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Öğe Antiadipogenic and antiobesogenic effects of pterostilbene in 3T3-L1 preadipocyte models(TUBITAK, 2023) Gülnar, Birgül; Canatar, İpek; Özdaş, SibelSince obesity causes at least 2.8 million death each year and is a major risk factor for many diseases, it is critical to evaluate alternative treatment approaches. In this context, studies on the research of natural product-based therapeutics in the fight against obesity are increasing. In this study, it was aimed to evaluate the antiadipogenic and antiobesogenic efficacy of pterostilbene a natural phenolic compound in 3T3-L1 cells. The mature 3T3-L1 adipocytes were exposed to pterostilbene at different concentrations and half-maximum inhibitory concentrations (IC50 ) were determined by MTT assay. Oil-Red-O staining was applied to determine lipid accumulation. Phase contrast microscopy, Giemsa, F-actin and DAPI staining were applied to examine the efficacy of pterostilbene on the morphology of 3T3-L1 adipocyte cells. Moreover, expressions of adinopectin and glucose transporter-4 (Glut-4) in relation to insulin resistance were evaluated using immunofluorescent staining and qRT-PCR. Pterostilbene caused no significant cytotoxicity towards preadipocytes at concentrations ?7.5 ?M and the percentage of viable cells remained above about 86% for 24 h, 48 h and 72 h (p > 0.05). Therefore, pterostilbene treatment at 5 and 7.5 ?M was used in the subsequent experiments as safe dosages. In addition, it was observed that pterostilbene treatment reduced lipid accumulation in adipocyte differentiation. Adipocytes treated with a dose of 7.5 µM for 14 days showed less intense lipid deposition and a more spindle-like morphology compared to the adipocytes treated with a dose of 5 µM. Especially on the 14th day, actin filaments were filamentous in adipocytes treated with pterostilbene 7.5 µM compared to the adipocytes treated with a dose of 5 µM; the filaments were similarly oriented as in preadipocytes, and chromatin condensation was observed to be quite high. Our data suggests that the pterostilbene supplementation may help weight control and the antiadipogenic and that antiobesogenic activity is mediated in part by reduction of lipid accumulation and induction of Glut-4 and Adiponectin levels. © TÜBİTAK.Öğe Effects of Knockdown of XPO5 by siRNA on the Biological Behavior of Head and Neck Cancer Cells(John Wiley and Sons Inc, 2022) Özdaş, Sibel; Canatar, İpek; Özdaş, TalihObjectives/Hypothesis: Dysregulated expression of microRNAs (miRNAs) and dysregulation of the mechanisms that regulate them are associated with carcinogenesis. Exportin-5 (XPO5), a member of the Karyopherin family, is responsible for the transfer of pre-miRNAs from the nucleus to the cytoplasm. Despite the high oncogenic potential of XPO5 as a critical regulator of the biogenesis of miRNAs, its role in head and neck squamous cell carcinoma (HNSCC) biology has not been explained yet. Study Design: In-vitro translational. Methods: The expression of XPO5 at the mRNA, protein, and intracellular level in SCC-9, FaDu SCC-90, and Detroit-562 cell lines were evaluated with quantitative reverse transcription polymerase chain reaction, Western-blot analysis, and immunofluorescence staining, respectively. The functional role of XPO5 in HNSCC was analyzed by silencing the gene expression with XPO5-small interfering RNA (siRNA) in the in vitro model. Cell proliferation, migration capacity, and apoptosis in XPO5 knockdown HNSCC cell lines were evaluated by MTT, wound-healing, and caspase-3 assay, respectively. Results: Expression of XPO5 was determined to be upregulated at mRNA, protein, and intracellular level in metastatic cells compared to primary cells in HNSCC. XPO5 gene expression was knockdown by XPO5-siRNA transfection, verifying that it was suppressed at the mRNA, protein, and intracellular level. Silencing XPO5 caused a decrease in cell proliferation, delay in wound healing, and increase in Caspase-3 enzyme activity in HNSCC cell lines compared to control. Conclusions: This report is the first to describe the oncogenic role of XPO5 in HNSCC biology by in vitro experiments. Consequently, XPO5 can be used as a potential biomarker and therapeutic target molecule against the disease in the diagnosis-treatment-follow-up of HNSCC. Level of Evidence: NA Laryngoscope, 132:569–577, 2022. © 2021 The American Laryngological, Rhinological and Otological Society, Inc.Öğe Phyllanthus emblica loaded wound dressing materials(Adana Alparslan Türkeş Bilim ve Teknoloji Üniversitesi, 2023) Canatar, İpek; Özdaş, Sibel; Peşint, Gözde BaydemirAntik çağlardan beri bitkiler, çeşitli hastalık ve rahatsızlıklar için terapötik yaklaşımlarda hayati bir rol oynamıştır. Phyllanthus emblica'nın (P. emblica) birçok çalışmada çoklu terapötik aktivitelere sahip olduğu tanımlanmıştır ve Ayurvedik ve Unani tıbbi sistemlerinin önemli bir parçası olarak kabul edilir. Bu çalışma, sentezlenen P. emblica yüklü kriyojellerin yara iyileşmesi üzerindeki etkilerini ve yara pansuman uygulamalarındaki potansiyellerini incelemeyi amaçlamıştır. Bu amaçla, %0,5 (PVA/Gel/P.emblica-0,5), %1 (PVA/Gel/P.emblica-1), %1,5 (PVA/ Jel/P.emblica-1.5), %2 (PVA/Gel/P.emblica-2) P. emblica içeren polivinilalkol/jelatin (PVA/Gel) kriyojeller sentezlenmiştir. Karakterizasyon çalışmaları kapsamında şişme testleri, Brunauer-Emmett-Teller (BET) ve morfoloji analizleri yapılmıştır. Sentezlenen kriyojellerin yara pansuman malzemesi olarak uygunluğunu göstermek amacıyla HaCaT hücreleri kullanılarak in vitro hücre kültürü çalışmaları yapılmıştır. Şişme testleri ve BET analizi sonuçları, P. emblica yüklü PVA/Jel kriyojellerinin P. emblica miktarlarının artmasıyla arttığını kanıtlamıştır. Morfolojik sonuçlar, kriyojellerin yoğun, birbirine bağlı gözenek morfolojisine ve makro gözenekli bir yapıya sahip olduğunu göstermiştir. 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazolyum bromür (MTT), tripan mavisi deneyi ve canlı ölü boyama verileri, P. emblica'nın kriyojellere eklenmesinin hücre proliferasyonunun artmasına, hücre sayısının artışına ve hücre canlılığının yükselmesine yol açtığını göstermiştir. P. emblica yüklü kriyojeller üzerindeki HaCaT hücrelerinin faz kontrast mikroskop, taramalı elektron mikroskobu (SEM), immünofloresan ve giemsa görüntülerine dayanarak, P. emblica'nın hücre tutunmasını, çoğalmasını ve penetrasyonunu desteklediği gözlenmiştir. Sonuç olarak, PVA/Gel/P.emblica kriyojellerinin yara örtüsü malzemesi olarak kullanıma uygun olduğunu ve ileri çalışmalarla geliştirilebileceği gösterilmiştir.Öğe Phyllanthus emblica-Loaded Cryogels for Improved Wound Care: Characterization and In Vitro Studies(John Wiley and Sons Inc, 2024) Canatar, İpek; Özdaş, Sibel; Baydemir Peşint, GözdeWound dressings developed by combining plant extracts with polymers have made a great progress in wound care treatment. One plant with remarkable healing properties is Phyllanthus emblica Linn (P. emblica), which is described as having potent antioxidant, antimicrobial and anti-inflammatory properties. The aim of this study is to evaluate the biocompatibility of P. emblica-loaded polyvinyl alcohol/gelatin-based cryogels (PVA/Gel/P.emblica) through cytotoxicity and proliferation tests in HaCaT cells and examine their potential in wound dressing applications. Accordingly, PVA/Gel/P.emblica cryogels are successfully synthesized and characterization studies and in vitro cell culture studies are performed. The swelling tests and Brunauer–Emmett–Teller analysis results show that swelling and surface area properties of cryogels increase with increasing P. emblica amounts. Morphological results display that the cryogels have a dense, interconnected pore morphology and a macroporous structure. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, trypan blue exclusion, and live–dead assay results reveal that P. emblica enhances cell proliferation, increases cell number, and improves cell viability. Based on the scanning electron microscope, immunofluorescence, and Giemsa staining images, it is observed that P. emblica promotes cell attachment, proliferation, and penetration. These findings confirm that PVA/Gel/P.emblica cryogels are suitable for use as wound dressing materials and can be developed with further studies. © 2024 The Authors. Macromolecular Materials and Engineering published by Wiley-VCH GmbH.Öğe Protective Effect of 5-HTTLPR (S) and VNTR (10) Allele Combinationsof5-HTT Gene Against Adenotonsillary Hypertrophy(2021) Muz, Sami Engin; Özdaş, Sibel; Özdaş, Talih; Atilla, Mahmut Huntürk; Baştimur, Sibel; Öz, Işılay; Canatar, İpekINTRODUCTION: Serotonin transporter protein which is coded by 5HTT gene is responsible forpresynaptic reuptake of serotonin. In this study, we investigated the relationship between polymorphismsin the promoter region (5-HTTLPR) and in the second intron (VNTR) in the 5-HTTgene and adenotonsillarhypertrophy (ATH) in pediatric cases. METHODS: Genotyped of the 5-HTT gene promotor 5-HTTLPR and intronic VNTR of in 197 childrenwere analyzed using Snap Shot, Multifactor Dimensionality Reduction (MDR) software and carried out toassess the interactions among two polymorphisms and phenotype. RESULTS: A total of 119 children with ATH (48 girls, 71 boys age range: 3-10 years; mean age: 5.38years) and 78 healthy children (27 girls, 51 boys, age range: 4-13 years; mean age: 6.76 years) wereincluded in this study. The frequencies of the genotype in all of inheritance models of the 5-HTTLPR andthe VNTR (10) allele showed no significant differences between ATH patient and healthy controls (for allP> 0.05). However, frequency of the 5-HTTLPR (S) allele and VNTR_5-HTTLPR (10/S) haplotype and(10/10+S/S) diplotype were significantly higher in the control group compared to ATH cases (P= 0.048,P= 0.041, P= 0.13). DISCUSSION AND CONCLUSION: In this study, we observed that S/S genotype, 10/S haplotype and10/10 + S/S diplotype of 5-HTT gene could have protective effect against ATH.Öğe Pterostilbene loaded poly(vinyl alcohol)-gelatin cryogels as potential bioactive wound dressing material(John Wiley and Sons Inc, 2023) Canatar, İpek; Zenger, Okan; Özdaş, Sibel; Baydemir Peşint, GözdeCryogels are support materials which are good at mimicking extracellular matrix due to their excellent hydrophilicity, biocompatibility, and macroporous structure, thus they are useful in facilitating cell activities during healing process. In this study, polyvinyl alcohol-gelatin (PVA-Gel) based cryogel membranes loaded with pterostilbene (trans-3,5-dimethoxy-4-hydroxystilbene; PTS) (PVA-Gel/PTS) was synthesized as wound dressing materials. PVA-Gel and PVA-Gel/PTS were synthesized with the polymerization yields of 96% ± 0.23% and 98% ± 0.18%, respectively, and characterized by swelling tests, Brunauer–Emmett–Teller (BET) and scanning electron microscopy (SEM) analysis. The swelling ratios were calculated as 98.6% ± 4.93% and 102% ± 5.1%, macroporosities were determined as 85% ± 2.13% and 88% ± 2.2%, for PVA-Gel and PVA-Gel/PTS, respectively. It was determined that PVA-Gel and PVA-Gel/PTS have 17 m2/g ± 0.76 m2/g and 20 m2/g ± 0.92 m2/g surface areas, respectively. SEM studies were demonstrated that they have ~100 ?m pore sizes. According to 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), trypan blue exclusion and live-dead assay results, it was observed that cell proliferation, cell number and cell viability were higher in PVA-Gel/PTS cryogel at 24, 48, and 72 h compared to PVA-Gel. A strong and transparent fluorescent light intensity was observed indicating higher cell population in PVA-Gel/PTS in comparison with PVA-Gel, according to 4?,6-diamidino-2-phenylindole (DAPI) staining. SEM, F-Actin, Giemsa staining and inverted-phase microscope image of fibroblasts in PVA-Gel/PTS cryogels revealed that dense fibroblast proliferation and spindle-shaped morphology of cells were preserved. Moreover, DNA agarose gel data demonstrated that PVA-Gel/PTS cryogels had no effect on DNA integrity. Consequently, produced PVA-Gel/PTS cryogel can be used as wound dressing material to promote wound therapies, inducing cell viability and proliferation. © 2023 Wiley Periodicals LLC.
