The effect of free and liposomal doxorubicin on the morphofunctional characteristics of saos-2 osteosarcoma cells under the influence of induction moderate hyperthermia

Orel V.Е.1, Diedkov A.G.1, Ostafiichuk V.V.1, Kolesnyk D.L.2, Dasyukevich O.J.1, Rykhalskyi О.Yu.1

Summary. Abstract. This work aims to investigate the influence of free doxorubicin (FDR) and liposomal doxorubicin (LDR) in combination with induction moderate hyperthermia (IMH) on the functional and morphological changes in Saos-2 cells in vitro. Object and methods. Human osteosarcoma Saos-2 (ATCC HTB-85) cells obtained from the bank of human and animal tissues of the R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology of the National Academy of Sciences of Ukraine were used in this study. Saos-2 cells were exposed to IMH at 42 MHz frequency of electromagnetic irradiation and 15 W power for 30 minutes. Morphofunctional changes in Saos-2 cells in response to FDR and LDR combined with IMH were examined by the flow cytometry method based on quantitative changes in indicators of the forward light scattering by cells (FSC-A) and side light scattering (SSC-A). The gating strategy for the cytometric studies was to isolate a population of minimally damaged cells and to filter out cellular debris on FSC-A/SSC-A dot plots and to exclude doublets from FSC-A/FSC-H dot plot analysis. Cytometry data analysis was performed using the CytExpert for DxFlex software. Results. FDR caused a 30% increase in the size of Saos-2 cells whereas LDR increased the cell size by 26% as compared with the control group. IMH alone initiated a minor but statistically significant increase of the cell size (1.5%, р <0.05) in comparison with the control. The combination of FDR and IMH resulted in 8% greater size of Saos-2 cells than FDR alone and 39% greater size than IMH alone (р <0.05). The combination of LDR and IMH induced a 24% increase in cell size compared to IMH alone (р <0.05). Granularity analysis revealed a 103% (р <0.05) and 97% (р <0.05) increase in response to FDR and LDR as compared with the control group, respectively. IMH alone increased cell granularity by 21.4% (р <0.05) in comparison with the control. The combined action of IMH with FDR or LDR initiated minor increase of Saos-2 cell granularity. Conclusion. It was found that the combined action of IMH and FDR had a more pronounced effect on increasing the size and granularity of Saos-2 cells than the effect of FDR. LDR alone and in combination with IMH initiated a less pronounced increase in the size and granularity of Saos-2 cells than FDR alone or in combination with IMH, which is probably due to greater cytotoxic effect of FDR. IMH also contributes to an increase in the size and granularity of Saos-2 cells when compared to the control group.

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