Investigation of drug-induced liver endothelial damage by using a liver cancer-on-chip platform

Objective: Today, with recent advances in microfluidics for morphological and functional reconstruction of diseased tissues, vascular components along with vascular cells have been incorporated into engineered in vitro systems. In this way, it allows more realistic modeling of the healthy or diseased 3D liver microenvironment, such as hepatocytes, which are the parenchymal cells of the liver, through the microfluidic perfusion channel of hemodynamics and drugs/soluble factor (Lauschke, 2019). Although chemotherapeutic drugs are effective on liver cancer, they are not selective against cancer cells and it can also affect healthy cells and cause toxicity. İn our study, we aimed to examine the effect of the selected chemotherapy drug (cyclophosphamide) used in liver cancer on endothelial cells by creating a 3D liver cancer model. Methods: To determine the toxic effect of the drug after exposing endothelial cells in static culture and in the chip platform for 24 and 96 hours, viability analysis with Live/Dead staining, expression of tissue factor (CD142), which is a damage marker, by flow cytometry analysis, viability and proliferation by MTT method, and morphological changes were examined by immunofluorescent labeling of the surface marker specific to endothelial cells CD31 /PECAM1. Results: It was observed that cyclophosphamide reduced the viability of both static and dynamic cultured endothelial cells. It was seen by IF labeling and Live/Dead staining analyzes that the co-cultured experimental groups were more resistant to the effect of the drug. Conclusions: In this thesis, it was concluded that CD31 and CD142 markers can help determine the damage caused by the drug cyclophosphamide on endothelial cells.