Tissue-engineered blood vessel substitute by reconstruction of endothelium using mesenchymal stem cells induced by platelet growth factors

Background: Cardiovascular diseases remain leaders as the major causes of mortality in Western society. Restoration of the circulation through construction of bypass surgical treatment is regarded as the gold standard treatment of peripheral vascular diseases, and grafts are necessary for this purpose. The great saphenous vein is often not available and synthetic grafts have their limitations. Therefore, new techniques to produce alternative grafts have been developed and, in this sense, tissue engineering is a promising alternative to provide biocompatible grafts. This study objective was to reconstruct the endothelium layer of decellularized vein scaffolds, using mesenchymal stem cells (MSCs) and growth factors obtained from platelets. Methods: Fifteen nonpregnant female adult rabbits were used for all experiments. Adipose tissue and vena cava were obtained and subjected to MSCs isolation and tissue decellularization, respectively. MSCs were subjected to differentiation using endothelial inductor growth factor (EIGF) obtained from human platelet lysates. Immunofluorescence, histological and immunohistochemical analyses were employed for the final characterization of the obtained blood vessel substitute. Results: The scaffolds were successfully decellularized with sodium dodecyl sulfate. MSCs actively adhered at the scaffolds, and through stimulation with EIGF were differentiated into functional endothelial cells, secreting significantly higher quantities of von Willebrand factor (0.85 μg/mL; P < .05) than cells cultivated under the same conditions, without EIGF (0.085 μg/mL). Cells with evident morphologic characteristics of endothelium were seen at the lumen of the scaffolds. These cells also stained positive for fascin protein, which is highly expressed by differentiated endothelial cells. Conclusions: Taken together, the use of decellularized bioscaffold and subcutaneous MSCs seems to be a potential approach to obtain bioengineered blood vessels, in the presence of EIGF supplementation. © 2013 Society for Vascular Surgery.

Ovarian blood vessel occlusion as a surgical sterilization method in rats

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Profile of cortisol, glycaemia, and blood parameters of American Bullfrog tadpoles Lithobates catesbeianus exposed to density and hypoxia stressors

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Light microscopy and computer three-dimensional reconstruction of the blood capillaries of the enamel organ of rat molar tooth germs

We performed a light microscope and a computer three-dimensional reconstruction study of serial sections of the molar enamel organ of 3- and 5-day-old rats perfused with Indian ink through the arterial system. The tooth germs were fixed in Bouin's solution, embedded in paraffin, sectioned and stained with haematoxylin and eosin. For the three-dimensional reconstruction, light micrographs of the serial sections were digitized, and aligned using the serial EM Align software downloaded from http://synapses.bu.edu/tools/. After alignment, the boundaries of the India-ink-filled blood vessels were manually traced with a mouse using the software IGL trace (version 1.26b), also downloaded from the above website. After tracing, a three-dimensional representation of the blood vessel contours was generated in a VRML format and visualized with the help of the software Cortona Web3D viewer (version 4.0) downloaded from http://www.parallelgraphics.com/products/cortona. Our results showed that in regions where ameloblasts are polarized the capillaries are arranged in three distinct levels: (1) penetrating and leaving capillaries in relation to the outer enamel epithelium; (2) capillaries crossing and branching inside the stellate reticulum; and (3) capillaries branching and anastomosing profusely within the stratum intermedium, thereby forming an extensive capillary plexus intimately associated with the cells of the stratum intermedium. The existence of a conspicuous capillary plexus intermingled with cells of the stratum intermedium, as shown in our results, suggests that some molecules produced by cells of the stratum intermedium could be released into the capillary plexus and thereafter carried to the dental follicle.

A Reproducible Venous Thrombosis Model in Horses Induced by the Combination of an Endothelial Lesion and Blood Flow Stasis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Contribution to the study of aortic mural structure of opossum (Didelphis albiventris)

Opossum is considered one of the most primitive mammals, with transition evolutive characteristics. In mammals, the aorta artery is referred as the main body blood vessel. The arteries wall structural organization follows a basic pattern, being contituted of three tunics: Intima, Median and Adventicial. After euthanasia, three samples of opossum had segments from the aorta artery ascendent, thoracic descending and abdominal descending portions removed, fixed in phormalin at 10% for 48 hours. Then, the material was washed in alcohol 70% several times, dehydrated in alcohois of growing concentrations, diafanized in xylol and included in "paraplast". Cuts with 5 to 7 mu m of thickness were placed in histological laminae and submitted to color methods of Hematoxilin-Eosin, Masson, Mallory and Calleja Tricromics. It was observed that in the different portions ascendent, thoracic and abdominal descending of aorta, the intima tunic presents much thick, made up of a layer of smooth muscular cells and elastic fibers, forming a limiting internal elastic membrane. In the three portions studied, the median tunic was the most evident layer, constituted of colagen fibers, smooth muscular cells arranged in a circular manner and elastic fibers, showing a variation in the mural elements proportion. It was evidenced the presence of a external elastic lamina, marking the transition between the median and adventicial tunic, formed by elastic fibers condensation. The aortic adventicial tunic showed to be little organized, having in its structure predominantely colagen fibers beans with some isolated smooth muscular fibers or in small fascicules among a few elastic fibers.

Calcaneal Tendon Regions Exhibit Different MMP-2 Activation After Vertical Jumping and Treadmill Running

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Aortic aneurysm surgery: problems and innovations

Um longo caminho foi percorrido desde as primeiras intervenções cirúrgicas feitas para o tratamento do aneurisma de aorta abdominal. Ao longo deste tempo, várias inovações foram criadas, no sentido de reduzir a invasividade dos procedimentos e melhorar sua segurança e durabilidade. Nesta revisão, são discutidos os principais e recentes avanços em intervenções sobre aneurismas aórticos, incluindo a restauração aórtica endovascular, a cirurgia aórtica videolaparoscópica, as técnicas híbridas convencionais e endovasculares, as técnicas combinadas videolaparoscópicas e endovasculares, bem como as perspectivas futuras, tanto para aorta torácica como abdominal. Diante de tantas transformações e evolução, o cirurgião vascular moderno terá que ter sua mente aberta para as novidades e desenvolver capacitação ampla com diferentes técnicas para proporcionar a melhor opção terapêutica para seus pacientes.

Placentation in cloned cattle: Structure and microvascular architecture

To elucidate the morphological differences between placentas from normal and cloned cattle pregnancies reaching term, the umbilical cord, placentomes and interplacentomal region of the fetal membranes were examined macroscopically as well as by light and scanning electron microscopy. In pregnancies established by somatic nucleus transfer (NT), the umbilical cord and fetal membranes were edematous. Placentomal fusion was common, resulting in increased size and a decreased number of placentomes. Extensive areas of the chorioallantoic membrane were devoid of placentomes. An increased number of functional or accessory microcotyledons (< 1 cm) were present at the maternally oriented surface of fetal membranes. Extensive areas of extravasated maternal blood were present within the placentomes and in the interplacentomal region. The crypts on the caruncular surface were dilated and accommodated complexes of more than one primary villus, as opposed to a single villus in non-cloned placentae. Scanning electron microscopy of blood vessel casts revealed that there was also more than one stem artery per villous tree and that the ramification of the vessels failed to form dense complexes of capillary loops and sinusoidal dilations as in normal pregnancies. At the materno-fetal interface, however, the trophoblast and uterine epithelium had normal histology. In conclusion, the NT placentas had a range of pathomorphological changes; this was likely associated with the poor clinical outcome of NT pregnancies. (c) 2007 Elsevier B.V. All rights reserved.

Sentinel node mapping in a pregnant woman with cervical cancer: a case report

The possibility of reducing morbidity associated with surgical dissection while maintaining accurate tumor staging is one of the greatest advantages of the sentinel node approach in surgical oncology. The sentinel node mapping has already proven to be useful in melanoma, breast cancer, and vulvar cancer. We report the first case of sentinel node detection by technetium-labeled radiocolloid in a pregnant woman with cervical cancer. The histologic analysis of the operative specimen showed a poorly differentiated squamous carcinoma with metastasis in the sentinel node and a neoplasic embolus in a blood vessel of the placental bed. The lymphatic mapping and sentinel lymph node detection are feasible during pregnancy.

Endogenous corticosteroids and insulin in acute inflammation

Carrageenin-induced inflammatory responses in the hindpaws of rats were quantitated by measuring: (1) alterations in volumes of the paws; and (2) alterations in concentration of dye, previously injected intravenously, which was recovered in perfusates from the paws. The inflammatory response in one paw was attenuated by previously inducing an inflammatory response in the contralateral paw. The effect was abolished by pretreatment with insulin. Indexes of adrenal activity were increased after the induction of the inflammatory response and they were not attenuated by pretreatment with insulin. Adrenal hyperactivity was characterized by increased serum corticosterone concentration, decreased adrenal ascorbic acid content, and reduced number of circulating eosinophils. It is concluded that inflammatory stimuli which lead to alterations in microvessels depend on a facilitatory effect of insulin. This effect is antagonized by glucocorticoids released in enhanced concentrations after the application of noxious stimuli. Therefore, endogenous insulin and glucocorticoids act as modulators of inflammatory responses.

Immunohistochemical characterisation of probable intravascular haematopoiesis in the vasa rectae of the renal medulla in acute tubular necrosis

Immunohistochemistry was applied to identify the nature of the nucleated cells that accumulate in the vasa rectae of the corticomedullary junction in acute tubular necrosis. In all 6 cases studied, there were intravascular cells that reacted with monoclonal antibodies to erythroblast, macrophages, myeloid cells, T and B lymphocytes and rare megakaryocytes. The findings are consistent with the occurrence of intravascular haematopoiesis in the renal medulia in acute tubular necrosis.