Management of carbon tetrachloride-induced acute liver injury in rats by syngeneic hepatocyte transplantation in spleen and peritoneal cavity

Abstract

Aim: Acute hepatitis may seldom have a fulminant course. In the treatment of this medical emergency, potential liver support measure must provide immediate and sufficient assistance to the hepatic function. The goal of our study was to study the adequacy of hepatocyte transplantation (HCTx) in two different anatomical sites, splenic parenchyma and peritoneal cavity, in a rat model of reversible acute hepatitis induced by carbon tetrachloride (CCl(4)).

Methods: After CCl(4) intoxication, 84 male Wistar rats used as recipients were divided in to four experimental groups accordingly to their treatment: Group A (n=24): intrasplenic transplantation of 10x10(6) isolated hepatocytes, Group B (n=24): intraperitoneal transplantation of 20x10(6) isolated hepatocytes attached on plastic microcarriers, Group C (n=18): intrasplenic injection of 1 mL normal saline (sham-operated controls), Group D (n=18): intraperitoneal injection of 2.5 mL normal saline (sham-operated controls). Survival, liver function tests (LFT) and histology were studied in all four groups, on d 2,5 and 10 post-HCTx.

Results: The ten-day survival (and mean survival) in the 4 groups was 72.2% (8.1+/-3.1), 33.3% (5.4+/-3.4), 0% (3.1+/-1.3) and 33.3% (5.4+/-3.6) in groups A, B, C, D, respectively (P(AB)<0.05, P(AC)<0.05, PBD=NS). In the final survivors, LFT (except alkaline phosphatase) and hepatic histology returned to normal, independently of their previous therapy. Viable hepatocytes were identified within splenic parenchyma (in group A on d 2) and both in the native liver and the fatty tissue of abdominal wall (in group B on d 5).

Conclusion: A significantly better survival of the intrasplenically transplanted animals has been demonstrated. Intraperitoneal hepatocytes failed to promptly engraft. A different timing between liver injury and intraperitoneal HCTx may give better results and merits further investigation.

Figure 1

Mean survival in all four groups (P_AC < 0.05, P_BD = NS, P_AB < 0.05).

Figure 2

Survival curves in all four groups. Notice that all animal deaths occurred within the first 6 d after poisoning.

Figure 3

Viable hepatocytes within splenic parenchyma, forming structures similar to liver plates on d 2 (the so-called “splenic hepatization”).

Figure 4

Microcarrier-cell aggregates within native liver parenchyma, Formation of neo-vessels in surrounding tissue on d 5.