Nanostructured zinc carbonate hydroxide microflakes: assessing the toxicity against erythrocytes and L929 cells in vitro
Prokopiuk V. Onishchenko A. Pazura Y. Bespalova I. Kökbaş U. Tryfonyuk L. Mateychenko P. Kot K. Kurmangaliyeva S. Kot Y. Yefimova S. Tkachenko A.
24 February 2025Institute of Physics
Nanotechnology
2025#36Issue 8
Nanostructured materials have been suggested to be used as a source of dietary zinc for livestock animals. In this study, we assessed the cytotoxicity of newly synthesized nanostructured zinc carbonate hydroxide (ZnCH) Zn5(CO3)(OH)6 microflakes. Cytotoxicity of the microflakes was assessed against murine L929 cell line and rat mature erythrocytes. Viability, motility, cell death pathways, implication of Ca2+, reactive oxygen species and reactive nitrogen species (RNS) signaling, caspases, and alterations of cell membranes following exposure of L929 cells to the microflakes were assessed. To assess hemocompatibility of the Zn-containing microflakes, osmotic fragility and hemolysis assays were performed, as well as multiple eryptosis parameters were evaluated. Our findings indicate a dose-response cytotoxicity of ZnCH microflakes against L929 cells with no toxicity observed for low concentrations (10 mg l−1 and below). At high concentrations (25 mg l−1 and above), ZnCH microflakes promoted nitrosyl stress, Ca2+- and caspase-dependent apoptosis, and altered lipid order of cell membranes in a dose-dependent manner, evidenced by up to 7-fold elevation of RNS-dependent fluorescence, 2.9-fold enhancement of Fura 2-dependent fluorescence, over 20-fold elevation of caspases-dependent fluorescence (caspase-3, caspase-8, and caspase-9), and up to 4.4-fold increase in the ratiometric index of the NR12S probe. Surprisingly, toxicity to enucleated mature erythrocytes was found to be lower compared to L929 cells. ZnCH microflakes induced eryptosis associated with oxidative stress, nitrosyl stress, Ca2+ signaling and recruitment of caspases at 25-50-100 mg l−1 . Eryptosis assays were found to be more sensitive than evaluation of hemolysis. Zn5(CO3)(OH)6 microflakes show no cytotoxicity at low concentrations indicating their potential as a source of zinc for livestock animals.
cytotoxicity , eryptosis , erythrocytes , reactive oxygen species , regulated cell death
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Research Institute of Experimental and Clinical Medicine, Kharkiv National Medical University, 4 Nauky ave, Kharkiv, 61022, Ukraine
Department of Cryobiochemistry, Institute for Problems of Cryobiology and Cryomedicine, The National Academy of Sciences of Ukraine, 23 Pereyaslavskaya st, Kharkiv, 61015, Ukraine
Department of Nanostructured Materials, Institute for Scintillation Materials, The National Academy of Sciences of Ukraine, 60 Nauky ave, Kharkiv, 61072, Ukraine
Medical Biochemistry Department, Nevsehir Haci Bektas Veli University, 2000 Evler Mah. Zübeyde Hanım Cad, Nevşehir, 50300, Turkey
Institute of Health, National University of Water and Environmental Engineering, 11 Soborna st, Rivne, 33028, Ukraine
Institute for Single Crystals, National Academy of Sciences of Ukraine, 60 Nauky ave, Kharkiv, 61072, Ukraine
Department of Biochemistry, V. N. Karazin Kharkiv National University, 4 Svobody sq, Kharkiv, 61022, Ukraine
Department of Microbiology, Virology and Immunology, West Kazakhstan Marat Ospanov Medical University, 68 Maresyev st, Aktobe, 030012, Kazakhstan
BIOCEV, First Faculty of Medicine, Charles University, Průmyslová 595, Vestec, 25250, Czech Republic
Research Institute of Experimental and Clinical Medicine
Department of Cryobiochemistry
Department of Nanostructured Materials
Medical Biochemistry Department
Institute of Health
Institute for Single Crystals
Department of Biochemistry
Department of Microbiology
BIOCEV
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