Morphometric Study
The morphometric study was conducted by two independent
histopathologists experienced in animal models. They were not aware of
which animal group the images come from and analyzed each criterion
separately. Lesions identified in each organ were described according to
the International Harmonization of Nomenclature and Diagnostic Criteria
(INHAND) (16, 17).
A semiquantitative scoring analyses of electron microscope images was
used to evaluate the degree of ultrastructural damage, as previously
described by Sergazy et al., 2020, which utilizes a four-point scale
(Grade 1–4) for the morphological assessment of subcellular structures
(18, 19). In brief, slides assessed for different morphologic changes,
and lesions were classified as present/absent on a scale from 0 to 1, or
assessed for severity on a scale from 0 to 4. Histopathological lesions
of the heart evaluated as present – ”1” or absent – ”0”, included
cardiomyocyte hypertrophy, focal hyper-eosinophilia, enlargement of
cardiomyocyte nuclei/multinucleated cardiomyocytes, cellular infiltrate,
and atherosclerosis. Fibrosis (perivascular and interstitial) was
assessed on a scale: ”0” – none, ”1” – minimal changes, ”2” – mild
changes, ”3” – moderate changes, ”4” – severe lesions. Kidney lesions
evaluated as present – ”1” or absent – ”0”, included tubular
epitheliocyte vacuolization and hyperplasia, formation of protein
cylinders in tubules, interstitial cellular infiltrate, thickening and
fibrosis of glomerular membranes, hyaline glomerulopathy, and
glomerulosclerosis. Liver lesions evaluated as present – ”1” or absent
– ”0”, included focal hepatocyte vacuolization, presence of
hyperchromatic/double-nucleated hepatocytes, lymphocytic infiltration,
hyperplasia (cysts) of bile ducts, and periportal fibrosis. Glycogen
accumulation was differentiated on a scale from 0 to 3: ”0” – less than
10% of hepatocytes, ”1” – 11-30% of hepatocytes, ”2” – 31-60%, ”3”
– more than 61% of hepatocytes.