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    Alchemilla l. cinsine ait bazı türlerden elde edilen ekstrelerin MCF-7 kanser ve l929 fibroblast hücrelerine etkisi
    (Kırıkkale Üniversitesi, 2012) Arat, Esra; Türk, Mustafa
    Sunulan bu çalışmada, Alchemilla cinsine ait A.barbatiflora , A.tiryalensis , A.orduensis , A.speciosa türlerinin MCF-7 kanser ve L929 fibroblast hücre hatları üzerindeki etkisi araştırılmıştır.Bitki ekstraktlarının toksisite değerleri belirlenmiş,toksisitenin görüldüğü dozlarda apoptotik ve nekrotik etkiler araştırılmıştır. Sonuçların güvenilirliği açısından da xCELLigence RTCA ile hücrelerin zamana bağlı olarak değişen proliferasyon grafikleri tespit edilmiştir. Öncelikle bitki ekstraktlarının MCF-7 kanser ve L929 fibroblast hücreleri üzerindeki toksisitesi belirlenmiştir . Toksisiteyi belirlemek amacıyla WST-1 metodu kullanılmıştır. Elde edilen verilere göre A.barbatiflora'nın L929 fibroblast hücreleri üzerinde en fazla toksik etkiye sahip olduğu görülmüştür. MCF-7 kanser hücrelerinde en yüksek toksik etki ise A.tiryalensis'te gözlenmiştir. Sonuç olarak L929 fibroblast hücrelerinde ekstraktların daha fazla toksik etkiye sahip olduğu gözlenmiştir. Ekstraktların apoptotik-nekrotik indeksini belirlemek amacıyla ikili boyama metodu kullanılmıştır. İkili boyama sonuçlarına göre L929 fibroblast hücrelerinde3A.barbatiflora, diğer türlere göre en fazla apoptotik etkiye sahip türdür. MCF7 hücrelerinde ise apoptotik indeksin en fazla olduğu tür A.tiryalensis'tir. Nekrotik indeksler incelendiğinde L929 fibroblast hücresi için nekrotik indeksin en yüksek olduğu tür A.barbatiflora olarak belirlenmiştir. MCF7 hücrelerinde ise A.speciosa'nın nekrotik etkisi diğer türlere göre fazladır. Son olarak MCF-7 kanser ve L929 fibroblast hücrelerine ekstraktlar uygulandıktan sonra zamana bağlı olarak hücre proliferasyonları belirlenmiştir.Anahtar kelimeler: MCF-7, L929 fibroblast, apoptoz, nekroz, toksisite
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    Öğe
    Centaurea mersinensis phytochemical composition and multi-dimensional bioactivity properties supported by molecular modeling
    (Taylor and Francis Ltd., 2024) Yırtıcı, Ümit; Ergene, Aysun; Adem, Şevki; Atalar, Mehmet Nuri; Eyüpoğlu, Volkan; Rawat, Ravi; Arat, Esra
    Various studies conducted on Centaurea species indicate that the relevant plant is good source of bioactive phytochemicals. In this study, in vitro studies were used to determine bioactivity properties of methanol extract of Centaurea mersinensis - endemic species in Turkey - on extensive basis. Furthermore, the interaction of target molecules, identified for breast cancer and phytochemicals in the extract, was investigated via in silico analyses to support findings received in vitro. Scutellarin, quercimeritrin, chlorogenic acid and baicalin were primary phytochemicals in the extract. Methanol extract and scutellarin had higher cytotoxic effects against MCF-7 (IC50=22.17 µg/mL, and IC50=8.25 µM, respectively), compared to other breast cancer cell lines (MDA-MB-231, SKBR-3). The extract had strong antioxidant properties and inhibited target enzymes, especially ?-amylase (371.69 mg AKE/g extract). The results of molecular docking indicate that main compounds of extract show high-strength bonding to the c-Kit tyrosine among target molecules identified in breast cancer, compared to other target molecules (MMP-2, MMP-9, VEGFR2 kinase, Aurora-A kinase, HER2). The tyrosinase kinase (1T46)-Scutellarin complex showed considerable stability in 150 ns simulation as per MD findings, and it was coherent with optimal docking findings. Docking findings and HOMO-LUMO analysis results corresponds with in vitro experiments. Medicinal properties of phytochemicals, which was determined to be suitable for oral use along with ADMET, were found to be within normal limits except for their polarity properties. In conclusion, in vitro and in silico studies indicated that the relevant plant yields promising results regarding its potential to develop novel and effective medicational products. Communicated by Ramaswamy H. Sarma. © 2023 Informa UK Limited, trading as Taylor & Francis Group.
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    Centaurea mersinensis phytochemical composition and multi-dimensional bioactivity properties supported by molecular modeling
    (Taylor & Francis Inc, 2024) Yirtici, Umit; Ergene, Aysun; Adem, Sevki; Atalar, Mehmet Nuri; Eyupoglu, Volkan; Rawat, Ravi; Arat, Esra
    Various studies conducted on Centaurea species indicate that the relevant plant is good source of bioactive phytochemicals. In this study, in vitro studies were used to determine bioactivity properties of methanol extract of Centaurea mersinensis - endemic species in Turkey - on extensive basis. Furthermore, the interaction of target molecules, identified for breast cancer and phytochemicals in the extract, was investigated via in silico analyses to support findings received in vitro. Scutellarin, quercimeritrin, chlorogenic acid and baicalin were primary phytochemicals in the extract. Methanol extract and scutellarin had higher cytotoxic effects against MCF-7 (IC50=22.17 mu g/mL, and IC50=8.25 mu M, respectively), compared to other breast cancer cell lines (MDA-MB-231, SKBR-3). The extract had strong antioxidant properties and inhibited target enzymes, especially alpha-amylase (371.69 mg AKE/g extract). The results of molecular docking indicate that main compounds of extract show high-strength bonding to the c-Kit tyrosine among target molecules identified in breast cancer, compared to other target molecules (MMP-2, MMP-9, VEGFR2 kinase, Aurora-A kinase, HER2). The tyrosinase kinase (1T46)-Scutellarin complex showed considerable stability in 150 ns simulation as per MD findings, and it was coherent with optimal docking findings. Docking findings and HOMO-LUMO analysis results corresponds with in vitro experiments. Medicinal properties of phytochemicals, which was determined to be suitable for oral use along with ADMET, were found to be within normal limits except for their polarity properties. In conclusion, in vitro and in silico studies indicated that the relevant plant yields promising results regarding its potential to develop novel and effective medicational products.Communicated by Ramaswamy H. Sarma
  • Küçük Resim
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    Does penile tourniquet application alter bacterial adhesion to rat urethral cells: an in vitro study
    (W B Saunders Co-Elsevier Inc, 2018) Boybeyi-Turer, Ozlem; Kacmaz, Birgul; Arat, Esra; Atasoy, Pinar; Kisa, Ucler; Gunal, Yasemin Dere; Soyer, Tutku
    Purpose: To investigate the effects of penile tourniquet (PT) application on bacterial adhesion to urothelium. Methods: Fifty-six ratswere allocated into control group (CG), shamgroup (SG), PT group (PTG). No intervention was applied in CG. A 5mm-length urethral repair was performed in SG and PTG. In PTG, a 10-min duration of PT was applied during the procedure and the tissue oxygenation monitor was used to adjust the same degree of ischemia in all subjects. Sampleswere examined for wound healing parameters and tissue levels of inflammatory markers, eNOS, e-selectin, and ICAM-1antibodies. The adhesion of Escherichia coli to urotheliumwas investigated with in vitro adhesion assay. Results: Inflammation was higher and wound healing was worse in SG than CG and in PTG in comparison to CG and SG (p < 0.05). The endothelial damage, as shown by eNOS expression, was significantly higher in PTG compared to CG and SG (p < 0.05). The staining with ICAM-1 and e-selectin antibodies, showing increased inflammatory response to bacterial adhesion, was significantly higher in PTG compared to CG and SG (p < 0.05). In vitro urethral cell proliferation was achieved only in CG and SG revealing significantly increased adhesion in SG compared to CG (p < 0.05). The PT application caused endothelial corruption and prevented cell proliferation in cell culture. Conclusion: The PT application does not improve wound healing and increases bacterial adhesion molecules in penile tissue. The in vitro assays showed that PT causes severe endothelial damage and inhibits endothelial cell proliferation. (c) 2017 Elsevier Inc. All rights reserved.
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    Doku rejenerasyonunu, ekstraselüler matriks sentezini artıran kolay uygulanabilir bitkisel kaynaklı ürün geliştirilmesi
    (Kırıkkale Üniversitesi, 2019) Arat, Esra; Türk, Mustafa
    Çalışmamızda C.coggyria (Duman ağacı), M.communis (Mersin bitkisi), T.spicata (Karabaş kekik) bitkisine ait metanol ve saf su ekstrelerinin in vitro çalışmalarla L929 fibroblast hücreleri üzerindeki etkisi araştırılmıştır. Hücre morfolojisinin incelenmesi için hematoksilen eozin boyama yapılmıştır. Ekstrelerin sitotoksisitesinin belirlenmesi amacıyla WST-1 testi kullanılmıştır. Apoptoz ve nekrozun belirlenmesi için double staining-ikili boyama yöntemi kullanılmış ve WST-1 ile apoptoz ve nekrozun belirlendiği konsantrasyonlara bağlı olarak xCELLigence-Gerçek Zamanlı Hücre Analiz Sistemi'nde bitkilere ait su ekstreleri uygulanarak hücre proliferasyonu belirlenmiştir. Buna ek olarak mikronükleus testi ve Salmonella AMES testi ile bitki ekstrelerinin genotoksik ve mutajenik etkisinin olup olmadığı araştırılmıştır. Yara iyileşmesinde önemli rol oynayan kollajen tip I, tip IV, fibronektin ve elastin protein ekspresyonları western blot, immunositokimyasal ve immunohistokimyasal yöntemlerle belirlenmiştir. In vitro deney sonuçlarına bağlı olarak in vivo deneylerde kullanılan ekstrelerin konsantrasyonları belirlenmiştir. In vivo çalışmalarla bitki ekstrelerinin deri irritasyon, göz irritasyon, sensitizasyon, akut sistemik toksisite, yara modellerinin incelenmesi, yanık modellerinde histopatolojik incelenmesi, klinik biyokimya ve hematoloji ölçümlerine bağlı değerlendirmeler yapılmıştır. Agregasyon ve hemoliz testi ile bitki ekstrelerinin kan ile etkileşimi araştırılmıştır. In vitro deneyler sonucunda C.coggyria (Duman ağacı) bitkisine ait su ve metanol ekstrelerinin M.communis (Mersin bitkisi) ve T.spicata (Karabaş kekik) bitkilerine ait ekstrelere göre toksik etkisinin olmadığı, hücre proliferasyonunu artırdığı belirlendi. Ayrıca C.coggyria (Duman ağacı), M.communis (Mersin bitkisi) ve T.spicata (Karabaş kekik) bitkilerine ait su ekstrelerinin genotoksik etkisinin olmadığı belirlendi. In vitro deneyler sonucunda C.coggyria (Duman ağacı) bitkisine ait su ekstresinin in vivo deneylerde daha fazla iyileşme gösterdiği belirlendi. Kan ile etkileşim deneylerinden agregasyon testi sonuçlarına göre; C.coggyria (Duman ağacı), M.communis (Mersin bitkisi), T.spicata (Karabaş kekik) bitkilerine ait su ekstrelerinin agregasyona neden olmadığı, T.spicata (Karabaş kekik) bitkisine ait su ekstresinin hafif hemolitik etkisinin olduğu belirlendi. İn vitro ve in vivo yapılan deneyler sonucunda özütler arasında en iyi etki gösteren C.coggyria (Duman ağacı) bitkisinden yara yanık iyileşmesinde kullanılmak üzere ürün geliştirilmiştir.
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    Evaluation of melamine and cyanuric acid cytotoxicity: an in vitro study on L929 fibroblasts and CHO cell line
    (ANKARA UNIV PRESS, 2020) Melekoglu, Abdullah; Ekici, Husamettin; Arat, Esra; Karahan, Siyami
    Melamine and its metabolites pose health concern as they are used in various industrial products including feed and drugs. There are a limited number of studies on melamine and cyanuric acid cytotoxicity and cellular damage without a certain conclusion. The present study aimed to evaluate melamine, cyanuric acid and its combined cytotoxic effects using 3-(4.5-dimethylthiazol-2-yl) methyl thiazole tetrazolium (MTT) bromide test. The study also evaluated apoptotic and necrotic effect using a double staining method of Hoechst 33342 and propidium iodide. Melamine, cyanuric acid and their combination (1:1) were applied to L929 fibroblasts and Chinese hamster ovary (CHO) cells at various concentrations (1000 mu g/mL, 500 mu g/mL, 250 mu g/mL, 125 mu g/mL and 62.5 mu g/mL). At the highest concentration (1000 mu g/mL), the cell viability dropped down approximately to 50% both in CHO cells and L929 cells. Melamine, cyanuric acid and their mixture caused cytotoxicity in CHO cells and L929 fibroblasts in dose-dependent manner Cell death occurred through both apoptosis and mainly necrosis. Both cell types were more sensitive to the mixture of melamine and cyanuric acid and, furthermore, CHO cells were more sensitive than L929 fibroblasts. As a result, melamine, cyanuric acid and their combination caused cytotoxicity in CHO cells and L929 fibroblasts. Further studies should be conducted in different cell lines. These studies should also aim to reveal the mechanism of cytotoxicity and related pathways.
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    Investigation of the in vitro antibacterial, cytotoxic and in vivo analgesic effects of silver nanoparticles coated with Centella asiatica plant extract
    (Ankara Univ, 2023) Bozkaya, Ogun; Ekici, Husamettin; Gun Gok, Zehra; Arat, Esra; Ekici, Seda; Yigitoglu, Mustafa; Vargel, Ibrahim
    In recent years, researchers have shown an increased interest in using medicinal plants for the synthesis of silver nanoparticles (AgNPs) having various therapeutic properties. Centella asiatica (CA), a medicinal plant, has been used to treat minor burn wounds, psoriasis, and hypertrophic wounds among many other pathological conditions. The current study aimed to synthesize CA coated AgNPs (CA-AgNPs) with appropriate biocompatibility and various therapeutic properties, including antimicrobial and analgesic activities. The synthesized CA-AgNPs were characterized by ultraviolet-visible (UV-Vis) spectroscopy, zeta potential measurements, and fourier transform infrared (FT-IR) spectroscopy. The formation of spherical CA-AgNPs was confirmed by a single surface plasmon resonance (SPR) peak emerging at 420 nm wavelength by UV-Vis. The average hydrodynamic diameter and zeta potential of the particles were found to be 29.5 nm and -24.5 mV, respectively. The FT-IR analyses showed that the AgNPs were coated and stabilized by bioactive compounds from the CA extract. MTT cytotoxicity assay revealed that CA-AgNPs at <= 1 mM concentrations exhibited biocompatibility for L929 fibroblast cells. The antimicrobial activity of CA-AgNPs was confirmed by significant inhibition of Staphylococcus aureus and Escherichia coli. In addition, the analgesic effect of CA-AgNPs was investigated for the first time in the literature by tail-flick and hot plate methods, and statistically significant results were obtained for both methods. Taken together, these results suggest that CA-AgNPs can be used as an effective antibacterial and analgesic agent in a variety of biomedical applications, including coating wound dressings.
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    Production and characterization of hybrid nanofiber wound dressing containing Centella asiatica coated silver nanoparticles by mutual electrospinning method
    (Pergamon-Elsevier Science Ltd, 2022) Bozkaya, Ogun; Arat, Esra; Gok, Zehra Gun; Yigitoglu, Mustafa; Vargel, Ibrahim
    The aim of this work is to produce a therapeutic and antimicrobial nanofiber wound dressing material with suitable air permeability, water vapor transmission, water absorption, porosity, thermal and mechanical strength. For this purpose, first of all, Centella asiatica (CA) coated silver nanoparticles (CA-AgNPs) are synthesized with using CA extract as reducing and stabilizing agent. The green synthesized CA-AgNPs are characterized by UV-Vis spectroscopy, transmission electron microscope (TEM), zeta potential and fourier transform infrared (FTIR) spectroscopy measurements. The obtained CA-AgNPs give a single peak in the 420-430 nm range between 200 and 700 nm due to surface plasmon resonance (SPR). The average zeta potential and size of CA-AgNPs is found to be-30.4 mV and 14.8 +/- 7.3 nm, respectively. Then, poly caprolactone (PCL) and polyethylene oxide (PEO) nanofibers composition containing CA-AgNPs is synthesized by mutual electrospinning method. Before electrospinning, the PCL solution (12% w/v in chloroform/methanol (70%/30%, v/v)), PEO solution (3.5%, w/v in water) and PEO solutions containing 1%, 5% and 10% (v:v) of CA-AgNPs are prepared. In electrospinning experiments, to synthesis the PCL/PEO nanofibers containing CA-AgNPs, an electrospinning set-up consisting of two high voltage sources, an aluminum rotary roller collector and two syringe pumps (one with a double syringe and the other with a single syringe) is used. To produce the wound dressing materials, PEO solutions containing CA-AgNPs are placed on the double syringe pump and PCL solution is placed on the single syringe pump. The surface and physico-chemical properties of the produced hybrid nanofibers are characterized by field emission scanning electron microscopy (FESEM), energy dispersion spectrometry (EDS), FTIR, X-ray diffractometer (XRD), Brunauer-Emmett-Teller (BET) surface area and porosity analyzer. Also, the thermal and mechanical properties of the obtained materials are investigated. In addition, the air permeability, water uptake capacity, water contact angle, water vapor transmission, in vitro degradation and silver release behavior of the samples are investigated. The results show that mutual dual-spinneret electrospinning technique combines the features of dissimilar components without corruption. Moreover, the in vitro degradation profile and silver release results show that these nanofibers could be used in wound dressing applications in the long term. With antimicrobial studies, PEO/ PCL nanofibers containing 5% and 10% CA-AgNPs are found to be effective against Staphylococcus aureus, Escherichia coli and Candida albicans. Also, the cytotoxic properties of nanofibers are investigated by MTT (3-[4,5dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay and results show the good biocompatibility for L929 fibroblast cells. Results reveal that CA-AgNPs loaded PCL/PEO hybrid nanofibers materials synthesized in this study has a promising potential for wound healing applications.
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    Proliferation Of Mc3t3-E1 Osteoblastic Cells After Sequential Release Of Pdgf And Bmp-6. An In Vitro Study
    (2017) Keçeli, H. Gencay; Bayram, Cem; Ercan, Nuray; Türk, Mustafa; Arat, Esra; Nohutçu, Rahime M.
    Background and Aim: To evaluate the release kinetics of bone morphogenetic protein (BMP)-6 or -7 loaded nanoparticles (NPs) that located inside the microparticles (MPs) carrying platelet-derived growth factor (PDGF), and test this NP-in-MP system with MC3T3-E1 osteoblastic cells. Materials and Methods: Poly-lactic acid-co-glycolic acid (PLGA) NPs was loaded with BMP-6 or -7 and inserted in sodium alginate MPs loaded with PDGF. To evaluate the osteoblastic effect; proliferation of MC3T3-E1 cells that were treated with BMP-6, -7 and PDGF free solutions (FS) or within the particles (BMP-6 or -7 loaded PLGA NP alone and BMP-6 or -7 loaded PLGA NP in PDGF loaded alginate MP) were assessed at 2, 4, 7, 14 and 21 days. Results: It was shown that while both NP and NP-in-MP systems showed similar burst release at first time periods; especially in 24-72 h time period, NP-in-MP system exhibited a sustained release profile till 14th day. According to proliferation experiments, till the 7th day, both particle and FS groups exhibited similar profiles, but after that time particle groups, especially BMP-7 NP in PDGF MP, reached to statistically higher cell numbers than FS groups. NP-in-MP system exhibited a gradually longer time factor release resulting with delayed but elongated cell proliferation period. Conclusion: Findings indicate that NP-in-MP system might be promising in future for mimicking the natural bone formation process by providing sequential release of PDGF and BMPs, for bone tissue engineering. More comprehensive experiments evaluating mineralization and gene expression profile is necessary to verify these results
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    Synthesis and biological evaluation of novel urea, thiourea and squaramide diastereomers possessing sugar backbone
    (ELSEVIER SCI LTD, 2020) Isilar, Ozer; Bulut, Adnan; Yaglioglu, Ayse Sahin; Demirtas, Ibrahim; Arat, Esra; Turk, Mustafa
    A series of novel chiral 14 urea, thiourea and squaramide stereoisomers possessing carbohydrate backbones as well as amide functional groups was synthesized and characterized by their, H-1 NMR, C-13 NMR, FT-IR, HRMS, optical rotation, and melting points. Their antiproliferative activities were investigated against HeLa and PC3 cell lines. The compounds 9, 11 and 12 showed better activities at 25 mu M against PC3 cell line with respect to the standard 5-fluorouracil (5-FU). Especially, the compounds 9 and 11 showed higher activities than the standard 5-FU even at low concentration (5 mu M) against HeLa cell line. IC50 results also confirm these activities. The compounds 9, 10 and 11 have the IC50 values of 1.10 mu M, 1.51 mu M and 1.02 mu M, respectively while 5-FU has 2.51 mu M. Moreover, their cytotoxicity tests have proven that their viabilities were in between 50% and 100%.
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    Tildipirosinin Sığır Trakea Epitel Hücresi Üzerinde Yara İyileştirici Etkisi
    (2021) Şahin, Yaşar; Yıldırım, Ebru; Ekici, Hüsamettin; Türk, Mustafa; Arat, Esra
    Bu çalışmada; sığır solunum yolu hastalıkları tedavisinde kullanılan tildipirosinin, sığır trakea epitel hücresi üzerinde invitro yara iyileşmesi etkisinin araştırılması amaçlandı. In vitro yara yöntemi ile her kuyucuğa ekilmiş olan sığır trakea epitel hücreleri üzerinde, 200 ?L’lik pipet ucu (dikey) aracılığıyla bir yara alanı oluşturuldu. In vitro yara oluşturulmuş sığır trakeaepitel hücreleri üzerine tildipirosinin konsantrasyonları (5 ?g/mL, 10 ?g/mL, 25 ?g/mL, 50 ?g/mL) ve kontrol grubuuygulandı. In vitro yara görüntüleri; 0’ıncı saat, 24’üncü saat ve 48’inci saatte izlendi. Çalışmanın 24’üncü saatinde yara alanıkapanma yüzdesi, kontrol grubuna kıyasla; tildipirosinin 50 ?g/mL konsantrasyonda istatistiki olarak önemli farkbulunamazken 5, 10 ve 25 ?g/mL konsantrasyonlarında istatistiki olarak fark anlamlı bulundu. Çalışmanın 48’inci saatinde,kontrol grubuna kıyasla 25 ve 50 ?g/mL tildipirosin konsantrasyonları arasında istatistiki olarak önemli fark bulunamazken, 5?g/mL (%96,61±1,69) ve 10 ?g/mL (%96,42±1,78) tildipirosin konsantrasyonları arasında yara alanı kapanma yüzdeleriarasında istatistiki olarak fark anlamlı bulundu. Sonuç olarak tildipirosinin düşük konsantrasyonlarındaki yara iyileşmeyüzdelerinin (48’inci saat) yaklaşık olarak %96 olması, hastalıkların tedavisine ek olarak çeşitli mikroorganizma ve maddelertarafından epitel hücrelerinde meydana gelen hasarlara karşı doku bütünlüğünü koruma ve tedaviye katkı sağlayabilir.