The present study aimed to assess the effects of extremely low-frequency electromagnetic fields (ELF-MF) on structural changes of human osteosarcoma cells by analyzing the stained cytoskeleton for assessing the relationship between the fractal dimension parameter and proliferation rate of radiation-induced cells. In this study, 2-mT magnetic fields with various waveforms, including sinusoidal, triangular, and pulsed shapes, were employed to determine the biological effects of ELF-EMF on the human osteosarcoma MG-63 cell line. All experiments were performed in two modes: continuous exposure at 3 h and fractionated irradiations at 3 consecutive days. Afterward, the proliferation assay was implemented for assessing the cell proliferation in each group. Moreover, immunofluorescence staining and confocal imaging were performed to determine the cell shape index. Furthermore, fractal dimension analysis was carried out by processing morphological images. The proliferation and shape index parameters of radiation-induced osteosarcomas significantly decreased compared with non-irradiated cells. In addition, fractal dimensions significantly increased following fractionated exposure at 3 consecutive days. Assessing the fractal dimensions can be considered as a new morphological index for the prognosis of the structural remodeling of human osteosarcoma cells in response to fractionated irradiation of ELF-MF. In addition, various waveforms induce a similar effect on morphological remodeling and cell proliferation.
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