Bone development in fetuses with intrauterine growth restriction caused by maternal endocrine-metabolic dysfunctions.

Intrauterine growth restriction (IUGR) affects a large proportion of infants, particularly in underdeveloped countries. Among the main causes of IUGR, maternal endocrine-metabolic dysfunction is highlighted, either due to its high incidence or due to the severity of the immediate and mediated changes that these dysfunctions cause in the fetus and the mother. Although the effects of endocrine and metabolic disorders have been widely researched, there are still no reviews that bring together and summarize the effects of these conditions on bone development in cases of IUGR. Therefore, the present literature review was conducted with the aim of discussing bone changes observed in fetuses with IUGR caused by maternal endocrine-metabolic dysfunction. The main endocrine dysfunctions that occur with IUGR include maternal hyperthyroidism, hypothyroidism, and hypoparathyroidism. Diabetes mellitus, hypertensive disorders, and obesity are the most important maternal metabolic dysfunctions that compromise fetal growth. The bone changes reported in the fetus are, for the most part, due to damage to cell proliferation and differentiation, as well as failures in the synthesis and mineralization of the extracellular matrix, which results in shortening and fragility of the bones. Some maternal dysfunctions, such as hyperthyroidism, have been widely studied, whereas conditions such as hypoparathyroidism and gestational hypertensive disorders require further study regarding the mechanisms underlying the development of bone changes. Similarly, there is a gap in the literature regarding changes related to intramembranous ossification, as most published articles only describe changes in endochondral bone formation associated with IUGR. Furthermore, there is a need for more research aimed at elucidating the late postnatal changes that occur in the skeletons of individuals affected by IUGR and their possible relationships with adult diseases, such as osteoarthritis and osteoporosis.

Three Cases of Mandibular Ossifying Fibroma in Cattle.

Neoplastic masses were evaluated in the rostral region of the mandible of three young adult cattle. In all three cases, the masses were macroscopically large, firm, ulcerated, infiltrative, whitish and solid, and led to tooth displacement and loss. Radiographically, the neoplastic masses were intraosseous and radiolucent with foci of radiopacity. Loss of radiopacity due to bone necrosis was seen in the mandibular bone adjacent to the neoplasms. Histologically, well-differentiated, infiltrative non-encapsulated mesenchymal neoplastic proliferation replaced the mandibular bone and extended to the oral mucosa in all three cases. The neoplastic cells had scant cytoplasm and fusiform or oval hyperchromatic nuclei with loose chromatin, and were arranged in bundles in various directions. Within the neoplastic tissue, there were mineralized bone trabeculae and unmineralized osteoid, lined by a layer of osteoblasts and osteocytes within the lacunae. The pre-existing bone tissue adjacent to the neoplasm had areas of necrosis and osteoclasis of variable extent and intensity. Based on the macroscopic, radiographic and microscopic findings, a diagnosis of mandibular ossifying fibroma was established in all three cattle.

Kisspeptin/Kiss1r system and angiogenic and immunological mediators at the maternal-fetal interface of domestic cats.

The Kisspeptin/Kiss1r system is a key regulator of reproduction by stimulating gonadotrophin-releasing hormone and luteinizing hormone release, and in vitro studies have shown that Kisspeptin can modulate angiogenesis and immune function, factors that are also essential for reproduction However, there are no studies on the expression of Kisspeptin/Kiss1r at the maternal-fetal interface in domestic cats and its relationship with angiogenic and immunological mediators. Thus, our objective was to evaluate the spatiotemporal expression profile of Kisspeptin/Kiss1r and angiogenic and immunological mediators in the uterus and placenta of domestic cats during pregnancy. Uterus and placenta samples were collected from cats in mid pregnancy (N = 6) and late pregnancy (N = 6), in addition to uterus from non-pregnant cats in diestrus (N = 7), to evaluate protein and gene expression of kisspeptin (Kiss1), kisspeptin receptor (Kiss1r), vascular endothelial growth factor (VEGF), tyrosine kinase receptor (Flk-1), placental growth factor (PLGF), interferon gamma (INFγ), migration inhibiting factor (MIF), tumor necrosis factor (TNFα), interleukins (IL6 and IL10) by immunohistochemistry and quantitative polymerase chain reaction. Pregnancy increased the uterine expression of Kiss1 and Kiss1r, especially at the late pregnancy, in addition to upregulating INFy, MIF, Vegf, Il10, and Tnf and downregulating Plgf. Higher placental expression of Kiss1r and Plgf mRNA occurred at the late pregnancy, while the expression of Kiss1, VEGF, Flk-1, INFy, TNFα, Il6, and IL10 was higher in the mid of pregnancy. A positive correlation between Kiss1 and Tnf was observed in the placenta, while Kiss1r had a negative correlation with Infγ, Il6, and Il10. The findings reveal that Kisspeptin/Kiss1r and angiogenic and immunological mediators at the maternal-fetal interface of pregnant cat have a gene correlation and are modulated by the gestational age. These data suggest possible functional links of Kisspeptin in placental angiogenesis and immunology.

Pyometra and estrous cycle modulate the uterine expression of the kisspeptin system and angiogenic and immune factors in cats.

Failures in hypothalamic kisspeptin/Kiss1r signaling are associated with infertility, and in vitro studies have shown that kisspeptin can modulate angiogenesis and immune activity. Because there is no in vivo research on the functional relationship between these factors in the reproductive system, especially in domestic cats, we evaluated the expression profile of kisspeptin/Kiss1r and angiogenic and immunological mediators in the genital tract of cyclic cats and of those with pyometra. The uterus of cats in diestrus exhibited greater gene and protein expression of Kiss1, as well as Vegf, Pigf, Mif, and Il6. In contrast, Kiss1r presented greater expression in proestrus/estrus, similarly to that observed for the immunostaining of INFγ, MIF, TNFα, and IL10. These factors were positively correlated with Kiss1 and/or Kiss1r, and a positive correlation between Kiss1 and Kiss1r was also observed in the uterus of cats during the estrous cycle. Cats with pyometra showed greater immunostaining of Kiss1 and Kiss1r on the endometrial surface and reduced immunostaining of Kiss1 in deep glands, whereas there was a significant reduction in Vegf, Pigf, Mif, and Il6 mRNA, and an increase in Tnf mRNA. The findings reveal that there is a gene correlation between kisspeptin/Kiss1r and angiogenic and immune mediators in the uterus of the domestic cat, which is modulated by the estrous cycle, and that pyometra affects the expression of these mediators. This study suggests, for the first time, a functional relationship between the Kiss/Kiss1r system and angiogenic and immune mediators in the female genital tract.

Nonlinear effects of caffeine on the viability, synthesis and gene expression of chondrocytes from the offspring of rats treated during pregnancy.

OBJECTIVE: Evaluate the effects of doses of caffeine administered to pregnant rats on the articular cartilage chondrocytes of their offspring. METHODS: Twenty-four adult Wistar rats were randomly assigned to four groups, with one control group and three groups being treated with caffeine at doses of 25, 50 and 100 mg/kg throughout pregnancy. At birth, three offspring/females were euthanized so that the chondrocytes could be extracted. At 7, 14 and 21 days of culture, the chondrocytes were subjected to the MTT cell viability assay and an evaluation of their alkaline phosphatase activity and collagen synthesis. Chondrocytes were also stained by Hematoxylin-eosin, PAS, Safranin-O and Alcian Blue. The Sox-9, Runx-2, aggrecan, collagen-II and alkaline phosphatase gene transcript levels were also evaluated. Mean comparisons were performed by the Student-Newman-Keuls test. RESULTS: Chondrocyte cultures from the 25 mg/kg group had the lowest results, as chondrocytes from this group had reduced viability, percentage of cells, alkaline phosphatase activity and collagen and chondrogenic matrix synthesis. A reduced expression of Sox-9, alkaline phosphatase and collagen-II was also detected in the 25 mg/kg group. Chondrocyte cultures of the group treated with 50 mg/kg caffeine showed reduced collagen synthesis and Sox-9 expression. The caffeine dose of 100 mg/kg also reduced collagen and Sox-9 and alkaline phosphatase expression. CONCLUSION: Caffeine administered to pregnant rats negatively alters the articular cartilage chondrocytes of their offspring, reducing the synthesis of collagen and Sox-9 expression regardless of the dose. This study also concluded that the effects of caffeine are not linear or dose-dependent.

Dose-dependent effect of triiodothyronine on the chondrogenic differentiation of mesenchymal stem cells from the bone marrow of female rats.

OBJECTIVES: Verify the in-vitro effect of triiodothyronine (T3) on the chondrogenic differentiation of female rat bone marrow mesenchymal stem cells (BMMSCs) over several time periods and at several doses. METHODS: CD54 + /CD73 + /CD90 +  BMMSCs from Wistar female rats were cultured in chondrogenic medium with or without T3 (0.01; 1; 100; 1000 nm). At seven, 14 and 21 days, the cell morphology, chondrogenic matrix formation and expression of Sox9 and collagen II were evaluated. KEY FINDINGS: The dose of 100 nm did not alter the parameters evaluated in any of the periods studied. However, the 0.01 nm T3 dose improved the chondrogenic potential by increasing the chondrogenic matrix formation and expression of Sox9 and collagen II in at least one of the evaluated periods; the 1 nm T3 dose also improved the chondrogenic potential by increasing the chondrogenic matrix formation and the expression of collagen II in at least one of the evaluated periods. The 1000 nm T3 dose improved the chondrogenic potential by increasing the chondrogenic matrix formation and Sox9 expression in at least one of the evaluated periods. CONCLUSIONS: T3 has a dose-dependent effect on the differentiation of BMMSCs from female rats.

Osteopetrosis in obese female rats is site-specifically inhibited by physical training.

NEW FINDINGS: What is the central question of this study? Clinical studies suggest that obesity 'protects' against osteoporosis. However, these studies used only bone densitometry and assessed only one bone site, which is insufficient to enable conclusions to be drawn about the response of the whole skeleton. Furthermore, the effects of exercise on bone responses in obesity have not been explored previously. What is the main finding and what is its importance? We show that obesity causes osteopetrosis. Therefore, the classical perspective of 'protective effects of obesity' needs to be reviewed, and exercise is an important tool to avoid these alterations and to maintain the homeostasis of bone. A sedentary lifestyle and obesity induce systemic inflammatory responses. Although the effects of physical inactivity on osseous tissue have been well established, the effects of obesity on bone tissue remain controversial. Furthermore, the effects of physical training on bone tissue responses in the presence of diet-induced obesity are unknown. Our aim was to investigate the effects of obesity and physical training at multiple bone sites in rats. Female Wistar rats were divided into the following four groups: (i) control diet, non-trained (C-NT); (ii) high-refined carbohydrate-containing diet, non-trained (HC-NT); (iii) control diet, trained (C-T); and (iv) high-refined carbohydrate-containing diet, trained (HC-T). At 5 months of age, the rats were submitted to daily exercise for 30 min day(-1). After 13 weeks, blood samples, adipose and skeletal tissues were harvested. Two-way ANOVA was applied to detect differences (significance accepted when P ≤ 0.05). The HC-NT group exhibited increased body mass, adiposity, serum leptin, serum insulin, insulin resistance index and concentrations of tumour necrosis factor-α and interleukin-6. Obese rats (HC-NT) exhibited thickening of nasal bones, trabecular bones in the lumbar vertebrae and long bones in a site-dependent manner. The HC-T group exhibited similar adiposity and inflammatory results. Morphological analysis of the lumbar vertebrae in rats fed the HC diet revealed characteristics of osteopetrosis that were inhibited by exercise. In conclusion, the HC diet induced obesity and inflammatory/hormonal alterations and increased the trabecular bone in a site-dependent manner. However, obesity caused osteopetrosis in the lumbar vertebrae, which could be inhibited by physical training. Although exercise inhibited the development of bone alterations, physical training did not inhibit the HC diet-induced obesity responses.

Effect of low-power gallium-aluminum-arsenium laser therapy (830 nm) in combination with bisphosphonate treatment on osteopenic bone structure: an experimental animal study.

Laser therapy is able to modulate cell metabolism and accelerate the repair of fracture. Little attention has been given to the effect of laser on bone with osteopenia or osteoporosis. The purpose of our study was to verify the effect of laser therapy in combination with bisphosphonate on osteopenic bone structure. The 35 Wistar female rats used were divided into five groups: (1) sham-operation rats (control), (2) ovariectomized (OVX'd) rats with osteopenia, (3) OVX'd rats with osteopenia treated with laser, (4) OVX'd rats with osteopenia treated with bisphosphonate and (5) OVX'd rats with osteopenia treated with bisphosphonate and laser. Groups 3 and 5 were given daily 6 mg doses of bisphosphonate orally. Groups 4 and 5 underwent low level laser therapy [gallium-aluminum-arsenium (GaAlAs) laser, 830 nm, 50 mW and 4 J/cm(2)] on the femoral neck and vertebral segments (T13-L2). Both treatments were performed over an 8-week period. Rats from the osteopenic control and osteopenic + laser groups presented marked osteopenia. In the osteopenic + bisphosphonate group, the trabecular bone volume in vertebra L2 was significantly greater than in the osteopenic control group. Notably, in the association between laser and bisphosphonate, the trabecular bone volume was significantly greater in vertebrae L2 and T13 and was similar to that in the sham-operation control group. It was concluded that the laser therapy associated with bisphosphonate treatment was the best method for reversing vertebral osteopenia caused by the ovariectomy.

Histomorphometry and expression of CDC-47 and caspase-3 in mammary glands of pregnant female rats with artificial hyperthyroidism.

The purpose of this study was to evaluate the effect of hyperthyroidism on mammary gland development and expression of two protein markers, CDC-47 for proliferation and caspase-3 for apoptosis in pregnant female rats. Thirty-six adult female Wistar rats were used in two groups: hyperthyroid and control. Rats were mated 60 days after the onset of thyroxine administration. Six animals/group were sacrificed on gestation days 7, 14, and 19. Artificial hyperthyroidism was induced by daily administration of thyroxine in the drinking water until the end of gestation. At the end of each period, rats were sacrificed, and their inguinal mammary glands were collected and processed for morphometric analysis. The percentages of epithelium, stroma, adipose tissue, and lacteal secretion were determined. Immunohistochemical analysis was also carried out using anti-CDC-47 and anti-caspase-3 antibodies to study proliferation and apoptosis, respectively. On the 19th day of gestation, thyroxine treatment significantly increased the percentage of mammary epithelium. Hyperthyroidism, however, did not change CDC-47 expression. The hyperthyroid group presented early lactogenesis and significantly larger lacteal secretion on the 19th day of gestation. There was no significant difference in caspase-3 expression between groups in any period. We may conclude that hyperthyroidism accelerates mammary gland development and increases lacteal secretion during gestation without increasing the proliferation rate and the expression of caspase-3.

Idiopathic hypertrophic osteopathy in a cat.

This report describes the first case of idiopathic hypertrophic osteopathy (HO) in a cat. No causes for the bone pathology were found following evaluation of the physical and laboratory examinations (complete blood count, albumin, creatinine, urea, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase and gamma-glutamyltransferase and urinalysis), and after histopathological evaluation of organs at necropsy. Based on the radiographic, clinical and anatomopathological findings, idiopathic HO was diagnosed.