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Petro Virych,1 Pavlo Virych,2 Volodymyr Prokopiuk,3,4 Anatolii Onishchenko,3 Mykola Ischenko,2 Volodymyr Doroschuk,2 Valentyna Kurovska,5 Anton Tkachenko,4 Nataliya Kutsevol2 1Laboratory of Mechanisms of Drug Resistance, R.E. Kavetsky Institute for Experimental Pathology, Oncology and Radiobiology, Kyiv, Ukraine; 2Faculty of Chemistry, Taras Shevchenko National University of Kyiv, Kyiv, Ukraine; 3Research Institute of Experimental and Clinical Medicine, Kharkiv National Medical University, Kharkiv, Ukraine; 4Department of Cryobiochemistry, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv, Ukraine; 5Educational and Scientific Center “Institute of Biology and Medicine”, Taras Shevchenko National University of Kyiv, Kyiv, UkraineCorrespondence: Pavlo Virych, Faculty of Chemistry, Taras Shevchenko National University of Kyiv, 60 Volodymyrska str, Kyiv, 01601, Ukraine, Email pavlo.virych@knu.ua Anton Tkachenko, Department of Cryobiochemistry, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, 23 Pereyaslavskaya St, Kharkiv, 61015, Ukraine, Email as.tkachenko@knmu.edu.uaIntroduction: Tumor drug resistance and systemic toxicity are major challenges of modern anticancer therapy. Nanotechnology makes it possible to create new materials with the required properties for anticancer therapy.Methods: In this research, Dextran-graft-Polyacrylamide/ZnO nanoparticles were used. The study was carried out using prostate (DU-145, LNCaP, PC-3), breast (MDA-MB-231, MCF-7, MCF-7 Dox) cancer cells and non-malignant (MAEC, BALB/3T3 clone A31) cells. Zinc was visualized with fluorescence in vitro and in vivo. ROS and apoptotic markers were identified by cytometry. Zinc accumulation and histopathological changes in the tumor, liver, kidney, and spleen were evaluated in a rat model.Results: ZnO nanoparticles dissociation and release of Zn2+ into the cytosol occurs in 2– 3 hours for cancerous and non-cancerous cells. ROS upregulation was detected in all cells. For non-malignant cells, the difference between the initial ROS level was insignificant. The rate of carbohydrate metabolism in cancer cells was reduced by nanosystems. Zinc level in the tumor was upregulated by 25% and 39% after treatment with nanosystems and doxorubicin combined, respectively. The tumor Walker-256 carcinosarcoma volume was reduced twice following mono-treatment with the nanocomplex and 65-fold lower when the nanocomplex was combined with doxorubicin compared with controls. In the liver, kidney and spleen, the zinc level increased by 10– 15% but no significant pathological alterations in the tissues were detected.Conclusion: D-PAA/ZnO NPs nanosystems were internalized by prostate, breast cancer cells and non-malignant cells via endocytosis after short time, but cytotoxicity against non-cancer cells were significantly lower in vitro and in vivo. D-PAA/ZnO NPs nanocomplex efficiently promoted cell death of tumor cells without showing cytotoxicity against non-malignant cells making it a promising anti-cancer agent.Keywords: zinc oxide nanoparticles, doxorubicin, prostate cancer, breast cancer, zinc imaging, histopathology

Vasyl Chumachenko,1 Pavlo Virych,1 Guochao Nie,2 Petro Virych,3 Oleg Yeshchenko,4 Pavlo Khort,4 Anton Tkachenko,5 Volodymyr Prokopiuk,5,6 Nataliia Lukianova,3 Taras Zadvornyi,3 Michel Rawiso,7 Liyao Ding,2 Nataliya Kutsevol1,7 1Chemistry Department, Taras Shevchenko National University of Kyiv, Kyiv, Ukraine; 2Guangxi Universities Key Laboratory of Complex System Optimization and Big Data Processing, Yulin Normal University, Yulin, People’s Republic of China; 3Laboratory of Mechanisms of Drug Resistance, R.E. Kavetsky Institute for Experimental Pathology, Oncology and Radiobiology, Kyiv, Ukraine; 4Physics Department, Taras Shevchenko National University of Kyiv, Kyiv, Ukraine; 5Research Institute of Experimental and Clinical Medicine, Kharkiv National Medical University, Kharkiv, Ukraine; 6Department of Cryobiochemistry, Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine, Kharkiv, Ukraine; 7Institut Charles Sadron, Strasbourg, FranceCorrespondence: Guochao Nie, Guangxi Universities Key Laboratory of Complex System Optimization and Big Data Processing, Yulin Normal University, Yulin, 537000, People’s Republic of China, Tel +86-17377071871, Email bccu518@163.com Pavlo Virych, Chemistry Department, Taras Shevchenko National University of Kyiv, 60 Volodymyrska str., Kyiv, 01601, Ukraine, Tel +380-631172137, Email pavlo.virych@knu.uaIntroduction: Cancer chemotherapy faces two major challenges – high toxicity of active substances and tumor resistance to drugs. Low toxic nanocarriers in combination with anticancer agents can significantly increase the effectiveness of therapy. Modern advances in nanotechnology make it easy to create materials with the necessary physical and chemical properties.Methods: Two hybrid nanosystems of dextran-polyacrylamide/ zinc oxide nanoparticles (D-PAA/ZnO NPs) were synthesized in aqueous solution with zinc sulphate (D-PAA/ZnO NPs (SO42-)) and zinc acetate (D-PAA/ZnO NPs (-OAc)). The light absorption, fluorescence, dynamic light scattering and transmission electron microscopy for nanocomposite characterization were used. MTT, neutral red uptake and scratch assays were selected as fibroblasts cytotoxicity assays. Cytotoxicity was tested in vitro for normal fibroblasts, MAEC, prostate (LNCaP, PC-3, DU-145) and breast (MDA-MB-231, MCF-7) cancer cells lines. Immunocytochemical methods were used for detection of Ki-67, p53, Bcl-2, Bax, e-cadherin, N-cadherin and CD44 expression. Acridine orange was used to detect morphological changes in cells.Results: The radius of ZnO NPs (SO42-) was 1.5 nm and ZnO NPs (-OAc) was 2 nm. The nanosystems were low-toxic to fibroblasts, MAEC. Cells in the last stages of apoptosis with the formation of apoptotic bodies were detected for all investigated cancer cell lines. Proapoptotic proteins expression in cancer cells indicates an apoptotic death. Increased expression of E-cadherin and N-cadherin was registered for cancer cells line LNCaP, PC-3, DU-145 and MCF-7 after 48 h incubation with D-PAA/ZnO NPs (SO42-).Conclusion: The nanosystems were low-toxic to fibroblasts, MAEC. The D-PAA/ZnO NPs nanosystem synthesized using zinc sulphate demonstrates high cytotoxicity due to destruction of various types of cancer cells in vitro and potentially increases adhesion between cells. Thus, our findings indicate the selective cytotoxicity of D-PAA/ZnO NPs against cancer cells and can be potentially used for cancer treatment.Keywords: zinc oxide nanoparticles, polymer nanocarrier, drug delivery, cytotoxicity, prostate cancer, breast cancer