Establishment of a 3D multicellular placental microtissues for investigating the effect of antidepressant vortioxetine.

The placenta is a unique organ with an active metabolism and dynamically changing physiology throughout pregnancy. It is difficult to elucidate the structure of cell-cell and cell-extracellular matrix interactions of the placenta in in vivo studies due to interspecies differences and ethical constraints. In this study, human umbilical cord vein cells (HUVEC) and human placental choriocarcinoma cells (BeWo) were co-cultured for the first time to form spheroids (microtissues) on a three-dimensional (3D) Petri Dish® mold and compared with a traditional two-dimensional (2D) system. Vortioxetine is an antidepressant with a lack of literature on its use in pregnancy in established cultures, the toxicity of vortioxetine was studied to investigate the response of spheroids representing placental tissue. Spheroids were characterised by morphology and exposed to vortioxetine. Cell viability and barrier integrity were then measured. Intercellular junctions and the localisation of serotonin transporter (SERT) proteins were demonstrated by immunofluorescence (IF) staining in BeWo cells. Human chorionic gonadotropin (beta-hCG) hormone levels were also measured. In the 3D system, cell viability and hormone production were higher than in the 2D system. It was observed that the barrier structure was impaired, the structure of intracellular skeletal elements was altered and SERT expression decreased depending on vortioxetine exposure. These results demonstrate that the multicellular microtissue placenta model can be used to obtain results that more closely resemble in vivo toxicity studies of various xenobiotics than other 2D and mono-culture spheroid models in the literature. It also describes the use of 3D models for soft tissues other than the placenta.

Developmental toxicity of benzyl benzoate in rats after maternal exposure throughout pregnancy.

The maternal and fetal toxicity of benzyl benzoate, commonly used as antiparasitic insecticide, was evaluated in pregnant rats after a daily oral dose of 25 and 100 mg/kg. Biochemical, histopathological, and morphological examinations were performed. Dams were observed for maternal body weights and food and water consumption and subjected to caesarean section on (GD) 20. Maternal and fetal liver, kidney, heart, brain, and placenta were examined histopathologically under light microscope. Maternal and fetal liver and placenta were stained immunohistochemically for vascular endothelial growth factor (VEGF). Morphometric analysis of fetal body lengths, placental measurements, and fetal skeletal stainings was performed. Statistically significant alterations in biochemical parameters and placental and skeletal measurements were determined in treatment groups. In addition to histopathological changes, considerable differences were observed in the immunolocalization of VEGF in treatment groups. These results demonstrated that benzyl benzoate and its metabolites can transport to the placenta and eventually enter the fetuses.

Does furan affect the thymus in growing male rats?

Furan has been identified in foods such as heat-treated foods, including coffee, canned meat, hazelnuts, and infant foods and formulas. Children may be exposed to furan via either consumption of these foods or their derivatives. We evaluated the effects of furan on the thymus of weaning male rats in the present study. Five separate groups containing male rats were used: control, oil control, and three furan-treated groups. Furan was given orally to rats in the treatment groups at doses of 2, 4, and 8 mg/kg/day for 90 days. At the end of the experiment, thymus of the rats were examined morphologically, histopathologically, and immunohistochemically. We observed that absolute and relative weights of thymus were decreased significantly in rats treated with 4- and 8-mg/kg/day doses of furan. In histopathological examination, enlargement of interstitial connective tissue between the thymic lobules, lymphocyte depletion, and hemorrhage were observed. We detected an increase in apoptotic cell counts in thymus of the treatment groups. In addition, we found significant differences in the distribution of fibronectin and transforming growth factor-beta in the thymus of the treatment groups. In conclusion, we suggest that furan has affected the thymus in growing male rats.

Toxicity of food contaminant furan on liver and kidney of growing male rats.

Furan is a chemical used in some industrial products and occurs naturally in heat-treated foods. We aimed to investigate the effects of orally administered furan on liver and kidney in growing Wistar male rats for 90 days. In this respect, biochemical, morphological, histopathological, and histomorphometrical examinations were performed. Three- to 4-week aged rats were divided into five groups of eight animals each; control, oil control; 2, 4, 8 mg/kg/day furan treatment groups. At the end of the experiment, antioxidant enzyme activities and serum AST, ALT, HDL, Urea, etc. levels were analyzed. Malondialdehyde (MDA) levels, superoxide dismutase (SOD), catalase (CAT), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) were also measured in liver homogenates. Also, liver and kidney were examined morphologically and histopathologically under light microscopy. According to the results of biochemical analysis, ALT, ALP, and LDL levels in treatment groups were significantly different compared with control groups. While LDL levels in treatment groups increased significantly, ALT and ALP levels decreased significantly. No significant changes were observed in liver MDA levels, superoxide dismutase and catalase activities in treatment groups. While IL-6 levels did not change in treatment groups, furan caused dose-dependent increases in liver TNF-α level of rats. In treatment groups, absolute and relative liver weights changed significantly, however, no significant changes were observed in kidney and relative kidney weights. Hyperemic blood vessels in the liver and congestion, edema, fibrosis, and tubular damage in the kidney of rats treated with furan were observed histopathologically. According to histomorphometric examinations, glomeruli diameters and glomerular volume decreased in the kidneys of rats in treatment groups.

Effect of patulin on the interdigitating dendritic cells (IDCs) of rat thymus.

Patulin is a common fungal contaminant of ripe apples used for the production of apple juice concentrates and it is also present in other fruits, vegetables and food products. Patulin is a secondary metabolite produced by species of the genera Penicillium, Aspergillus and Byssochlamys. Patulin has been reported to be mutagenic, carcinogenic and teratogenic. Antigen-presenting cells (APCs) are of prime importance in the innate immune response; they capture antigen in tissues and then migrate to the lymphoid organs to present the antigen to T lymphocytes. Thus, they are crucial for the initiation of immunity. Interdigitating dendritic cells (IDCs) are a subset of APCs that are present at the lymphatic organs. In the thymus, they act in positive and negative selection during T cell development. In the present study, patulin was administered orally to growing male rats aged 5-6 weeks. A dose of 0.1 mg kg(-1) bw day(-1) was given to rats for a period of 60 or 90 days daily. The effect of patulin on the IDCs of thymus was investigated by transmission electron microscopy (TEM), and the results were evaluated in terms of cell destruction. In the rats of the control group, it was observed that the IDCs had an indented nucleus, a clear cytoplasm and numerous membrane extensions. In the cytoplasm, a well-developed golgi complex, mitochondria, granular endoplasmic reticulum and a small number of lysosomal structures were observed. At day 60 of patulin-treated rat groups (P-60), loss of cristae in mitochondria and chromatin margination and lysis in the nucleus were found. It was observed that the IDCs had a perinuclear area of cytoplasm surrounded by a peripheral electron-lucent zone. In the cytoplasm of the 90-day patulin-treated rat group (P-90), a peripheral electron-lucent zone was also found, similar to the P-60 group. Additionally increase in vesicular and lysosomal structures, increase in apoptotic bodies and condensation of chromatin in the nucleus were noted. It was observed that patulin leads to apoptotic body formation and cell apoptosis in the IDCs of rat thymus especially in the P-90-treated groups.

Histological changes in the liver of fetuses of alcohol-treated pregnant rats.

In this study, immunolocalization of tenascin (TN), type IV collagen and histopathological changes in the liver of fetuses of pregnant Wistar albino rats treated with two doses of alcohol (1 and 5 g kg(-1) day(-1)) were determined. The samples of liver obtained from fetuses of rats on day 21 of gestation were evaluated morphologically and fixed for histology and immunohistochemistry. The sections stained with H&E and peroxidase-antiperoxidase (PAP) method for TN and type IV collagen were evaluated and immunohistochemical staining results were examined by the intensity of labelling. Statistical evaluation of data showed that there were no differences in liver weights between the control and calorie control groups; but some differences in relative liver weight between the control and alcohol treatment groups were determined. The changes in the liver histology such as degeneration of the cytoplasm of hepatocytes and increases in the number of megakaryocytes were seen in the fetuses of groups receiving alcohol during pregnancy. Increases in the immunolocalization of TN and type IV collagen in the liver of fetuses of alcohol-receiving rats were also found.

Effects of patulin on thymus capillary of rats.

Patulin is a mycotoxin that is produced by species of Penicillum, Aspergillus, and Byssochylamys molds that may grow on a variety of foods including fruit, grains and cheese. Patulin, at a dose of 0.1 mg kg(-1) bw day(-1) was administered orally to growing male rats aged 5-6 weeks for a period of 60 or 90 days. The dose of patulin used in the present study was based on estimated human exposure levels. At the end of these periods, the thymus glands of patulin-treated and control Wistar rats were removed and ultrastructural changes in capillary cells of the thymus of patulin-treated Wistar rats were determined by electron microscopy. The walls of thymus capillaries of the 60-day patulin-treated rat groups (P-60) exhibited degeneration observable in electron microscopic sections. For example, loss of cytoplasm and mitochondrial cristae of cells, swollen endothelial cells, increased thickness of the basement membrane, closed lumen of capillaries, accumulation of fibrous material at the periphery of the capillaries and nuclear anomalies were seen in these sections. Such degeneration and changes were also observed in sections of capillaries of the 90-day patulin-treated rat groups (P-90). The levels of degeneration of endothelial cell nucleus of P-90 were greater than those of P-60. This study demonstrated the ultrastructural degeneration of thymus capillary cells of patulin-treated rats. The results obtained from this study may provide a guide to research dealing with the toxic effects of patulin on tissue and organ ultrastructure.

Subacute toxicity of celecoxib on thyroid and testis of rats: Hormonal and histopathological changes.

Celecoxib is an effective agent in the treatment of signs and symptoms of inflammation, rheumatoid arthritis and osteoarthritis. The purpose of this study is to assess the effects of two different doses of celecoxib on some hormones and endocrine glands of male rats. In this study, the doses of 10 and 50mg/kg/day of celecoxib were given to male rats orally for 28 days. At the end of the study, serum total triiodothyronine (T(3)), total thyroxine (T(4)), thyroid stimulating hormone (TSH), testosterone and luteinizing hormone (LH) levels of rats were analyzed by radioimmunoassay technique using RIA kits. Thyroid and testis tissues of male rats were examined histopathologically. While there was no a change in serum T(3), T(4) and LH levels of celecoxib-treated rats, there were differences in serum TSH and testosterone levels of rats treated with 50mg/kg/day celecoxib for 28 days compared with those of control rats. In histopathological examinations, celecoxib-related changes were found in thyroid glands of the rats.

Polidocanol at different concentrations for pleurodesis in rats.

PURPOSE: We previously found that 0.5% polidocanol was more effective than tetracycline for pleurodesis in rats. Thus, we conducted the present study to evaluate the efficacy of different concentrations of polidocanol for pleurodesis in rats. METHODS: We divided 54 albino Wistar rats into six groups. Groups 1, 2, and 3 were given isotonic saline, 35 mg/kg tetracycline, and 0.6 mg of diluted polidocanol, respectively, being the daily recommended dose for humans. Groups 4, 5, and 6 were given 0.5%, 1%, and 2% polidocanol, respectively. All solutions were given intrapleurally in a volume of 0.5 ml. We examined the rats for macroscopic pleural adhesions and compared the mean values of macroscopic scoring among the six groups. RESULTS: The rats given polidocanol and tetracycline had significantly more adhesions than the control group, and polidocanol at concentrations of 0.5%, 1%, and 2% was more effective for pleurodesis than tetracycline. The diluted polidocanol was not more effective than tetracycline. There was no difference between the effects of the 0.5% and 1% concentrations, but the 2% polidocanol group had significantly more adhesions than the other groups. CONCLUSIONS: Polidocanol at concentrations of 0.5%, 1%, and 2% was a more effective sclerosing agent than tetracycline for pleurodesis. While 2% polidocanol was the most efficient sclerosing agent, the daily maximum recommended dose of polidocanol for humans was not more effective than tetracycline.

The role of fibrin tissue adhesives in flap necrosis in rats.

Flap necrosis is an important issue in surgery, and fibrin tissue adhesives, due to beneficial properties in preventing flap necrosis, were used in this study. Two groups, each comprising of 10 rats, were formed. Group I served as a control group, and fibrin tissue adhesive was applied to group II. The fibrinogen and thrombin concentrations in fibrin tissue adhesive were 30 mg/ml and 10 U/ml, respectively. The mean area of flap necrosis was 687.5 +/- 72.5 mm2 and 78.5 +/- 11.0 mm2 in the control and fibrin tissue adhesive groups (p < .0001), respectively. The percentage of flap necrosis was significantly lower in the fibrin tissue adhesive group compared to the control group (5.6% vs 49.1%) (p < .0001). Fibrin tissue adhesives decreased flap necrosis significantly compared to the control group.

Dose-dependent effects of carbendazim on rat thymus.

In this study, the effects of carbendazim on the thymus in male rats were evaluated. Carbendazim was administered at 0, 150, 300 and 600 mg kg(-1) day(-1) doses by gavage to male rats for 15 weeks. Body weights of rats in all groups were recorded weekly during treatment. At the end of the experiment, the effects of carbendazim on the thymus were investigated histopathologically and morphologically. Also, based on these effects, change in immunolocalization of fibronectin (FN), which is a component of the extracellular matrix, was investigated immunohistochemically. Fibrosis and oedema were observed in the thymus of rats treated with 300 and 600 mg kg(-1) day(-1) doses of carbendazim. Also in this region, an increase in FN density was noted at the end of the immunohistochemical investigation. A decrease was observed in absolute and relative thymus weights of rats treated with carbendazim compared with the control group. While the decrease in absolute thymus weight was statistically significant in rats exposed to carbendazim at the highest dose, the decrease in relative thymus weights was statistically significant for all carbendazim doses.

Investigation of the effects of patulin on thyroid and testis, and hormone levels in growing male rats.

Patulin is a mycotoxin produced by several species of Penicillium, Aspergillus and Byssachlamys. Patulin can be produced on different food products including fruits, grains, cheese, cured meats, but in natural situations patulin is usually found in apple and apple products. In the present study, the time-dependent effects of patulin on the T3, T4, thyroid stimulating hormone, testosterone, luteinizing hormone and growth hormone levels of growing male rats were investigated. Patulin, at a dose of 0.1 mg/kg bw/day, was administered by gavage to growing male rats aged 5-6 weeks for a period of 60 or 90 days. The dose of patulin used in the present study was based on estimated human exposure levels. At the end of the experiment, serum T3, T4, TSH, testosterone, LH and GH levels of rats in control and treatment groups were analysed. In addition, the thyroid and testes were histopathologically examined by light microscopy. Results revealed that while patulin caused an increase (66.6%) in testosterone levels and a decrease (17.3%) in T4 levels of rats treated for 60 days, there was no change in the other hormone levels compared to those of the control group. When patulin treatment was extended to 90 days, increased serum testosterone (75%) and LH levels (146%) were observed. In histological examinations of the testes of rats treated with patulin, oedema, fibrosis and local Leydig cell hyperplasia in the interstitial tissue, and disorganization of seminiferous tubule epithelium were also observed. In addition, the thyroid of rats treated with patulin revealed lymphoid cell inflitration and enlargement of interstitial tissue between follicles, and degenerated colloid.

The efficacy of polidocanol in pleurodesis in rats.

PURPOSE: Pleurodesis is used to treat pleural effusions, and a number of agents with varying degrees of efficacy and systemic toxicity have been trialed. This study was conducted to evaluate the efficacy and systemic toxicity of polidocanol in pleurodesis. METHODS: Thirty albino Wistar rats were divided into three groups of ten rats each. Group 1 (control) was given isotonic saline, group 2 was given 35 mg/kg tetracycline, and group 3 was given 2.5 mg 0.5% polidocanol, all intrapleurally in a total volume of 0.5 ml. The rats were killed on postoperative day 30 and the macroscopic pleural adhesions and microscopic evidence of inflammation were evaluated. Hepatic, renal, and pancreatic function tests were done and various tissues were microscopically examined to detect systemic toxicity. The mean values of macroscopic and microscopic scoring and biochemical parameters were compared among the three groups. RESULTS: The polidocanol- and tetracycline-treated rats had significantly more adhesions than the control group rats, and polidocanol was more effective for pleurodesis than tetracycline (P = 0.027). Microscopic scoring was similar in the polidocanol- and tetracycline-treated rats, being significantly higher than that in the control rats. No significant difference was found in the biochemical parameters among the three groups. There were no signs of toxicity in any of the tissues studied microscopically. CONCLUSIONS: Polidocanol was found to be a more effective sclerosing agent than tetracycline for pleurodesis. Systemic toxicity was not shown by the biochemical parameters and histopathologic findings.

Efficacy of topical nitroglycerin and transcutaneous electrical nerve stimulation on survival of random-pattern skin flaps in rats.

We compared the efficacy of topical nitroglycerin and transcutaneous electrical nerve stimulation (TENS) on the survival of random-pattern skin flaps in rats. Thirty Wistar albino rats were used and a dorsal, cranially-based random-pattern flap was raised. The rats were divided into three groups of 10 rats each. The first group had only the flap raised while the second and third groups were given topical nitroglycerin 5 mg or TENS for one hour a day for seven days. The amount of flap necrosis was measured on the seventh postoperative day. The mean area of necrosis in the flaps were 726.2, 544.2, and 150.0 mm2 in the control, nitroglycerin, and TENS groups, respectively. The mean percentage of flaps that necrosed were 51.9, 38.9, and 10.7 in the corresponding groups. The TENS group had significantly higher percentage area of flap surviving than the control (p < 0.0001) and nitroglycerin groups (p = 0.002). TENS, with its efficacy on survival and with negligible side-effects, could be a reliable treatment. Clinically, it can easily be used postoperatively when flaps become ischaemic, and will be tolerated well by patients.