Investigation of morphological behavior of large vessel human endothelial cells during radiation therapy based on cytoskeleton imaging

Objective: Pathological changes to endothelial cells of the vessel wall may lead to vascular stenosis. In this study, we investigate damages that appear following radiotherapy in two states, single fraction and fractionation irradiation, as an effective sign of cytoskeletal and nuclei structure of vascular wall endothelial cells. Methods: We irradiated human vein endothelial cells (HUVECs) with a Cobalt-60 therapy machine at radiation doses of 0, 2, 4, and 8 Gy. We stained the skeletal structure of the membrane and nuclei within 24 h after irradiation. This cell line received fractionation radiation therapy at doses from 0 to 8 Gy, in sub-fractions of 2 Gy, after which we stained the cytoskeleton. Morphological parameters such as area and perimeter of the cells and nuclei were determined, and we evaluated the cell shape index (CSI) for cells from each group. Results:  Increasing the irradiation dose from 0 to 8 Gy led to a significant decrease of CSI (approximately 56%) and a significant increase of nuclei shape index (approximately 85%; p<0.05). The cells’ areas and nuclei doubled (p<0.05). CSI parameter significantly decreased approximately 23% (4.5 times lower) during the radiotherapy fraction after application of the 8 Gy compared with control group cells. Conclusion: After irradiation, we observed broken the membrane filaments that resulted in a new configuration, which led to increased cell and nuclei sizes along with alterations in the cell shape. Radiation therapy led to dose-dependent changes in morphological behavior response of the endothelial cells. Hence, it would be considered as a prognostic factor for behavior of healthy vascular cells in the process of radiotherapy.

Investigation of morphological behavior of large vessel human endothelial cells during radiation therapy based on cytoskeleton imaging Keywords:

Endothelial cells , Cytoskeleton , Morphology , Radiation therapy , سلولهای اندوتلیال , اسکلت سلولی , ریختشناسی , پرتودرمانی