Dynamical changing patterns of histological structure and ultrastructure of liver graft undergoing warm ischemia injury from non-heart-beating donor in rats

Abstract

Aim: To investigate the histological and ultra-structural characteristics of liver graft during different of warm ischemia time (WIT) in rats and to predict the maximum limitation of liver graft to warm ischemia.

Methods: The rats were randomized into 7 groups undergoing warm ischemia injury for 0, 10, 15, 20, 30, 45 and 60 min, respectively. All specimens having undergone warm ischemia injury were investigated dynamically by light and electron microscopy, and histochemistry staining. After orthotopic liver transplantation (OLT), the recovery of morphology of liver grafts after 6, 24 and 48 h was observed.

Results: The donor liver from non-heart-beating donors (NHBD) underwent ischemia injury both in the warm ischemia period and in the reperfusion period. Morphological changes were positively related to warm ischemia injury in a time-dependent manner during the reperfusion period. The results demonstrated that different degrees of histiocyte degeneration were observed when WIT was within 30 min, and became more severe with the prolongation of WIT, no obvious hepatocyte necrosis was noted in any specimen. In the group undergoing warm ischemia injury for 45 min, small focal necrosis occurred in the central area of hepatic lobule first. In the group undergoing warm ischemia injury for 60 min, patchy or diffused necrosis was observed and the area was gradually extended, while hepatic sinusoid endothelial cells were obviously swollen. Hepatic sinusoid was obstructed and microcirculation was in disorder.

Conclusion: The rat liver graft undergoing warm ischemia injury is in the reversible stage when the WIT is within 30 min. The 45 min WIT may be a critical point of rat liver graft to endure warm ischemia injury. When the WIT is over 60 min, the damage is irreversible.

Figure 1

Cytoplasm loosening, cell edema, focal vacuole degeneration after reperfusion (10 × 10) in the group undergoing warm ischemia injury for < 30 min (A); obvious cell edema, ballooning-like degeneration after reperfusion (40 × 10) in the group undergoing warm ischemia injury for 30 min (B); focal necrosis around central lobule area after reperfusion (10 × 10) in the group undergoing warm ischemia injury for 45 min (C); plaque-like area necrosis after reperfusion (40 × 10) in the group undergoing warm ischemia injury for 60 min (D).

Figure 2

Swollen mitochondria, ruptured crista, rough endoplasmic reticulum degranulation (× 15 000) in the group undergoing warm ischemia injury for 30 min (A); phanero-swollen mitochondria and nuclear chromosome margination, and karyopyknosis after reperfusion (× 10 000) in the group undergoing warm ischemia injury for 45 min (B and C); significant swollen mitochondria, crista extinction, nuclear membrane rupture, karyolysis and karyorrhexis after reperfusion (× 5000) in the group undergoing warm ischemia injury for 60 min (D).

Figure 3

Cytoplasmic blebs and irregular endothelial sieve plate in some sinusoids after reperfusion (scanning electron microscope × 6000) in the group undergoing warm ischemia injury for 45 min (A); apoptosis and necrosis of hepatocytes after reperfusion (× 8000) in the group undergoing warm ischemia injury for 60 min (B); cytoplamic blebs, reticular cellulose and hemocytes after reperfusion (scanning electron microscope × 6000) in the group undergoing warm ischemia injury for 60 min (C); bleb or ballooning like swollen endothelial cells, blocked sinusoids and irreversible microcirculation disturbance after reperfusion (× 5000) in the group undergoing warm ischemia injury for 60 min (D).