The effect of prenatal exposure of a non-steroidal anti-inflammatory drug on the optic nerve of female rats: a stereological, histological, and electron microscopic study.

OBJECTIVE: Non-steroidal anti-inflammatory drugs (NSAIDs) can have adverse effects for in both mother and fetus following administration during the prenatal period. If given during pregnancy, diclofenac sodium (DS), an NSAID, is given during the pregnancy, may also affect the development of the central nervous system (CNS) or related structures. METHODS: Pregnant rats were separated into pure control (PG), saline (SG) and diclofenac groups (DG). A daily dose of 1 mg/kg of DS and 1 mL/kg saline was injected intraperitoneally to the DG and SG groups, respectively, from the 5th gestation day for a 15 day of period; the PG group received no treatment. After spontaneous delivery, female offspring were obtained from all groups. After the 20th week of postnatal life, the animals (n = 6 for each group) were perfused and the right optic nerves were resected. Sections were subjected to stereological and histological analysis. RESULTS: There were no significant differences (p > 0.05) between PG, SG and DG groups with respect to myelin thickness, axonal cross-sectional area, axon numerical density, total section area of optic nerve and axon number. CONCLUSIONS: Histological and stereological results indicated that treatment with DS or saline produced undesirable effects on female rat optic nerve development and myelinization with respect to morphology.

The protective effect of dehydroepiandrosterone on ovarian tissues after torsion-detorsion injury: a stereological and histopathological study.

OBJECTIVE: Ovarian torsion is a rare problem in pre-pubescent girls that must be included in the differential diagnosis of patients with abdominal or pelvic pain. Current advice for treatment to support ovary preservation considers the macroscopic appearance of the ovary, but this is not a reliable indicator of the degree of injury in cases of ovarian torsion. The aim of this study was to determine the effects of dehydroepiandrosterone (DHEA) treatment on the rat ovary after torsion-detorsion injury using a histopathological approach and stereological methods. MATERIALS AND METHODS: Fifteen adult female Sprague Dawley rats were divided into three groups: sham operated (Control; n=5), torsion-detorsion with saline (Saline; n = 5), and torsion-detorsion with DHEA (DHEA; n = 5). Rats in the sham-operated group underwent a surgical procedure similar to the other groups, but the adnexa were not torsioned. The DHEA group was injected intraperitoneally 3 hours before detorsion and saline was administered to the saline group. After 24 h of adnexal detorsion, the rats in all groups were sacrificed and the adnexa were removed. The volume of ovarian stroma and edema were estimated using the Cavalieri Principle and were applied to serial paraffin sections. Ovary sections were also evaluated histopathologically. RESULTS: The volume of ovarian edema was 35.4% lower in the saline group compared to the control group (p> 0.05). The volume of ovarian edema increased by 73.4% in the DHEA group (p<0.05). In the experimental groups, stromal volume was 41.0% higher in the saline group and 52% higher in the DHEA group in comparison to the control group. Statistically significant differences were found when comparing the volume of both ovarian edema and stroma between experimental groups (Detorsion/Saline group - Detorsion/DHEA group; p< 0.01). Histopathologically, mononuclear cell infiltration and vascular dilatation, perivascular edema and common necrotic changes were obvious in the torsion-detorsion damaged ovary. These changes were partially ameliorated by DHEA treatment. CONCLUSIONS: The present study shows that administration of DHEA has beneficial effects in the prevention of ischemia-reperfusion injuries of the ovaries.

Effect of high fat diet on the volume of liver and quantitative feature of Kupffer cells in the female rat: a stereological and ultrastructural study.

BACKGROUND: The role of Kupffer cells (KCs) in nonalcoholic steatohepatitis (NASH) which is regarded as a major cause of cryptogenic cirrhosis of the liver was investigated using stereological methods and electron microscopy in the rat model. To our knowledge, there is no stereological study on the volume of liver, total number, numerical density, and nuclear height of KCs of liver in the female rat fed with a high fat diet (HFD) in the literature. METHOD: 16 female Sprague Dawley rats were randomized into HFD and control group, with HFD and standard diet for 12 weeks, respectively. In this study, two basic research methods were used to analyze the samples. One was histopathological observation at both light and electron microscopic level. The other was stereological methods that consist of Cavalieri principle for liver volume estimation and physical disector method for estimation of numerical density and total number of KCs in the liver. RESULTS: Liver volume, both mean numerical density and total number of KCs, were statistically increased in HFD rats. Ultrastructurally, a significant decrease in the mean nuclear height of KCs in HFD rats was also found. In the control group, no abnormal change was observed, but in the HFD group, some changes such as diffuse steatosis, mononuclear cell infiltration, necrosis, fibrosis, accumulation of fat droplets and intra-cytoplasmic vacuoles, and swollen mitochondria with irregular membranes were observed in the hepatocytes. CONCLUSION: The number and activity of KCs are increased significantly in NASH induced by HFD, and KCs might be involved in the pathogenesis of steatohepatitis as previously attributed as a major cause of cryptogenic cirrhosis of the liver.

Effect of a high fat diet on quantitative features of adipocytes in the omentum: an experimental, stereological and ultrastructural study.

BACKGROUND: Omental adipose tissue specimens of female rats that were fed a high fat (HF) diet were evaluated stereologically and histopathologically. To our knowledge, there is no stereological study on numerical density, nuclear height and volume of adipocytes in omental adipose tissue in the female rat fed a HF diet in the literature. METHOD: 20 female Spraque Dawley rats were used in the study. 10 of the animals were fed HF diet consisting of 30% of calories from fat for 3 months. The remaining 10 rats, the control group, were fed a normal diet. After the experimental procedure, all animals were anesthetized and omental adipose tissues in the same area were dissected and fixed for the histochemical process using a mixture of 3% glutaraldehyde and 1% osmium tetraoxide in 0.1 M phosphate buffer. After embedding of tissues in araldite CY 212, semi-thin and thin sections were cut. The semi-thin sections were stained with toluidine blue. The physical dissector counting method was used for estimation of numerical density and nuclear height of adipocytes. Cavalieri principle was used for the estimation of adipocyte volume; volume fraction approach was applied to find the volume fraction of adipose tissue components. RESULTS: The mean numerical density of adipocytes in the HF diet group was significantly higher than the control. The mean nuclear height of adipocytes was also very high in the HF diet group. The volume fraction of adipose mass was increased whereas the extracellular matrix volume fraction was reduced in the HF diet group compared to the controls. The mean volume of adipocytes in the HF diet group was also significantly higher than in the control group. At the light microscopy level, it was found that adipocytes were enlarged and gaining irregular shape in the HF diet group. Thicker basal lamina and electron dense lipid content were also found in this group at the electron microscopy level. CONCLUSION: Lipid content and number of adipocytes in the adipose tissue of HF diet rats were higher than in the controls. Thus, HF diet induces increase in body weight via both hypertrophy and hyperplasia of adipocytes.