Research On Heat Tolerance Of Blood Vessel Cells In Interventional Hyperthermia Of Liver Cancer

OBJECTIVE: Interventional hyperthermia chemotherapy is a new treatment for liver cancer,but its damage to blood vessel cells needs to be evaluated.Primary cells retained more protein structure and function than the previous used cell lines.Therefore,primary endothelial cells were extracted for in vitro hyperthermia experiments to study the changes in morphology,cell viability and death patterns in different heating conditions,and to understand the resistance of endothelial cells to hyperthermia.Then clinical strategy of hyperthermia could be put forward.Decellularization technique can preserve the intact vascular structure within the tissue,so it can be used to establish ex vitro hyperthermia models for subsequent evaluation of the effects of hyperthermia on endothelial cells.METHODS: Primary endothelial cells were isolated and cultured by tissue adherence method.The morphology changes of primary endothelial cells were studied after the treatment of water bath under temperature gradients of 41?,43? and 45?.MTT assay was used to detect the cell viability quantitatively at different temperature and heating durance,and flow cytometry was used to quantitatively detect the cell survival rate as well as apoptosis level.An ex vitro hyperthermia model was established using the decellularized liver as a biological scaffold.RESULTS: The morphology of primary endothelial cells heated at 41? for 30 min did not change significantly,while the primary endothelial cells heated at 43? for 30 min showed shrink in volume,round in shape,lose intercellular connections and accumulation of mitochondria.The morphological changes were associated with cell viability and the absolute proportion of dead cells increased.After prolonged heat treatment,the cell viability further decreased,and the activity of the primary endothelial cells continued to decrease within 24 h after heating at 45?,losing the ability to proliferate.At the end of hyperthermia,the rate of cell necrosis increased with increasing temperature.The ex vitro model of decellularized liver had a complete vascular structure with endothelial cells clustered around the vessels.CONCLUSION: Temperature plays a decisive role in the injury of hyperemia,and it hardly affects the morphology and activity of cells when the temperature is lower than the threshold temperature.Prolonged hyperthermia also increases the injury.Irreversible damage occurred to primary endothelial cells at 45?,and the cell injury was mainly necrosis at the end of warm treatment.It is suggested that the clinical treatment strategy of low temperature for a long time could be beneficial.The endothelial decellularized liver model is expected to be a good model for ex vitro interventional hyperthermia.