Evaluation of radiosensitivity of iron-oxide-gold and iron oxide-silver core/shell nanoparticles loaded with Tumor Necrosis Factor Alpha (TNF-α) in radiation therapy of MDA-MB-231 breast cancer cell line

Abstract

Cancer is one of the leading causes of death worldwide. Among the primary reasons for the lack of treatment success and tumor recurrence is the development of resistance to therapeutic methods. Utilizing innovative and targeted approaches through the design of nanostructures that enhance drug effects and improve the impact of radiation in immuno\chemo-radiotherapy can play a valuable role in increasing the efficiency of treatment and reducing the side effects of each method. In this study, the synergistic effect of Fe3O4/Au and Fe3O4/Ag NPs loaded with Tumor Necrosis Factor alpha (TNF-α) in radiotherapy of breast cancer cell line MDA-MB-231 was investigated. Methods and Materials: Fe3O4/Au and Fe3O4/Ag core/shell NPs were synthesized at the Nutrition Research Center of Tabriz University of Medical Sciences. Dynamic Light Scattering (DLS), TEM, FT-IR, UV-Visible were used to investigate the physical and chemical characteristics of NPs. On the other hand, to check the blood compatibility of NPs, a hemolysis test was performed. TNF-α was incubated at pH=8 and temperature of 4°C with each of the nanoparticles for 24 hours to bind to the surface of the nanoparticles. TNF-α was incubated at pH=8 and temperature of 4°C with each of the NPs for 24 hours to bind to the surface. The studied cell groups included the control group and groups treated with Fe3O4/Au, TNF-α-Fe3O4/Au, Fe3O4/Ag, and TNF-α-Fe3O4/Ag, which were studied in the presence and absence of radiation. Prepared samples were transferred to the Civil Hospital for radiation exposure at doses of 2,4, and 8 Gy, using energies of 6 and 18 MV. MTT, Cell uptake, DAPI, and ROS tests were used to determine cell survival, cell uptake, apoptosis, and increased radiosensitivity, respectively. Results: The hydrodynamic diameter of Fe3O4/Au and Fe3O4/Ag NPs was 73.39 ± 2.779 and 46.91 ± 6.0682 nm, and the zeta potential was -30.6 ± 5.99 mV and 89/8 mV, respectively. Also, the TEM images of Fe3O4/Au and Fe3O4/Ag NPs showed diameters of about 24 nm and 20 nm, respectively. The correct synthesis of NPs was confirmed by FT-IR and the successful loading of TNF-α on the surface of NPs was confirmed by UV-vis. The results of the MTT assay showed that although TNF-α alone causes the proliferation of MDA-MB-231 cancer cell line, it increases cell death in the presence of radiation. Also, the anticancer effects and increased radiation sensitivity of NPs and final nano-systems were confirmed by MTT and DAPI tests in conditions with and without radiation. Both NPs showed appropriate blood compatibility up to a certain concentration. In general, cell death and apoptosis were observed following the high production of Reactive Oxygen Species (ROS) in cells treated with nano-systems + radiation.