Background & Aims
Hepatocellular ballooning is a key finding in nonalcoholic steatohepatitis (NASH).
It is conventionally defined by hemotoxylin and eosin (H&E) staining showing enlarged
cells with rarefied cytoplasm and recently by changes in the cytoskeleton. Fat droplets
are emerging as important organelles in cell metabolism. To address a possible relation
between fat droplets and ballooning, we studied fat staining, H&E, and keratin 18
staining in human NASH.
Methods
Sequential staining and high resolution imaging were used to study freshly prepared
cryo-sections from 10 patients with histologically confirmed steatohepatitis using
oil red O for fat droplet identification, H&E to identify ballooning, and anti-K18
to confirm cytoskeletal changes. High resolution images were captured at each stage
using the Aperio Scanscope. To provide ultrastructural correlation, glutaraldehyde-fixed
specimens were studied using transmission electron microscopy (TEM) with serial sectioning
for localization of ballooned cells by light microscopy and TEM in identical specimens.
Results
Serial staining consistently demonstrated that hepatocellular ballooning is associated
with fat droplet accumulation evident by oil red O positivity and depletion of cytoplasmic
keratin 18 with K-18 positive Mallory–Denk bodies (MDB). TEM confirmed the association
between osmium stained fat droplets, MDB formation, and cellular enlargement and suggested
droplet-associated dilation of the endoplasmic reticulum.
Conclusions
These results indicate a relationship between cellular ballooning, fat droplet accumulation,
and cytoskeletal injury in NASH. We speculate that injury to multiple, organelles
including fat droplets and endoplasmic reticulum, contribute to this characteristic
finding.
Keywords
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Article info
Publication history
Published online: July 01, 2010
Accepted:
April 15,
2010
Received in revised form:
March 25,
2010
Received:
February 1,
2010
Identification
Copyright
© 2010 European Association for the Study of the Liver. Published by Elsevier Inc. All rights reserved.
