Rosiglitazone, a thiazolidinedione peroxisome proliferator-activated receptor gamma ligand, reduces disease activity in nonalcoholic steatohepatitis (NASH), a disease associated with hepatocyte mitochondrial crystalline inclusions that are not seen in animal models of NASH. In human and animal studies of adipose tissue, thiazolidinediones may induce mitochondrial biogenesis and associated morphological changes. To determine if rosiglitazone alters the hepatocyte mitochondrial morphology in human NASH, we prospectively and systematically examined liver biopsies from human subjects with NASH before and after 48 weeks of rosiglitazone by transmission electron microscopy. Twenty patients (body mass index = 34 ± 7) were studied. Four coded sections from each of 20 pretherapy biopsies and each of 20 posttherapy biopsies were examined by transmission electron microscopy. The total hepatocyte mitochondria and crystal-containing mitochondria were counted, and semiquantitative scoring was performed for macrosteatosis, microsteatosis, dilated endoplasmic reticulum, apoptosis, Mallory bodies, and hepatocyte enlargement. The total mitochondria count was unchanged after therapy, but there was a significant increase in crystal-containing mitochondria from 4.0% (95% confidence interval = 1.8-8.8) to 7.2% (95% confidence interval = 3.9-12.6; odds ratio = 1.80; P = 0.04) after the treatment with rosiglitazone. Macrosteatosis (P < 0.001) and Mallory bodies (P = 0.05) significandy decreased, but no change was evident in microsteatosis, cellular enlargement, dilated endoplasmic reticulum, or apoptosis. Conclusion: Rosiglitazone therapy of NASH is associated with increased crystalline inclusions in hepatocyte mitochondria. Whether these are adaptive or pathological remains unknown, and further studies are warranted to assess hepatic mitochondrial function during thiazolidinedione therapy for NASH.