Bioengineering / Biyomühendislik
Permanent URI for this collectionhttps://hdl.handle.net/11147/4529
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Conference Object A Glucuronoxylan-Based Bio-Ink Development: Characterization and Application(Wiley, 2023) Yıldırım, Ömer; Arslan Yıldız, AhuBioprinting is a trending technique that enables the fabrication of threedimensional (3D) constructs in designed shapes and with desired properties. Bioinks are one of the most significant components of bioprinting and the successful fabrication of 3D bioprinted constructs mostly depends on the features of bioinks that would be used. New generation bioinks that are soft and viscous enough, printable under low pressure, stable in cell culture, and have fast gelation mechanisms are ideal to be used in current bioprinting techniques. Hydrocolloids have said features and have similar properties to native ECM structures. Hence bioinks that are developed from hydrocolloids can be utilized for mimicking of ECM structure of soft tissues. Polysaccharidebased hydrocolloids are ideal bioink candidates with their high waterholding capacity and biocompatibility. Here, a glucuronoxylanbased newgeneration bioink was developed, and its printability was evaluated for 3D bioprinting applications. The glucuronoxylanbased hydrocolloid was obtained by water extraction of quince seeds and its utilization in bioprinting was investigated. Bioink characterization was done by FTIR and mechanical analysis. Bioprinting parameters were optimized assessing uniformity, pore factor, and shape fidelity. Then, the characterization of bioprinted constructs was performed by pore angle measurement, waterholding capacity analysis, protein adsorption, and cell viability assays. Bioprinted structures have high mechanical strength, suitable protein adsorption behavior, and waterholding capacity as high as 20fold of its own weight, which is higher than other hydrogels that were used in soft tissue engineering. Moreover, the cell viability results of fibroblast cells in the bioink were high for longterm culture. In conclusion, findings show that the developed glucuronoxylanbased bioink is a biocompatible and promising bioink material for further tissue engineering applications.Article Citation - WoS: 4Citation - Scopus: 5Antioxidant and Antimicrobial Activities of Plants Grown in the Mediterranean Region(John Wiley and Sons Inc., 2022) Kaçar, D.; Bayraktar, Oğuz; Erdem, C.; Alamri, A.S.; Galanakis, C.M.Background: The main objective of this research was to identify plant species with possible bioactivities based on their total phenol content, antioxidant, and antimicrobial properties. Therefore, different parts of 42 plant species grown in the Mediterranean region were extracted with aqueous ethanol solutions to prepare extracts with antioxidant and antimicrobial activities, mainly resulting from their total phenol contents. No detailed laboratory data on the flora of this area exists regarding their total phenol contents and total antioxidant activities. Results: Yields of extraction for each plant material were determined. Extracts were characterized based on their total phenol contents, total antioxidant (both hydrophilic and lipophilic), and antimicrobial activities using Folin–Ciocalteu, Photochemiluminescence, disc diffusion, and microdilution methods, respectively. The extract of Hypericum empetrifolium had the relatively highest total water-soluble and lipid-soluble antioxidant activities. Sarcopoterium spinosum extract had relatively high total phenol content. Preliminary screening study was conducted with the disc diffusion method to evaluate the extracts' antimicrobial activities. 26 out of 42 plant species showed significant antimicrobial activities against the growth of microorganisms. Microdilution assays were performed to evaluate the most active plant species with their minimum inhibition concentrations. H. empetrifolium, Pistacia terebinthus, Arbutus unedo, and Cistus parviflorus were the most antimicrobial plant species among those investigated. CONCLUSION: The new potential sources for the isolation of bioactive natural compounds from specific plant species could be possible with the help of this present screening study. Isolated bioactive natural compounds can be utilized as raw materials in cosmetics, nutraceuticals, food supplements, and pharmaceutical industries. © 2022 Society of Chemical Industry.Conference Object Biopatterning of 3d Cellular Structures Via Contactless Magnetic Manipulation for Drug Screening(Mary Ann Liebert, 2023) Onbas, Rabia; Arslan Yıldız, AhuArticle Citation - WoS: 3Citation - Scopus: 4Biopatterning of 3d Cellular Model by Contactless Magnetic Manipulation for Cardiotoxicity Screening(Mary Ann Liebert, Inc, 2023) Önbaş, Rabia; Arslan Yıldız, AhuPatterning cells to create three-dimensional (3D) cell culture models by magnetic manipulation is a promising technique, which is rapid, simple, and cost-effective. This study introduces a new biopatterning approach based on magnetic manipulation of cells with a bioink that consists alginate, cells, and magnetic nanoparticles. Plackett-Burman and Box-Behnken experimental design models were used to optimize bioink formulation where NIH-3T3 cells were utilized as a model cell line. The patterning capability was confirmed by light microscopy through 7 days culture time. Then, biopatterned 3D cardiac structures were formed using H9c2 cardiomyocyte cells. Cellular and extracellular components, F-actin and collagen Type I, and cardiac-specific biomarkers, Troponin T and MYH6, of biopatterned 3D cardiac structures were observed successfully. Moreover, Doxorubicin (DOX)-induced cardiotoxicity was investigated for developed 3D model, and IC50 value was calculated as 8.1 μM for biopatterned 3D cardiac structures, which showed higher resistance against DOX-exposure compared to conventional two-dimensional cell culture. Hereby, developed biopatterning methodology proved to be a simple and rapid approach to fabricate 3D cardiac models, especially for drug screening applications. Copyright 2023, Mary Ann Liebert, Inc., publishers.Conference Object Biofabrication of Scaffold-Free 3d Cellular Structures Using Magnetic Levitational Assembly To Study Cardiac Toxicity(Mary Ann Liebert, 2023) Yıldız, Ahu Arslan; Arslan Yıldız, Ahu; Onbaş, RabiaSpheroids are one of the well-characterized 3D cell culture approaches for drug screening and therapeutic studies. Magnetic levitation (MagLev) is a newly developing approach to form 3D cellular structures and spheroids [1,2,3]. Magnetic levitational assembly of cells provides rapid, simple, cost-effective 3D cell culture formation while ensuring scaffold-free microenvironment. Here, our efforts are summarized in designing new magnetic levitation platform and biofabrication of 3D cellular entities via magnetic levitation for tissue engineering. Magnetic levitation and guidance of cells were provided by using a paramagnetic agent to fabricate scaffold-free 3D cellular structures. The parameters of cell density, paramagnetic agent concentration, and culturing time were optimized to obtain 3D cardiac cellular structures with tunable size, circularity, and high cell viability. Cellular and extracellular components of the 3D cellular structures were demonstrated via immunofluorescent staining. Also, 3D cardiac cellular structures showed more resistance to drug exposure compared to 2D control. In conclusion, MagLev methodology offers an easy and efficient way to fabricate 3D cellular structures for drug screening studies.Conference Object Biopatterning of 3d Cellular Structures Via Contactless Magnetic Manipulation for Drug Screening(Mary Ann Liebert, 2023) Önbaş, Rabia; Arslan Yıldız, Ahu"Patterning and manipulation techniques have been used to fabricate 3D cell cultures in tissue engineering. The contactless magnetic manipulation approach is a rapid, simple, and cost-effective method that requires paramagnetic agents [1-3] or magnetic materials [4]. Here, to obtain patterned 3D cellular structures a new alginate-based bio-ink formulation was developed to fabricate 3D cellular structures using contactless magnetic manipulation. 3D cardiac model was obtained by patterning rat cardiomyocytes. Cellular and extracellular components and cardiac-specific markers of patterned 3D cellular structures were indicated successfully. Drug response of patterned 3D cellular structures was evaluated by applying doxorubicin. Patterned 3D cardiac cellular structures showed significantly different drug response compared to conventional 2D cell cultures. In conclusion, this technique provides an easy, efficient, and low-cost methodology to fabricate 3D cardiac structures for drug screening.Article Citation - WoS: 6Citation - Scopus: 6Functional Characterization of a Novel Cyp119 Variant To Explore Its Biocatalytic Potential(Wiley, 2021) Sakallı, Tuğçe; Sürmeli, Nur BaşakBiocatalysts are increasingly applied in the pharmaceutical and chemical industry. Cytochrome P450 enzymes (P450s) are valuable biocatalysts due to their ability to hydroxylate unactivated carbon atoms using molecular oxygen. P450s catalyze reactions using nicotinamide adenine dinucleotide phosphate (NAD(P)H) cofactor and electron transfer proteins. Alternatively, P450s can utilize hydrogen peroxide (H2O2) as an oxidant, but this pathway is inefficient. P450s that show higher efficiency with peroxides are sought after in industrial applications. P450s from thermophilic organisms have more potential applications as they are stable toward high temperature, high and low pH, and organic solvents. CYP119 is an acidothermophilic P450 from Sulfolobus acidocaldarius. In our previous study, a novel T213R/T214I (double mutant [DM]) variant of CYP119 was obtained by screening a mutant library for higher peroxidation activity utilizing H2O2. Here, we characterized the substrate scope; stability toward peroxides; and temperature and organic solvent tolerance of DM CYP119 to identify its potential as an industrial biocatalyst. DM CYP119 displayed higher stability than wild-type (WT) CYP119 toward organic peroxides. It shows higher peroxidation activity for non-natural substrates and higher affinity for progesterone and other bioactive potential substrates compared to WT CYP119. DM CYP119 emerges as a new biocatalyst with a wide range of potential applications in the pharmaceutical and chemical industry.Article Citation - WoS: 2Citation - Scopus: 2Cloning, Expression, and Characterization of a Novel Sericin-Like Protein(Wiley, 2022) Bostan, Fatmanur; Sürmeli, Nur BaşakSilk consists of two proteins called fibroin and sericin. While fibroin is used in the textile industry and has various biomaterial applications, sericin has been considered as waste material until recently. Sericin is a multicomponent protein and it has important properties such as biocompatibility, biodegradability, cryoprotectivity, and antioxidant. Sericin from silkworm cocoons can be obtained by chemical, enzymatic, and heat treatment methods. However, sericin obtained with these treatment methods is not of consistent and high quality. Moreover, the exposure of sericin to harsh conditions during extraction leads to inconsistencies in the composition and structure of the sericin obtained. The inconsistencies in sericin structure and composition decrease application of sericin as a biomaterial. Here, we produce a sericin-like protein (Ser4mer) with native sequence of sericin encoding four repeats of the conserved 38 amino acid motif recombinantly in Escherichia coli and characterize its structural properties. Ser4mer protein shows similar structure to native sericin and higher solubility than previously obtained recombinant sericin-like proteins. Recombinant production of a soluble sericin-like protein will significantly expand its applications as a biomaterial. In addition, recombinant production of silk proteins will allow us to understand sequence-structure relationships in these proteins.Article Citation - WoS: 27Citation - Scopus: 28Biocomposite Scaffolds for 3d Cell Culture: Propolis Enriched Polyvinyl Alcohol Nanofibers Favoring Cell Adhesion(John Wiley and Sons Inc., 2021) Bilginer, Rumeysa; Özkendir İnanç, Dilce; Yıldız, Ümit Hakan; Arslan Yıldız, AhuThe objective of this work is generation of propolis/polyvinyl alcohol (PVA) scaffold by electrospinning for 3D cell culture. Here, PVA used as co-spinning agent since propolis alone cannot be easily processed by electrospinning methodology. Propolis takes charge in maximizing biological aspect of scaffold to facilitate cell attachment and proliferation. Morphological analysis showed size of the electrospun nanofibers varied between 172-523 nm and 345-687 nm in diameter, for non-crosslinked and crosslinked scaffolds, respectively. Incorporation of propolis resulted in desired surface properties of hybrid matrix, where hybrid scaffolds highly favored protein adsorption. To examine cell compatibility, NIH-3T3 and HeLa cells were seeded on propolis/PVA hybrid scaffold. Results confirmed that integration of propolis supported cell adhesion and cell proliferation. Also, results indicated electrospun propolis/PVA hybrid scaffold provide suitable microenvironment for cell culturing. Therefore, developed hybrid scaffold could be considered as potential candidate for 3D cell culture and tissue engineering.Article Citation - WoS: 5Citation - Scopus: 7Five New Cardenolides Transformed From Oleandrin and Nerigoside by Alternaria Eureka 1e1bl1 and Phaeosphaeriasp. 1e4cs-1 and Their Cytotoxic Activities(Elsevier Ltd., 2021) Karakoyun, Çiğdem; Küçüksolak, Melis; Bilgi, Eyüp; Doğan, Gamze; Çömlekçi, Yiğit Ege; Bedir, ErdalBiotransformation of oleandrin (1) and nerigoside (2) by endophytic fungi; Alternaria eureka 1E1BL1 and Phaeospheria sp. 1E4CS-1, has led to the isolation of five new metabolites (3, 5, 6, 7 and 8) together with a known compound (4). The structures of the biotransformation products were elucidated by 1D-, 2D NMR and HR-MS. Phaeospheria sp. mainly provided monooxygenation reactions on the A and B rings, whereas A. eureka afforded both monooxygenated and desacetylated derivatives of the substrates. Cytotoxic activity of the compounds was tested against a non-cancerous (HEK-293) and four cancer (PANC-1, MIA PaCa-2, DU 145 and A549) cell lines by MTT cell viability assay. All compounds were less cytotoxic than oleandrin, which had IC50 values ranging between 2.7 and 41.9 nM. Two of the monohydroxylated metabolites, viz. 7(?)-hydroxy oleandrin (3) and 1(?)-hydroxy oleandrin (7), were also potent with IC50 values from 18.45 to 39.0 nM, while desacetylated + monohydroxylated, or dihydroxylated products had much lower cytotoxicity. Additionally, the lesser activity of 2 and its metabolite (6) possessing diginose as sugar residue inferred that oleandrose moiety is important for the toxicity of oleandrin as well as hydrophobicity of the steroid core. © 2020 Phytochemical Society of Europe