Master Degree / Yüksek Lisans Tezleri
Permanent URI for this collectionhttps://hdl.handle.net/11147/3008
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Master Thesis Development and Characterization of Magnesium Alginate Hydrogels for 3d Cell Culture Formation(01. Izmir Institute of Technology, 2021) Çoban, Başak; Arslan Yıldız, AhuCell culture is an important tool for biological research. Two-dimensional (2D) cell culture is still used but growing cells on plastic surfaces offering unnatural growth kinetics and cell attachment. Three-dimensional (3D) cell culture allows cells to growth in their 3D physical shape and interact with their surroundings which represent the natural microenvironment. Hydrogels are crosslinked networks, have become increasingly used biomaterial for 3D cell culture with their ability to simulate the nature of most soft tissues. In this thesis, a new methodology based on bio-patterning was developed to fabricate (3D) cellular structures by using Mg-alginate hydrogel and fabricated 3D cellular structures was utilized for drug screening studies. Mg-alginate hydrogel has a specific gelation/de-gelation characteristics compared to other types of hydrogels due to its weak polymer-ion interaction. In this study slow gelation and de-gelation property of Mg-alginate hydrogel was used for biopatterning of 3D cellular structures. Plackett-Burman and Box-Behnken design models were used to optimize parameters of Mg alginate-based biopatterning method while using HeLa cells as a model cell line. Then, the applicability of newly developed methodology was successfully demonstrated by using SaOS-2 and SH-SY5Y cells to fabricate 3D cellular structures. Cell proliferation and migration profiles were observed during long-term culturing with time-dependent light microscopy images. Also cell proliferation and viability of long-term cultured tumor models were analyzed by using Alamar Blue and Live/Dead assays. Moreover, F-actin, Collagen I, and DAPI staining/immunostaining was done to investigate cellular and extracellular components of 3D cellular structures for short and long-term culture times. Finally, the dose-response of fabricated 3D structures was evaluated and compared with standard 2D cell culture by applying doxorubicin (DOX). The IC50 values were calculated for 3D cellular structure of HeLa, SaOS-2 and SH-SY5Y cells as 8.2, 7.8, and 2.1 µM respectively while IC50 values of 2D controls obtained as 3.2, 4.4, and 0.2 µM respectively. These results were also statistically analyzed and dose responses were found significantly different according to t-test, which means 3D cellular structures were more resistant to drug exposure compared to 2D cell culture.Master Thesis Characterization and Utilization of Injectable Hydrogels for Tissue Engineering Applications(Izmir Institute of Technology, 2020) Güzelgülgen, Meltem; Arslan Yıldız, AhuTissue engineering combines the knowledge of the engineering aspects with life sciences to improve human health. Recent studies in tissue engineering have focused on investigating biocompatible scaffold materials and design. Quince seed hydrogel(QSH) has been used in traditional and modern medicine for skin wound and burn treatments, synovial lubrication, cough and asthma removal, and oral drug delivery with its antioxidant potential and biocompatible aspects. This thesis focuses on developing QSH and evaluating its potential as an injectable hydrogel in treating bone tissue defects as a totally new tissue scaffold and also as a promising tissue filling material. For this purpose, QSH scaffold optimization was carried out using various concentrations of hydrogel and crosslinkers which were glutaraldehyde(GTA) and 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide(EDC)/N-hydroxysuccinimide(NHS). Morphological and chemical analysis of QSH was done using SEM, FTIR, AFM, and protein adsorption test. Thus, porosity, swelling ratio, degradation rate and surface characteristics were evaluated. NIH-3T3 and SaOS-2 cell lines were utilized for 3D cell culture formation. Afterward, 3D spheroids were analyzed for cell viability and proliferation by using AlamarBlue and LiveDead assays, and also cell imaging technics. Results showed that QSH scaffolds did not show any cytotoxic effect on NIH-3T3 and SaOS-2 cells. The optimum results were achieved with 2mg/mL of QSH and 0.03M GTA concentrations; where 76.59µm average pore size, 56.8 fold water holding capacity and at least 80% cell viability was observed. Therefore, it was concluded that QSH has a high potential to promote tissue engineering applications with its injectable texture as a filling material.Master Thesis Hydrogels and Self-Assemled Nanostructures Based on Wool Keratose(Izmir Institute of Technology, 2017) Pakkaner, Efecan; Top, AybenIn this study, water soluble keratose proteins were extracted from “Ovis aries” wool using peracetic acid oxidation with a yield of 35 ± 5 %. Wool samples and the extracted keratose proteins were characterized by using FT-IR, XRD, SEM and TGA techniques. α-keratose fractions (MW = 43-53 kDa) along with cleaved fragments of α-keratoses with molecular weights between 23 and 33 kDa were identified in the extracted protein mixture using SDS-PAGE analysis. DLS and AFM experiments indicated self-assembled globular nanoparticles with diameters of 20-40 nm formed at 5 and 10 mg/ml keratose concentrations. On the other hand, at 10 % w/v keratose concentration interconnected keratose hydrogels with pore sizes of 6 ± 4 and 7 ± 4 μm were obtained upon incubation at 37 and 50 °C, respectively. Storage moduli (G’) of these physical hydrogels were increased from ~100 to ~1000 Pa, as gelation temperature was increased from 37 to 50 °C. Hydrogels were also obtained at 7.5 % w/v keratose concentration by the addition of a crosslinker, THPC. Amine group:crosslinker ratio was used as 1:1, 1:2 and 1:4. As the amount of crosslinker increased, network transformed from fibrous to more planar structures exhibiting a significant decrease in average pore size from 24 to 11 μm. G’ values of the crosslinked hydrogels were obtained between ~1 and ~5 kPa tuned by the crosslinking amount. Cell interaction properties of a select physical hydrogel prepared at 37 °C was tested using CCK-8 assay. It was observed that the keratose hydrogel supported L929 mouse fibroblast cell proliferation as much as collagen, which suggests that these keratose hydrogels can be promising candidates in soft tissue engineering applications.