Pharmacodynamic Study Of Sorafenib And Nivolumab On Liver Cancer In A Simulated Tumor Microenvironment In Vitr

This study aims to utilize 3D bioprinting technology to construct in vitro models of liver cancer,with the goal of better simulating the tumor microenvironment and evaluating the pharmacological effects of the targeted drug sorafenib and the immune checkpoint inhibitor nivolumab.We successfully applied 3D bioprinting technology to print models of two liver cancer cell lines,HepG2 and Huh7.These models possess three-dimensional structures and exhibit cellcell interactions,closely resembling the physiological conditions of real tumors.By optimizing printing parameters and cell suspension formulations,we obtained models with high cell viability and uniform cell distribution.The models were then used for drug treatment experiments to determine the optimal therapeutic concentrations of sorafenib and nivolumab.By assessing cell viability and drug toxicity,we determined the half-maximal inhibitory concentration(IC50)of sorafenib for HepG2 liver cancer cell line as 11.16 μ M and for Huh7 liver cancer cell line as 10.70 μM,consistent with relevant literature reports.Furthermore,we observed that nivolumab at a concentration of 10 nM,in combination with human peripheral lymphocyte T cells,exhibited significant therapeutic effects on liver cancer cell lines,resulting in notable differences in tumor killing.Notably,we conducted experiments investigating the combined therapy of both drugs,which demonstrated that the combined application of sorafenib and nivolumab enhanced the killing efficacy against liver cancer cells,suggesting a synergistic effect of this drug combination and presenting a potential new strategy for liver cancer treatment.During the experiment,we encountered several issues.Firstly,the HepG2 cell line showed insensitivity to lenvatinib,prompting us to choose sorafenib as the targeted drug for subsequent experiments to align our research objectives with the therapeutic effects of the drug.Secondly,the Huh7 cell line exhibited poor viability during the printing process.To improve cell viability,we adjusted the formulation of the bioink to optimize the printing conditions.Additionally,we faced the challenge of extracellular leakage from the 3D bioprinted gel.To address this issue,we introduced sodium alginate-calcium chloride crosslinking agent to enhance gel stability and prevent cell leakage.This improvement ensured better cell fixation and protection during the printing process,thereby enhancing the reliability of the experimental results.We observed that nivolumab did not reach the IC50 for killing tumor cells,which could be attributed to the lack of specific T cells,affecting the drug’s effectiveness.To further investigate,we plan to utilize flow cytometry to screen for specific T cells and conduct additional experimental validation.Future research can apply our experimental findings to primary tumor specimens from patients,advancing the development of personalized medicine.By addressing the aforementioned issues,we will further enhance the application value of 3D bioprinting technology in liver cancer drug research and drive innovation in the field of liver cancer treatment.