Construction Of Bionic Bioartificial Liver And Its Effect On Acute Liver Failure

Purpose:Based on the hepatocyte microcarriers and semi permeable membrane microtubules,the bionic liver lobule structure unit was constructed;based on the liver lobule structure and function unit,the liver chip was constructed;the application of liver-chip in the treatment of acute liver failure with bionic bioartificial liver system(BBALS)was studied.Methods:The first part is to construct a liver microarray based on the structural and functional units of hepatic lobules.By evaluating the liver function of hepatocyte microcarriers,the possibility of simulating the liver microenvironment and acting as an in vitro bioreactor unit was verified,and the hepatocyte microcarriers were combined with semi permeable membrane microtubules to construct the structural and functional units of hepatic lobules.By optimizing the different flow rates of the two-way circulation system,the most suitable flow rate for hepatocyte microcarrier culture was selected,which can maintain the strongest activity and the best cell function of hepatocyte microcarrier.The experiment was divided into two groups:two-dimensional plate culture group and three-dimensional culture group.The glycogen synthesis was observed by periodic acid Schiff stain(PAS),and the uptake and excretion capacity were observed by indocyanine green(ICG)staining,The expression of cytochrome P-450(cyp-450)was detected.The optimal flow rate was determined by cell proliferation and liver function.The second part:through the use of the constructed bionic bioartificial liver,the effect of its treatment of acute liver failure was studied.3D liver microarray was used as a new bioreactor in bbals.The rabbit model of acute liver failure(ALF)was established by filtering D-galactosamine(D-gal),and plasma exchange was performed to study the therapeutic effect of bbals on ALF.The levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),urea and albumin were observed.The degree of inflammation,proliferation and apoptosis were detected by H&E,Ki-67 and TUNEL staining.Results:Part one:compared with the plate culture group,the microcarrier 3D culture group had more glycogen synthesis,faster ICG uptake and release,higher urea synthesis,higher albumin secretion and higher cyp-450 levels.The optimal flow rate was obtained,which proved that liver chip could maintain high cell activity and specific function at 5mL/h.Part II:compared with untreated ALF group,ALT and AST were significantly decreased,urea and plasma production were significantly improved,inflammatory infiltration,hepatocyte apoptosis and hepatocyte proliferation were significantly reduced in bbals treated group.Conclusions:1.Compared with cell plate culture,hepatocytes cultured on microcarrier 3D have better liver reserve,synthesis,secretion and metabolism ability.The functional unit simulating liver lobule structure was constructed by assembling hepatocytes with semi permeable membrane microtubules.It was found that the liver microarray constructed by the structure and function units of hepatic lobule combined with bidirectional circulation system could achieve the best growth activity at the flow rate of 0.5mL/h,and had good liver specific function.2.After the treatment of acute liver failure with bionic bioartificial liver,the degree of liver damage is reduced,and the detoxification and synthesis functions are restored to a certain extent,which shows its application prospect in tissue engineering.