Nandrolone decanoate in induced fracture nonunion with vascular deficit in rat model: morphological aspects.

BACKGROUND: The nonunion fracture is a relatively frequent complication in both human and veterinary medicine. Specifically, atrophic fracture nonunions are difficult to treat, with revision surgery usually providing the best prognosis. Anabolic steroids, such as nandrolone decanoate (ND), have been reported to have beneficial clinical effects on bone mass gain during osteoporosis; however, their utility in promoting regeneration in atrophic nonunions has not been documented. Our objective was to examine morphological changes induced by the ND in experimental fracture nonunion with vascular deficit in the rat model. METHODS: Fourteen adult Wistar rats had an atrophic fracture nonunion induced in the diaphysis of their left femur. Rats were allocated into two groups: control group and nandrolone decanoate group. Rats in the latter group were given nandrolone decanoate (1.5 mg/kg IM, once a week, during 4 weeks after confirmation of fracture nonunion radiographically). Radiographic and anatomopathological examination, micro-tomography and histological analysis were assessed to characterize the morphological changes promoted by the nandrolone decanoate use. RESULTS: Based on radiology, anatomopathological evaluation, computed micro-tomography and conventional microscopy, nandrolone decanoate promoted bone regeneration at the fracture nonunion site by increasing the cellularity at the fracture site. Percentage of collagen was not significantly different between groups, consistent with high-quality regenerated bone. CONCLUSION: The anabolic steroid nandrolone decanoate improved bone mass and regeneration without affecting collagen production and therefore has potential for improving outcomes for atrophic fracture nonunion.

Combination therapy of canine osteosarcoma with canine bone marrow stem cells, bone morphogenetic protein and carboplatin in an in vivo model.

Osteosarcoma (OSA) is the most common malignant bone cancer in children and dogs. The therapeutic protocols adopted for dogs and humans are very similar, involving surgical options such as amputation. Besides surgical options, radiotherapy and chemotherapy also are adopted. However, hematologic, gastrointestinal and renal toxicity may occur because of chemotherapy treatments. Recent study clearly showed that mesenchymal stem cells (MSCs) combined with recombinant human bone morphogenetic protein (rhBMP-2) may be associated with decreases of the tumorigenic potential of canine OSA. The aim of this study was to analyse the efficacy of chemotherapy with carboplatin and rhBMP-2 with MSCs in a canine OSA in vivo model. Canine OSA cells were implanted in mice Balb-c/nude with MSCs, rhBMP-2 and carboplatin. Flow cytometry and PCR for markers involved in tumour suppression pathways were analysed. Results showed that the combination of MSCs and rhBMP-2 reduced tumour mass and infiltration of neoplastic cells in tissues more efficiently than carboplatin alone. Thus it was demonstrated that the use of rhBMP-2 and MSCs, in combination with conventional antineoplastic, may be an efficient treatment strategy.

Morphological study of the eye bulb of the hammerhead shark, Sphyrna lewini (Elasmobranch: Carcharhinidae) / Estudio morfológico del bulbo ocular del tiburón martillo, Sphyrna lewini (Elasmobranch: Carcharhinidae)

The hammerhead sharks shows a head laterally expanded with eyes and nostrils on the edges, which gives the species a hammer appearance. Another strand of studies indicates that the hypothesis of head shape alterations for better adaptions in the sensorial system with the development of structures associated to binocular sight and others sensorial organs. Given the variety of characteristics, the study aim was to describe the anatomical constitution of the hammerhead shark eye bulb. The bulb and its annexes exenteration was performed, and further dissection; the morphological description of the muscle insertions and eye bulb components were based on direct observation and were further photographed and catalogued. The eye bulb fragments and its annexes were histological technical. Between the sclera and the choroid it was possible to observe, by the electronical scanning microscopy, the thin layer called supra-choroid, in this region, spaces often filled by lymphatic vessels allied to a matrix formed by loose conjunctive tissue are found. In the choroid, a layer which is rich in blood vessels, loose conjunctive tissue and collagen fibers, was observed, besides pigmentary cells full of melanin in its interior, which result in its layer's dark color. Ciliary body is a choroid's dilatation; it has the aspect of a thick ring in finger-like shape, pigmented, covering the sclera surface and containing pigmentary cells. The crystalline capsule, which shows an acellular covering that, is hyaline and homogeneous. In the electronical scanning microscopy, it was observed that the capsule is extremely thick especially in the anterior face. The capsule is very elastic, constituted mainly by thin lamellae of collagenous fibers, as illustrated by the electronically scanning microscopy. Anatomic variations related mainly to the position of the eye bulb in the skull, fibrous tunica and lens call the attention and must be related to its habitat.

El tiburón martillo tiene la cabeza lateralmente expandida con los ojos y la nariz en sus márgenes. Estudios indican que alteraciones de la forma de la cabeza de estos animales son adaptaciones que mejoran el desarrollo del sistema sensorial y estructuras asociadas a la visión binocular y otros órganos sensoriales. Dada la variedad de características, el objetivo del estudio fue describir la constitución anatómica del bulbo ocular. Se disecó el bulbo ocular junto a sus anexos. La descripción morfológica de las inserciones musculares y componentes del bulbo ocular se basaron en la observación directa, siendo éstas fotografiadas y catalogadas. Entre la esclera y la coroides se observó através de microscopía electrónica de barrido, una delgada capa supra-coroides, y vasos linfáticos junto a la matriz formada por tejidos conectivos. La coroides, era abundante en vasos sanguíneos y fibras de tejido conectivo. Se observaron escasas células pigmentarias llenas de melanina, lo que se asemeja a un anillo en forma de dedo, de manera que cubren la superficie de la esclera y células que contienen pigmentos. En la cápsula del lente, fue posible observar un recubrimiento celular, hialino y homogéneo. En la microscopía electrónica de barrido, se observó una cápsula muy gruesa, principalmente en la región frontal. La cápsula elástica,estaba constituida por láminas delgadas, principalmente, por fibras de colágeno. Se concluyó que el bulbo ocular de estos animales posee variaciones anatómicas relacionadas, principalmente, con la posición del bulbo ocular en el cráneo, túnica fibrosa y lente, lo que puede estar directamente relacionado con su hábitat.

Morphological study of the eye bulb of the hammerhead shark, sphyrna lewini (Elasmobranch: Carcharhinidae) / Estudio morfológico del bulbo ocular del tiburón martillo, sphyrna lewini (Elasmobranch: Carcharhinidae)

The hammerhead sharks shows a head laterally expanded with eyes and nostrils on the edges, which gives the species a hammer appearance. Another strand of studies indicates that the hypothesis of head shape alterations for better adaptions in the sensorial system with the development of structures associated to binocular sight and others sensorial organs. Given the variety of characteristics, the study aim was to describe the anatomical constitution of the hammerhead shark eye bulb. The bulb and its annexes exenteration was performed, and further dissection; the morphological description of the muscle insertions and eye bulb components were based on direct observation and were further photographed and catalogued. The eye bulb fragments and its annexes were histological technical. Between the sclera and the choroid it was possible to observe, by the electronical scanning microscopy, the thin layer called supra-choroid, in this region, spaces often filled by lymphatic vessels allied to a matrix formed by loose conjunctive tissue are found. In the choroid, a layer which is rich in blood vessels, loose conjunctive tissue and collagen fibers, was observed, besides pigmentary cells full of melanin in its interior, which result in its layer's dark color. Ciliary body is a choroid's dilatation; it has the aspect of a thick ring in finger-like shape, pigmented, covering the sclera surface and containing pigmentary cells. The crystalline capsule, which shows an acellular covering that, is hyaline and homogeneous. In the electronical scanning microscopy, it was observed that the capsule is extremely thick especially in the anterior face. The capsule is very elastic, constituted mainly by thin lamellae of collagenous fibers, as illustrated by the electronically scanning microscopy. Anatomic variations related mainly to the position of the eye bulb in the skull, fibrous tunica and lens call the attention and must be related to its habitat.

El tiburón martillo tiene la cabeza lateralmente expandida con los ojos y la nariz en sus márgenes. Estudios indican que alteraciones de la forma de la cabeza de estos animales son adaptaciones que mejoran el desarrollo del sistema sensorial y estructuras asociadas a la visión binocular y otros órganos sensoriales. Dada la variedad de características, el objetivo del estudio fue describir la constitución anatómica del bulbo ocular. Se disecó el bulbo ocular junto a sus anexos. La descripción morfológica de las inserciones musculares y componentes del bulbo ocular se basaron en la observación directa, siendo éstas fotografiadas y catalogadas. Entre la esclera y la coroides se observó através de microscopía electrónica de barrido, una delgada capa supra-coroides, y vasos linfáticos junto a la matriz formada por tejidos conectivos. La coroides, era abundante en vasos sanguíneos y fibras de tejido conectivo. Se observaron escasas células pigmentarias llenas de melanina, lo que se asemeja a un anillo en forma de dedo, de manera que cubren la superficie de la esclera y células que contienen pigmentos. En la cápsula del lente, fue posible observar un recubrimiento celular, hialino y homogéneo. En la microscopía electrónica de barrido, se observó una cápsula muy gruesa, principalmente en la región frontal. La cápsula elástica,estaba constituida por láminas delgadas, principalmente, por fibras de colágeno. Se concluyó que el bulbo ocular de estos animales posee variaciones anatómicas relacionadas, principalmente, con la posición del bulbo ocular en el cráneo, túnica fibrosa y lente, lo que puede estar directamente relacionado con su hábitat.

Adrenal glands of Spix's yellow-toothed cavy (Galea spixii, Wagler, 1831): morphological and morphometric aspects / Glândulas adrenais de preás (Galea spixii, Wagler, 1831): aspectos morfológicos e morfométricos

Abstract Considering the physiological importance and need of greater morphophysiological knowledge of adrenal glands, the aims of present study were compare the morphometric data between left and right adrenal of male and female; perform a histological, scanning and transmission electron microscopy study showing tissue constitution of glands; finally, in order to define the presence and correct site of the cytochrome P450c17 expression in adrenal glands, immunohistochemical study of this enzyme was performed in 18 adrenal glands (right n=9 and left n=9) of nine adult Galea spixii (four males and five females). Right adrenal was more cranially positioned than left adrenal; dimensions (weight, length and width) of right adrenal was larger than left adrenal; no differences between male and female body and adrenal measurements were found; the morphology of cells and different amounts of lipid droplets may be related to the different demands of steroid hormones production, related to each zone of the adrenal cortex; and, the cytochrome P450c17 immunolocalization in fasciculate and reticular zone may be related with synthesis of 17-hydroxy-pregnenolone, 17-hydroxy-progesterone, dehydroepiandrosterone or androstenedione.

Resumo Considerando a importância fisiológica e a necessidade de melhor conhecimento morfofisiológico das glândulas adrenais, os objetivos do presente estudo foram comparar dados morfométricos entre adrenais direitas e esquerdas de machos e fêmeas; realizar estudos histológicos e microscopia eletrônica de varredura e transmissão revelando a constituição tecidual das glândulas; finalmente, foi realizado estudo imunohistoquímico com objetivo de definir a presença e correto sítio de expressão da citocromo P450c17 nas glândulas adrenais em 18 glândulas adrenais (direita n=9 e esquerda n=9) de nove Galea spixii adultos (quatro machos e cinco fêmeas). A glândula adrenal direita estava posicionada mais cranialmente que a adrenal esquerda; as dimensões (peso, comprimento e largura) da glândula adrenal direita foram maiores que as da esquerda; não houve diferença entre as medidas corpóreas e das adrenais entre machos e fêmeas; a morfologia das células e diferentes quantidades de gotículas lipídicas podem estar relacionadas a diferentes demandas na produção de hormônios esteroides relacionados a cada zona do córtex adrenal; e, a imunolocalização da citocromo P450 na zona fasciculada e reticular pode estar relacionado com a síntese de 17-hidroxi-pregnenolona, 17-hidroxi-progesterona, dehidroepiandrosterona ou androstenediona.

Adrenal glands of Spix's yellow-toothed cavy (Galea spixii, Wagler, 1831): morphological and morphometric aspects.

Considering the physiological importance and need of greater morphophysiological knowledge of adrenal glands, the aims of present study were compare the morphometric data between left and right adrenal of male and female; perform a histological, scanning and transmission electron microscopy study showing tissue constitution of glands; finally, in order to define the presence and correct site of the cytochrome P450c17 expression in adrenal glands, immunohistochemical study of this enzyme was performed in 18 adrenal glands (right n=9 and left n=9) of nine adult Galea spixii (four males and five females). Right adrenal was more cranially positioned than left adrenal; dimensions (weight, length and width) of right adrenal was larger than left adrenal; no differences between male and female body and adrenal measurements were found; the morphology of cells and different amounts of lipid droplets may be related to the different demands of steroid hormones production, related to each zone of the adrenal cortex; and, the cytochrome P450c17 immunolocalization in fasciculate and reticular zone may be related with synthesis of 17-hydroxy-pregnenolone, 17-hydroxy-progesterone, dehydroepiandrosterone or androstenedione.

Morphological Aspects of Oral Denticles in the Sharpnose Shark Rhizoprionodon lalandii (Müller and Henle, 1839) (Elasmobranchii, Carcharhinidae).

The oral denticles of some elasmobranchs are found on the surface of the oral cavity and are homologous to those on the body surface, being well developed, independent and non-growing, with varying morphology and distribution depending on the species. The structural and three-dimensional characteristics of oral denticles from the rostro-ventral surface of the sharpnose shark Rhizoprionodon lalandii were described following imaging by both light and scanning electron microscopy. The light microscopy results showed that the triangular shape of the denticles consisted of a base and an apex. Picrosirius staining showed the arrangement of collagen fibres and oral denticles, and a predominance of type-I collagen was found in both structures under polarized light. There was a broad homogeneous distribution of denticles on the ventral surface, forming a leaf-like shape with the cusp facing the caudal region. Interlocking, hexagonal, geometric structures on its rostral side and ridges on the rostral side of the oral denticles were observed under increased magnification. We concluded that the denticle morphology found in R. lalandii differ of others analysed species, and the descriptions of these structures therefore provide important information for the classification of the species. In this species, the main functions can be assigned to help reduce hydrodynamic drag, particularly by this being a species that uses ram ventilation, and to protect the epithelium of the oropharynx of abrasion and parasites.

Ultrastructure of dermal denticles in sharpnose shark (Rhizoprionodon lalandii) (Elasmobranchii, Carcharhinidae).

The presence of denticles in the external surface, oral cavity, fins, and clasper of Elasmobranchii has been widely reported. These structures, called body denticles, may be observed on the body surface of sharks. Dermal and oral denticles are made up by a basal plate that is embedded in the dermis, forming a peduncle that grows from the base to the crown. These denticles may protect the skin against abrasion, and improve hydrodynamics and gill arches function. Rhizoprionodon lalandii is a widely distributed and very common species in Brazilian coastal areas. The aims of this study was to compare the morphology of oral and body denticles of R. lalandii to understand the implications of these structures in the behavior of these animals. Morphological analysis showed that there are differences between dermal and oral denticles, which are related to their role in different body regions. Body denticles have three cusps, and well-defined crests and ridges, and literature data suggest that suggest that hydrodynamics is their main function. Most of the oral denticles have only one cusp, and their morphology and distribution showed that their main functions are preparing food to be swallowed and protecting the oral cavity against abrasion.

Ultrastructure of spermatogenesis in Spix's yellow-toothed cavy (Galea spixii).

This was a pioneer study of the spermatogenic process from the onset of puberty in Spix's yellow-toothed cavies (SYC, Galea spixii) bred in captivity. The study aimed to characterize fine structure of spermatogenesis. Twelve testes from pubertal and post-pubertal SYC males were studied using transmission electron microscopy. Spermatogenesis can be divided into three phases: proliferation, meiosis, and spermiogenesis. In proliferation phase, three types of spermatogonia were identified and characterized as A(dark), A(pale), and B. In the second phase, spermatocytes (2n) undergo meiotic divisions that generate spermatids (n); the process begins in spermatocytes in the preleptotene stage when they increase their nuclear size, differentiating into spermatocytes in the leptotene stage when cell division is initiated. In addition, we found chromatin condensation, and formation of a structure composed of proteins that formed a central shaft and two lateral bars associated with pairing of homologous chromosomes. During spermiogenesis, the following main events occurred: condensation of nuclear chromatin, formation of acrosome with perfuratorium, elimination of residual cytoplasm, and development of the flagellum. The sperm head is different from that of other rodents. The endoplasmic reticulum and the Golgi complex are the two main organelles demonstrated during this process. These organelles collaborate through synthesis of proteins and hormones for the development of germ cells during spermatogenesis in SYC.

Morphological analysis of the tongue and the digestive tube of saffron finch (Sicalis flaveola brasiliensis, LINNAUES 1766) apprehended by CETAS/IBAMA-PB.

The saffron finch (Sicalis flaveola brasiliensis) is a Passeriforme of the Brazilian wildlife. There are scarcely any morphological studies on it, although it is frequently trafficked for its birdsong abilities. Its peculiarities, such as territorialism and developed syrinx that provides outstanding song, draw attention towards its domestication. Thus, this study aimed to morphologically describe the tongue and digestive tube organs of this species to furnish subsidies for nutritional, clinical and conservation studies. Forty-one birds from the Wild Animal Screening Center (CETAS)/Brazilian Institute of Environment (IBAMA)/city of Cabedelo, state of Paraíba (PB) were used. Samples were collected, identified and sent to standard light microscopy; samples of proventriculus and gizzard were sent to Scanning Electron Microscopy (SEM). The samples showed stratification similar to that of other domestic and wild birds, confirmed in the scanning electron microscopy; however, they differed in the absence of dermal papillae in the tongue, lack of ingluvial glands and lack of muscular mucosa and sub-mucosa in the large intestine.

Proliferation of fibroblasts and endothelial cells is enhanced by treatment with Phyllocaulis boraceiensis mucus.

Previously, mucus of some molluscs has been studied as a potential source of new natural compounds capable of inducing cell proliferation and of remodelling tissue. Here, the focus of the study is possible use of mucus released by Phyllocaulis boraceiensis - a compound inducing cell proliferation and enhancing collagen synthesis in dermal fibroblasts and inducing proliferation human endothelial cell cultures. Fibroblasts treated with P. boraceiensis mucus at concentrations below 0.012 µg/µl developed high rates of proliferation, as evaluated using MTT assay; the proliferative effect was dose-dependent. Production and secretion of extracellular matrix components and collagen type I fibres were enhanced after 24 h of treatment, revealing a hormesis effect, biphasic dose response - low dose for proliferation yet toxic at high dose. No significant change in proliferation was observed in treated endothelial cells and production of lipid polyunsaturated free radicals was low in both cell types. Treatment with P. boraceiensis mucus produced pronounced changes in fibroblast cell number and morphology, and in quantities of well-ordered collagen deposition. These results support the premise that Phyllocaulis boraceiensis mucus demonstrates proliferative properties in cells involved in the healing process.

Evaluation of the contribution of the placentomal fusion during gestation in cattle.

The placenta of mammals is a structure formed by the juxtaposition of the fetal membranes and the maternal tissues. The main function of the placenta is to regulate the physiological interchange between the fetus and the mother as well as to operate as an important endocrine organ during the gestation. The placentomal fusions were characterized throughout gestation of cattle using macroscopic, histological and flow cytometry analyses. Analyzing the cell cycle phases with a flow cytometry, a balance between the G2M phase and apoptosis was observed, suggesting that the placentomal fusions do not interfere in the placentary maturation process, which is a pre-requirement for the fetal-maternal disconnection and the release of fetal membrane.

Evaluation of the contribution of the placentomal fusion during gestation in cattle

The placenta of mammals is a structure formed by the juxtaposition of the fetal membranes and the maternal tissues. The main function of the placenta is to regulate the physiological interchange between the fetus and the mother as well as to operate as animportant endocrine organ during the gestation. The placentomal fusions were characterized throughout gestation of cattle using macroscopic, histological and flow cytometryanalyses. Analyzing the cell cycle phases with a flow cytometry, a balance between the G2M phase and apoptosis was observed, suggesting that the placentomal fusions do not interferein the placentary maturation process, which is a pre-requirement for the fetal-maternal disconnection and the release of fetal membrane.

Cell cycle and apoptosis in normal and cloned bovine near-term placentae.

The bovine maternal epithelium is composed of cuboidal cells interspersed with low columnar cells having centrally located nuclei. Bovine trophoblast is composed of two cell types: mononuclear trophoblastic and giant trophoblastic cells that can have two or more nuclei. Number of apoptotic cells and proliferative cells are variable in both cell populations. This study compared tissue growth and apoptosis by flow cytometry in the cell population found at distinct placental regions (central region of placentomes, < or =1-cm microplacentomes and the interplacentomal region) between normal and cloned near-term bovine pregnancies. After a morphological comparison between regions and groups (controls vs. clones), a lesser proportion of diploid to tetraploid cells was observed in the central region of placentomes and in microplacentomes from cloned-derived pregnancies. In addition, cloned animals had a fewer apoptotic cells in the central region of the placentome and in interplacentomal region and a greater proliferative capacity in all regions (cells in G(2)/M) near term as opposed to control animals. These results may reveal the existence of a relationship between such changes in the proportions of uterine and trophoblastic epithelial cells at the end of pregnancy and normal placental function. This could be related to faulty placentation in early pregnancy, placental insufficiency during pregnancy or lack of placental and/or fetal maturation in late pregnancy, which may contribute to some of the abnormalities after in vitro embryo manipulations, such as poor preparation and initiation of parturition, prolonged gestation and lesser post-natal survival in some cloned animals.