Figure Legends
Figure 1 . Hemodynamics, vascular resistance and bile production of normothermic perfused cirrhotic and non-cirrhotic livers.(A) hepatic artery flow, (B) portal flow, (C) hepatic artery resistance, (D) portal resistance and (E) bile production of cirrhotic and non-cirrhotic livers measured during 360 min of normothermic perfusion. Data represent median and interquartile range in cirrhotic (n=7) and non-cirrhotic livers (n=4). Differences between groups were analyzed using the Mann-Whitney U test
Figure 2. Liver injury and liver function markers, measured in perfusate and bile during normothermic machine perfusion.(A) Perfusate AST and (B) ALT. Liver function parameters measured in perfusate, including; (C) lactate, (D) relation of perfusate lactate AUC and MELD score, (E) albumin and (F) total bilirubin. Liver function parameters measured in bile; (G) pH, (H) Δglucose perfusate vs bile, (I) Δbicarbonate bile vs perfusate and (J) bile bilirubin levels during 360 min of perfusion. Data represent median and interquartile range in cirrhotic (n=7). and non-cirrhotic livers (n=4). Differences in AUC between groups were analyzed using the Mann-Whitney U test; p value is presented in right corner of each graph
Figure 3. Histological characterization of liver biopsies taken pre- and post-perfusion. (A) Sirius red staining of a cirrhotic liver (50X) and (B) Sirius red staining of a non-cirrhotic liver (50X).(C) H&E staining of a -cirrhotic liver, before perfusion (200x), (D) non-cirrhotic liver pre-perfusion (200x), ( E) Cirrhotic liver after 6 hours of perfusion (200x) and (F) non-cirrhotic liver after 6 hours of perfusion (200x).
Figure 4. Schematic representation of the experimental set-up for studying pharmacokinetics of a cocktail of drugs and the effects of drug-drug interactions. Stable liver perfusion was maintained in the first 120 min of perfusion. Between 120-130 min, the drug cocktail was infused into the portal vein (1mL/min), during this time the hepatic extraction of the drugs was measured. Hepatic clearance as well as biliary clearance was measured for 120 min (120 -240 min). Thereafter, at t=240 min, a drug inhibitors were infused into the portal vein for 10 min (240 – 250). Between 245 – 255 the drug cocktail was infused into the portal vein to study drug-drug interactions. Perfusate and bile samples studying the extent of drug-drug interactions were taken between 245 and 365 min
Figure 5. Pharmacokinetic profiles of rosuvastatin, digoxin, metformin and furosemide in cirrhotic and non-cirrhotic perfused livers . Perfusate levels of (A) rosuvastatin (applied dose of 1.80 mg), (B) digoxin (applied dose of 0.11 mg), (C) metformin (applied dose of 74.40 mg), and (D) furosemide (applied dose of 0.77 mg) in cirrhotic and non-cirrhotic normothermic perfused livers. Biliary excretion profiles of the administered cocktail of drugs (E) rosuvastatin, (F) digoxin, (G) metformin and (H) furosemide. (I) hepatic extraction of drug cocktail compounds measured during dosing (n=3 non cirrhotic livers, n=4 cirrhotic livers). Data represent median and interquartile range in cirrhotic (n=7) and non-cirrhotic livers (n=4), unless indicated otherwise. Differences in AUC between groups were analyzed using the Mann-Whitney U test; p value is presented in right corner of each graph
Figure 6. Effect of drug inhibitor mix on hepatic clearance of rosuvastatin, digoxin, metformin and furosemide. (A) Graphical representation of relevant hepatic drug transporters for the victim drugs (rosuvastatin, digoxin, metformin and furosemide) and the applied perpetrators (quinidine, rifampicin, cimetidine, probenecid), (B) Ratio of perfusate Cmax and (C) AUCR with and without applied perpetrator drugs for the cirrhotic and non-cirrhotic livers. Data represent median and interquartile range in cirrhotic (n=7) and non-cirrhotic livers (n=4). Differences between groups were analyzed using the Mann-Whitney U test