculate the optimum growth condition of the species. The proposed mathematical model showed good coherency with experimental data and an average relative error of 1.97% for both temperature and light intensity effects on algal growth. The theoretical optimum temperature and light intensity for the maximum specific growth rate were calculated to be 22.43 °C and 291.5 μmol photon m−2 s−1 respectively.Article Citation - WoS: 11Citation - Scopus: 11Absorbance-Based Detection of Arsenic in a Microfluidic System With Push-And Pumping(Elsevier, 2021) Karakuzu, Betül; Gülmez, Yekta; Tekin, H. CumhurRapid and portable analysis of arsenic (As) contamination in drinking water is very important due to its adverse health effects on humans. Available commercial detection kits have shown low sensitivity and selectivity in analysis, and also they can generate harmful by-products. Microfluidic-based approaches allow portable analysis with gold nanoparticles (AuNPs) as labels. However, they need complex surface modification steps that complicate detection protocols. Due to the lack of precise sensing and affordable solution, we focused on developing a microfluidic platform that uses a push-and-pull pumping method for sensitive detection of As. In this detection principle, a sample is introduced in the microfluidic channel modified with -SH functional groups where As can bind. Then, AuNPs are given in the channel and AuNPs bind on free -SH functional groups which are not allocated with As. Absorbance measurements are conducted to detect AuNPs absorbed on the surfaces and the resulting absorbance value is inversely proportional with As concentration. The method enables detection of As down to 2.2 mu g/L concentration levels in drinking water, which is well-below the allowed maximum As concentration of 10 mu g/L in the drinking waters by the World Health Organization (WHO). The paper reveals that multiple push-and-pull pumping of fixed volume of sample and AuNPs with a syringe pump can improve the binding efficiency in the microfluidic channel. With this technique, low amounts of sample (1 mL) and short total assay time (25 min) are sufficient to detect As.