Effect of cold aerobic perfusion on nonparenchymal cell viability of rat livers.

We investigated the effect of oxygen supply on hepatic cellular viability during cold perfusion storage of rat livers. A perfluoro-N-methyldecahydroisoquinoline (FMIQ) emulsion is used as an oxygen carrier. The composition of the perfusate containing 20 w/v% FMIQ is essentially the same as the University of Wisconsin (UW) solution except for the exclusion of hydroxyethyl starch. Rat livers were perfused at 4 degrees C for up to 24 h with either UW solution (group I, oxygenated; group II, unoxygenated) or FMIQ solution (group III, oxygenated; group IV, unoxygenated). After perfusion storage, the livers were reperfused with warm (37 degrees C) oxygenated or cold (4 degrees C) unoxygenated Krebs-Henseleit bicarbonate buffer, and nuclear trypan blue uptake was measured as the index of cell death. With warm oxygenated reperfusion, there remained less than 2% noviable parenchymal cells up to 24 h, regardless of perfusate or oxygenation. In UW-perfused livers, the proportion of nonviable nonprenchymal cells (NPC) increased progressively regardless of oxygenation, the values in groups I and II in the periportal field at 24 h being 39.9 +/- 4.7% (mean +/- SD) and 36.5 +/- 4.2%, respectively. By contrast, in FMIQ-perfused livers, dye uptake by NPC was significantly reduced with oxygenation (16.9 +/- 5.7% and 39.4% +/- 9.1% at 24 h in groups III and IV; P < 0.001). With cold unoxygenated reperfusion, livers in groups I, II, and IV showed a significant decrease of nonviable NPC, while those in group III showed no significant changes. These data indicate that oxygen supply during perfusion storage of the liver may ameliorate lethal injury to NPC precipitated during reperfusion.

[The effect of oxygen supply in continuous cold perfusion of the rat liver using perfluorochemical emulsion--parenchymal and nonparenchymal cell injuries evaluated with trypan blue perfusion/fixation techniques].

We investigated the effect of hypothermic aerobic perfusion of the rat liver using perfluoro-N-methyl-decahydroisoquinoline (FMIQ) on the cell viabilities of hepatic parenchyma and nonparenchyma in comparison with University of Wisconsin (UW) solution. Rat liver were perfused at 4 degrees C up to 24hr at a flow rate of 0.2 ml/g liver tissue/min with either UW or a perfusate (m-FMIQ) containing 20% FMIQ with or without oxygen saturation. After storage, livers were perfused with Krebs-Henseleit buffer containing trypan blue (200 microM). Nuclear dye uptake was measured as an indicator of cell death. Dye uptake of parenchymal cells remained less than 2% for up to 24hr, regardless of perfusate or O2-saturation. In UW perfused livers, nonparenchymal cells showed time related increases of dye positive nuclei, which were not affected by O2-saturation (39.9 +/- 4.7% and 36.5 +/- 4.2% at 24hr in O2-saturated and nonsaturated groups, respectively). In contrast, we found excellent protective effects of oxygen supply with m-EMIQ on the nonparenchymal cell viability, the dye uptake values being 16.9 +/- 5.7% and 39.4 +/- 9.1% at 24hr in O2-saturated and nonsaturated groups, respectively (p < 0.001). In conclusion, oxygen supply during hypothermic perfusion may be useful in preventing nonparenchymal cell injury of the liver, thereby leading to dramatic improvement of the hepatic microcirculation.

Intraarterial infusion of high-dose adriamycin for unresectable hepatocellular carcinoma using direct hemoperfusion under hepatic venous isolation.

A 67-year-old man with an extensive hepatocellular carcinoma (HCC) was treated successfully with intraarterial infusion of high-dose adriamycin (ADR), 150 mg/m2, five minutes continuous infusion using an extra-corporeal system consisting of direct hemoperfusion (DHP) under hepatic venous isolation (HVI). During drug infusion, hepatic effluent was isolated and adsorbed by the DHP for 30 mins. Plasma ADR levels in the radial artery reached a peak of 2.00 micrograms/ml at five mins after the initiation of drug infusion. Peak values at the inlet and outlet of the DHP were 19.71 micrograms/ml and 1.75 micrograms/ml, respectively, indicating substantial drug adsorption by the DHP. The estimated drug removal rate was 31.1%. This treatment led to a marked regression of tumors with tolerable systemic toxicities. Although the patient subsequently died 9 months after treatment of progression of disease, this treatment resulted in a remission of significant duration.

[A comparative study of tumor responses of hepatocellular carcinoma and colorectal hepatic metastasis to cytotoxic anticancer drug].

We investigated the characteristics of tumor responses of hepatocellular carcinoma (HCC, n = 14) and colorectal hepatic metastasis (CHM, n = 10) to high dose hepatic arterial chemotherapy using direct hemoperfusion under hepatic venous isolation. Adriamycin was given at doses ranging from 100 to 150 mg/m2. During drug infusion, two patients with HCC and three with CHM were treated by balloon-occluded arterial infusion (BOAI). Computerized tomographic findings after treatment were compared between the two diseases. In addition, response characteristics in each disease were investigated in relation to tumor factors such as size, capsule formation, vascularity and macroscopic type. Eight (57%) out of 14 patients with HCC showed either size reduction (n = 5) or liquefaction (n = 3), while the remaining six patients showed no apparent changes. On the other hand, seven (70%) of 10 patients with CHM showed liquefaction without any evidence of tumor size reduction. Regardless of the disease, all patients treated with BOAI had liquefaction as a sign of tumor responses. Based on these observations, it is concluded that response characteristics may differ depending on tumor factors such as vascularity and the nature of the treatment as represented by BOAI.

[Comparison of hepatic tissue extraction rates of cytotoxic anticancer drugs during hepatic arterial chemotherapy--evaluation using direct hemoperfusion under hepatic venous isolation].

Hepatic extraction rates (ER) of anticancer drugs during hepatic arterial infusion were investigated with the aid of direct hemoperfusion (DHP) under hepatic venous isolation (HVI). Using mongrel dogs (n = 6), adriamycin (ADR), mitomycin C (MMC) and cisplatin (CDDP) were simultaneously administered to the hepatic artery at each dosage of 1 mg/kg in 10 minutes under HVI.DHP. Hepatic venous flow and plasma concentrations of each drug at the carotid artery, the inlet and outlet sides of DHP were periodically determined during HVI.DHP. Based on these data, drug adsorption and removal rates were estimated. In addition, hepatic tissue uptakes of each drug were calculated from the amounts of drug administered and leaked in the hepatic effluent. Subsequently, the percentage of tissue uptake of each drug to the amount of drug administered was determined as ER of each drug. Drug adsorption rates during the first 10 minutes after infusion showed no significant difference among three drugs. Drug removal rate of CDDP tended to be higher than those of other two drugs. ER of CDDP (54.8 +/- 18.3%) were significantly lower (p < 0.01) as compared to ADR (84.4 +/- 16.2%) and MMC (83.1 +/- 15.7%). These results indicate that ER of each drug should be taken into consideration to determine appropriate drug for hepatic arterial chemotherapy.

Suprahepatic vena cava anastomosis of the donor liver to the recipient retrohepatic vena cava in canine liver transplantation.

A revised method of the upper caval anastomosis in canine orthotopic liver transplantation is described. It uses the recipient retrohepatic inferior vena cava below the suprahepatic veins for vascular suturing. Ten consecutive operations were performed to assess the feasibility of this method, with special reference to the outflow obstruction at the level of the suprahepatic inferior vena cava. Seven of 10 dogs survived more than 6 days. The cause of death was not related to the outflow obstruction in any instance. Regardless of the duration of survival, free hepatic vein pressures as well as portal vein pressures of all dogs remained in the physiological range even after the skin closures. Based on these observations, we conclude that this approach is safe and reproducible in experimental transplantation of the canine liver.

[A new method of intraarterial high dose chemotherapy for unresectable hepatomas using direct hemoperfusion under hepatic venous isolation (HVI.DVP)].

We have recently developed a simple method of high dose chemotherapy using HVI.DHP. The effect of HVI.DHP was evaluated in the elimination of anticancer drugs. Adriamycin (ADR: 3mg/kg), mitomycin C (MMC:1mg/kg) and cisplatin (CDDP: 2mg/kg) were given to mongrel dogs through the hepatic artery with or without HVI.DHP. The drug removal rates were similar in three drugs. Area under the curve and peak systemic level of each drug were substantially reduced by means of HVI.DHP, the values of ADR, MMC and CDDP being 17.4 and 6.2%, 4.5 and 5.9%, and 24.6 and 23.1%, respectively, of the corresponding values of simple hepatic artery infusions. Ten patients with unresectable hepatomas were treated by intraarterial high dose ADR (100-150mg/m2) using HVI.DHP. All except for two patients with catheter related complications recovered uneventfully and showed marked reductions of serum alpha-fetoprotein. Complete response was observed in one and partial response in five patients, for an overall response rate of 60%. Systemic toxicities of ADR were minimized: no cardiotoxicity, leucopenia less than 1000 cells/mm3 in one and mild hair loss in six patients. On the basis of these results, we consider this method to be an attractive therapeutic option for hepatic tumors.