Accumulating evidence suggests that manganese oxide nanoparticles (NPs) show multiple enzyme-mimicking antioxidant activities, which supports their potential in redox-targeting therapeutic strategies for diseases with impaired redox signaling. However, the systemic administration of any NP requires thorough hemocompatibility testing. In this study, we assessed the hemocompatibility of synthesized Mn3O4 NPs, identifying their ability to induce spontaneous hemolysis and eryptosis or impair osmotic fragility. Concentrations of up to 20 mg/L were found to be safe for erythrocytes. Eryptosis assays were shown to be more sensitive than hemolysis and osmotic fragility as markers of hemocompatibility for Mn3O4 NP testing. Flow cytometry- and confocal microscopy-based studies revealed that eryptosis induced by Mn3O4 NPs was accompanied by Ca2+ overload, altered redox homeostasis verified by enhanced intracellular reactive oxygen species (ROS) and reactive nitrogen species (RNS), and a decrease in the lipid order of cell membranes. Furthermore, Mn3O4 NP-induced eryptosis was calpain- and caspase-dependent.

1st Department of Internal Medicine Hematology 1st Faculty of Medicine Charles University 12808 Prague Czech Republic

BIOCEV 1st Faculty of Medicine Charles University Průmyslová 595 25250 Vestec Czech Republic

Department of Biochemistry 5 N Karazin Kharkiv National 4 Svobody sq 61022 Kharkiv Ukraine

Department of Biomedical Engineering Kharkiv National University of Radio Electronics 14 Nauky ave 61116 Kharkiv Ukraine

Department of Chemistry Stockholm University Svante Arrhenius väg 16C SE 106 91 Stockholm Sweden

Department of Cryobiochemistry Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine 23 Pereyaslavskaya st 61015 Kharkiv Ukraine

Department of Microbiology Virology and Immunology West Kazakhstan Marat Ospanov Medical University 68 Maresyev st Aktobe 030012 Kazakhstan

Department of Nanostructured Materials Institute for Scintillation Materials of the National Academy of Sciences of Ukraine 60 Nauky ave 61072 Kharkiv Ukraine

Institute of Health National University of Water and Environmental Engineering 11 Soborna st 33028 Rivne Ukraine

Research Institute of Experimental and Clinical Medicine Kharkiv National Medical University 4 Nauky ave 61022 Kharkiv Ukraine

Universal Scientific Education and Research Network 61022 Kharkiv Ukraine

Ye O Paton Institute of Materials Science and Welding National Technical University of Ukraine Igor Sikorsky Kyiv Polytechnic Institute 37 Beresteiskyi ave 03056 Kyiv Ukraine

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