Browsing by Author "Ensarioğlu H.K."

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    Boron-Doped Carbon Nanodots as a Theranostic Agent for Colon Cancer Stem Cells
    (American Chemical Society, 2023) Ozkasapoglu S.; Caglayan M.G.; Akkurt F.; Ensarioğlu H.K.; Vatansever H.S.; Celikkan H.
    Carbon nanodots have drawn a great deal of attention due to their green and expedient opportunities in biological and chemical sciences. Their high fluorescence capabilities and low toxicity for living cells and tissues make them excellent imaging agents. In addition, they have a fluorimetric response against inorganic and organic species. Boron-doped carbon nanodots (B-CDs) with high fluorescence yield were produced from phenylboronic acid and glutamine as boron and carbon sources, respectively, by a hydrothermal method. First, the effects of the temperature on their fluorescence yield and the structural characteristics of B-CDs were investigated. Second, their cytotoxicity and cell death and proliferation behaviors were examined. The cytotoxicity was evaluated by the MTT assay. The cellular properties were evaluated with the distribution of caspase 3, Ki67, lamin B1, P16, and cytochrome c after the indirect immunoperoxidase technique. After the MTT assay, 1:1 dilution of all applicants for 24 h was used in the study. After immunohistochemical analyses, the application of B-CDs synthesized at 230 °C did not change control cell (Vero) proliferation, and also apoptosis was not triggered. Colo 320 CD133+ and CD133- cell-triggered apoptosis and cellular senescence were found to be synthesis temperature dependent. In addition, Colo 320 CD133- cells were affected relatively more than CD133+ cells from B-CDs. While B-CDs did not affect the control cells, the colon cancer stem cells (Colo 320 CD133+) were affected in a time-dependent manner. Therefore, the use of the synthesized B-CD product may be an alternative method for controlling or eliminating cancer stem cells in the tumor tissue. © 2023 The Authors. Published by American Chemical Society
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    Cell Death Mechanism of Organometallic Ruthenium(II) and Iridium(III) Arene Complexes on HepG2 and Vero Cells
    (American Chemical Society, 2023) Kavukcu S.B.; Ensarioğlu H.K.; Karabıyık H.; Vatansever H.S.; Türkmen H.
    Due to side effects and toxicity associated with platinum-derived metal-based drugs, extensive research has been conducted on ruthenium (Ru) complexes. We aim to synthesize a highly oil soluble Ru(II)-p-cymene complex (Ru1) with an aliphatic chain group, a bimetallic Ru(II)-p-cymene complex (Ru2) with N,S,S triple-coordination and a bimetallic Ir(III)-pentamethylcyclopentadienyl complex (Ir1) with S,S double-coordination. Subsequently, we investigate the effects of these complexes on Vero and HepG2 cell lines, focusing on cell death mechanisms. Characterization of the complexes is performed through nuclear magnetic resonance spectroscopy (1H and 13C NMR) and Fourier-transform infrared spectroscopy. The effective doses are determined using the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) (MTT) assay, applying different doses of the complexes to the two cell lines for 24 and 48 h, respectively. Immunoreactivities of Bax, Bcl2, caspase-3, RIP3, and RIPK1 are analyzed using the indirect immunoperoxidase technique. Notably, all the complexes (Ru1, Ru2, and Ir1) exhibit distinct cell death mechanisms, showing greater effectiveness than cisplatin. This study reveals the diverse mechanisms of action of Ru and Ir complexes based on different ligands. To the best of our knowledge, this study represents the first investigation of a novel RAED-type complex (Ru1) and unexpected bimetallic complexes (Ru2 and Ir1). © 2023 The Authors. Published by American Chemical Society.
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    A newly synthesized thiosemicarbazide derivative trigger apoptosis rather than necroptosis on HEPG2 cell line
    (John Wiley and Sons Inc, 2024) Başoğlu-Ünal F.; Becer E.; Ensarioğlu H.K.; -Güzeldemirci N.U.; Kuran E.D.; Vatansever H.S.
    Thiosemicarbazide derivatives have been the focus of scientists owing to their broad biological activities such as anticancer, antimicrobial, and anti-inflammatory. Herein, we designed and synthesized a new thiosemicarbazide derivative (TS-1) and evaluated its antiproliferative potential against the human hepatocellular carcinoma cell line (HEPG2) and human umbilical vein endothelial cell line (ECV-304). Also, it was aimed to investigate the necroptotic and apoptotic cell death effects of TS-1 in HEPG2 cells, and these effects were supported by molecular docking. The new synthesized compound structure was characterized using various spectroscopic methods such as FT-IR, 1H-NMR, 13C-NMR, and elemental analysis. The cytotoxic activity of the tested compound was measured by the MTT assay. Apoptotic and necroptotic properties of the TS-1 were evaluated by indirect immunoperoxidase method using antibodies against Ki-67, Bax, Bcl-2, caspase-3, caspase-8, caspase-9, RIP3, and RIPK1. Apoptotic and necroptotic effects of TS-1 were supported by molecular docking. Compound TS-1 was synthesized as a pure compound with a high yield. The effective value of TS-1 was 10 μM in HEPG2 cells. TS-1 did not show any cytotoxic effect on ECV-304. Caspase-3 and RIPK1 immunoreactivities were significantly increased in HEPG2 cells after being treated with TS-1. As the results of the molecular docking studies, the molecular docking showed that the TS-1 exhibits H-bond interaction with various significant amino acid residues in the active site of both RIPK1. It could be concluded that TS-1 could be a promising novel therapeutic agent by inducing apoptosis rather than necroptosis in HEPG2 cells. © 2023 John Wiley & Sons Ltd.
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    Electrochemically generated paper SERS substrate for detection of exosome in urine samples
    (Elsevier B.V., 2025) Kayiş E.Ç.; Torul H.; Sazaklıoğlu S.A.; Çelikkan H.; Ensarioğlu H.K.; Gumus B.H.; Vatansever H.S.; Tamer U.
    Here, a single-drop paper-based surface-enhanced Raman spectroscopy (SERS) immunoassay was developed to pave the way for monitoring exosome numbers for the early diagnosis of prostate cancer. Exosomes are nano-sized (40–150 nm) membrane vesicles that provide intercellular communication. In our work, we offer a new paper SERS substrate for exosome detection in urine. We initially electrochemically deposited nanostar-shaped gold nanoparticles (AuNPs) on the working electrode to crate the paper SERS substrates. Then we functionalized them with 11-mercaptoundecanoic acid (11-MUA) and conjugated them with anti-CD9 antibodies. After capturing exosomes, the sandwich immunoassay structure was created by using gold nanorods (AuNRs) modified with 5,5-dithiobis (2-nitrobenzoic acid) (DTNB) as a Raman tag. The SERS signal intensities of DTNB molecules at 1330 cm−1 were monitored to determine the exosome concentration. Each step occurred in only one drop of solution or sample. The developed single-drop paper-based SERS immunoassay exhibited a linear range from 1.0 × 103 to 1.0 × 109 exosome particles/mL with correlation coefficients (R2) of 0.9903. The limit of detection (LOD) was found as 9.9 × 101 exosome particles/mL. The developed system was tested with clinical urine samples from patients with benign prostatic hyperplasia, prostatitis, prostate cancer, and healthy individuals. The obtained results were compared with the exosome particle numbers in these samples determined by an enzyme-linked immunosorbent assay (ELISA) method and the accuracy of the system was evaluated with an average recovery value of 96.7 %. The developed biosensor system enables highly sensitive detection of exosomes in low-volume urine samples. The usage of a paper membrane as a SERS substrate, combined with the electrochemical deposition of gold nanoparticles, provides an eco-friendly and cost-effective solution, enabling wider use and applications. © 2024 Elsevier B.V.