Proliferación y la viabilidad de las células derivadas del estroma mesenquimal de epiplón, el tejido adiposo perirrenal y el bolso de grasa interescapular en conejos (Oryctolagus cuniculus) / Proliferation and viability of mesenchymal stromal cells derived from omentum, fat perirenal and adipose interscapular pouch in rabbits (Oryctolagus cuniculus) / Proliferação e viabilidade de células-tronco derivadas do omento maior, tecido adiposo perirrenal e bolsa adiposa interescapular em coelhos (Oryctolagus cuniculus)

El tejido adiposo es una fuente alternativa de células madre mesenquimales, se presenta como el número de aumento de la colección de células menos invasiva y permitiendo comparación con el método de la médula ósea (MO). En los seres humanos, el metabolismo del tejido adiposo difiere de acuerdo con la localización anatómica. En ratones, se observaron diferencias en la composición celular y la capacidad de diferenciación de las células madre mesenquimales derivadas de tejido adiposo (ADSC), de acuerdo a las regiones anatómicas. El objetivo de este estudio es comparar la proliferación y la viabilidad de las células mesenquimales estromales multipotentes (CTMs) derivados de la bolsa de grasa interescapular, el epiplón mayor y la grasa perirrenal de conejos. Para esto, tres conejos fueron sometidos a cirugía para la extirpación de un tejido adiposo omental, perirrenal y interescapular abultada bolsa. Las células se procesaron y se cultivaron en una atmósfera humidificada con 5% de CO2, y posteriormente se tiñeron con azul de tripano y se contaron en una cámara de Neubauer. Llegamos a la conclusión de que hubo una variación en la capacidad proliferativa de CTMs con respecto a las regiones estudiadas, con la ventaja de la grasa omental en las otras dos regiones. La viabilidad de CTMs las tres regiones mostró variación durante los pasajes.(AU)

Adipose tissue is an alternative source of mesenchymal stem cell. It can be obtained by less invasive method and in larger quantities when compared with bone marrow. In humans, adipose tissue metabolism differs according to anatomic location. In mices, according to the anatomical regions, differences were observed in cellular composition and differentiation capacity of adipose-derived stromal cells (ADSCs). The objective of this study is to compare the viability and proliferation of mesenchymal stromal cells (MSC) derived from the bag interscapular fat, omental and perirenal fats of rabbits. For this, three rabbits were undergone to surgical collect fat from difference sites. Cells were processed and grown in a humidified atmosphere containing 5% CO2, and subsequently they were stained with Trypan blue and counted in a Neubauer chamber. It was concluded that there was variation in the proliferative capacity of MSCs over the regions studied, with advantage of great omental fat about the other regions. The viability of MSCs from the three regions showed variation during passages.(AU)

O tecido adiposo é fonte alternativa de células-tronco mesenquimais, apresentando-se como método menos invasivo e permitindo coleta de maior quantidade celular em comparação a medula óssea (MO). Nos humanos, o tecido adiposo apresenta diferenças de metabolismo, conforme a localização anatômica. Em camundongos, foram observadas diferenças em relação à composição celular e à capacidade de diferenciação das células tronco mesenquimais derivadas do tecido adiposo (ADSCs), de acordo com as regiões anatômicas. O objetivo deste trabalho é comparar a proliferação e viabilidade de células-tronco mesenquimais (CTMs) derivadas da bolsa adiposa interescapular, do omento maior e da gordura da região perirrenal de coelhos. Para isso, três coelhos foram submetidos a procedimentos cirúrgicos para remoção de tecido adiposo do omento maior, região perirrenal e bolsa adiposa interescapular. As células foram processadas e cultivadas em atmosfera umidificada a CO2 5%, sendo posteriormente coradas com Azul de Tripan e contadas em câmara de Neubauer. Concluí-se que houve variação na capacidade proliferativa das CTMs em relação as regiões estudadas, com vantagem da gordura omental sobre as outras duas regiões. A viabilidade das CTMs das três regiões apresentou variação durante as passagens.(AU)

Copaiba oil effect on experimental jaw defect in Wistar rats

PURPOSE: To evaluate the effects of copaiba oil on jaw defects repair in Wistar rats treated with bioglass or adipose tissue. METHODS: A jaw defect was randomly created in forty-two rats and filled with bioglass or adipose tissue. The two groups (Gbio and Gcell) were subdivided in three subgroups with seven animals each according to gavage administration: control (distillated water), oil (copaiba oil) and melox (meloxicam). Euthanasia was performed after forty post-operative days. The bone formation was analyzed regarding the histological aspects. RESULTS: The osteoclasts activity was observed only in four subgroups (p=0.78). Regarding the osteoblasts presence, it was very similar between the subgroups, the difference was due to Gcell-melox (p=0.009) that presented less osteoblastic activity. The inflammatory cells were more evident in Gcell-melox subgroup, however, there was no difference in comparison with the other subgroups (p=0.52). Bone formation was observed in all subgroups, just two animals showed no bone formation even after 40 days. More than 50% of bone matrix mineralization was observed in 56% (23 animals) of the analyzed areas. The bone matrix mineralization was not different between subgroups (p=0.60). CONCLUSIONS: The subgroups that received copaiba oil showed bone repair, although not statistically significant in comparison to subgroups treated whit meloxicam or controls. Copaiba oil administered by gavage had no effect on bone repair in this experimental model.(AU)

Experimental model of obtaining tissue adipose, mesenchymal stem cells isolation and distribution in surgery flaps in rats

PURPOSE: To investigate the experimental model for obtaining adipose tissue, isolation, characterization of mesenchymal stem cells and evaluation of their distribution in the tram flap in rats. METHODS: Five rats of Wistar were randomly assigned to two groups. In group I, three animals underwent removal of adipose tissue in the groin procedure to establish the experimental model and obtain a cell lineage. The animals of group II (n = 2) underwent surgical flap procedure, and satisfaction injection of mesenchymal stem cells pretreated with marker fluorescente. RESULTS: obtaining adipose tissue of the inguinal region of the rat proved to be possible. The isolated cells were characterized as mesenchymal stem cells and fluorescence microscopy showed the presence of multiple cells arranged around blood vessels and capillaries. CONCLUSION: It was possible to establish an experimental model for obtaining adipose tissue for isolation of mesenchymal stem cells and their distribution in the TRAM flap in rats.(AU)

A new species of Hyphessobrycon Durbin (Characiformes: Characidae) from the middle rio São Francisco and upper and middle rio Tocantins basins, Brazil, with comments on its biogeographic history

A new species of Hyphessobrycon Durbin is described from tributaries of the rio Grande (rio São Francisco basin) draining east to the Serra Geral de Goiás, of the rio São Domingos (upper rio Tocantins basin) and rio do Sono basins (middle rio Tocantins basin). Hyphessobrycon diastatos can be diagnosed from its congeners by the absence of humeral spot, 15-18 branched anal-fin rays, 1-3 maxillary conical to tricuspid teeth, a relatively well-defined dark caudal-peduncle spot, and elongation of dorsal- and anal-fin rays in mature males. The biogeographical significance of the distribution of the new species is evaluated and discussed.(AU)

Uma espécie nova de Hyphessobrycon Durbin é descrita dos tributários do rio Grande (bacia do rio São Francisco) que drenam a leste a Serra Geral de Goiás, das bacias dos rios São Domingos (bacia do alto rio Tocantins) e rio do Sono (bacia do médio rio Tocantins). Hyphessobrycon diastatos pode ser diagnosticada de seus congêneres pela ausência de mancha umeral, 15-18 raios ramificados na nadadeira anal, 1-3 dentes maxilares cônicos ou tricuspidados, uma mancha escura relativamente bem definida no pedúnculo caudal e alongamento das nadadeiras dorsal e anal em machos maduros. O significado biogeográfico da distribuição da espécie nova é avaliado e discutido.(AU)

Adipose-derived stem cells (ADSC) in the viability of a random pattern dorsal skin flap in rats

To evaluate the viability of random pattern dorsal skin flaps in rats after injection of adipose-derived stem cells (ADSC). Thirty five adult male Wistar EPM rats (weight 250-300 g) were distributed, at random, in two groups. I- Control (flap elevation with injection of saline solution) with fifteen animals and II- Experimental (flap elevation with injection of ADSC ) with fifteen animal. The ADSC were isolated from others five adult male rats. A dorsal skin flap measuring 10x4 cm was raised and a plastic barrier was placed between the flap and its bed in both groups and the injection (cells or saline solution) were perfomed immediately after the surgery. The percentage of flap necrosis was measured on the seventh postoperative day. The ADSC were able to replicate in our culture conditions. We also induced their adipogenic, osteogenic and chondrogenic differentiation to verify their mesenchymal stem cells potentiality in vitro. The results were statistically significant showing that the ADSC decreased the area of necrosis (p<0.05). CONCLUSIONS: The cells demonstrated adipogenic, osteogenic and chondrogenic differentiation potential in vitro. The administration of adipose-derived stem cells was effective to increase the viability of the random random pattern dorsal skin flaps in rats.(AU)

Adipose-derived stem cells (ADSC) in the viability of random skin flap in rats

To evaluate the effects of the adipose-derived stem cells (ADSC) in the viability of random skin flap in rats. Thirty five adult male Wistar rats (weight 250-300 g) were used. ADSC were isolated from adult male rats (n=5). ADSC were separated, cultured and then analyzed. A dorsal skin flap measuring 10x4 cm was raised and a plastic barrier was placed between the flap and its bed. After the surgical procedure, the animals were randomized into two groups (n=15 each group), group control and group ADSC. In all groups the procedures were performed immediately after the surgery. The percentage of flap necrosis was measured on the seventh postoperative day. The ADSC were able to replicate in our culture conditions. We also induced their adipogenic, osteogenic and chondrogenic differentiation, verifying their mesenchymal stem cells potentiality in vitro. The results were statistically significant showing that the ADSC decreased the area of necrosis (p<0.05). CONCLUSION: The cells demonstrated adipogenic, osteogenic and chondrogenic differentiation potential in vitro. The administration of adipose-derived stem cells was effective to increase the viability of the random skin flaps in rats.(AU)

Isolamento e caracterização de células mesenquimais do tecido adiposo de cães / Isolation and characterization of canine adipose-derived mesenchymal stem cells

As células-tronco mesenquimais (CTMs) diferenciam-se em várias linhagens e têm potencial de utilização na medicina regenerativa. As CTMs podem ser isoladas de vários tecidos de animais adultos. O objetivo deste estudo foi o isolamento das CTMs do tecido adiposo de cães, seu cultivo e diferenciação. Foram coletadas amostras de tecido adiposo subcutâneo de cinco cães. As CTMs foram isoladas, obtendo-se 146.803 (±49.533) células/g, cultivadas e diferenciadas em osteoblastos, adipócitos e condrócitos. Avaliaram-se a cinética do crescimento, a morfologia e a viabilidade celular. A caracterização citoquímica comprovou a natureza mesenquimal das células isoladas. O cultivo foi iniciado com 20.000 células/mL, verificando-se crescimento rápido até 72 horas (220.000 células/mL), fase exponencial entre 72 e 192 horas (455.000 células/mL), seguida de platô por saturação da densidade com 240 horas (355.000 células/mL). A viabilidade celular variou entre 96 e 100%. As CTMs em cultivo são fibroblásticas, fusiformes, com citoplasma basofílico e núcleo esférico. O comprimento médio das células variou entre 80,85 e 98,36µm, a largura média entre 17,40 e 28,79µm e o diâmetro médio do núcleo entre 15,46 e 17,74µm.(AU)

The applications of mesenchymal stem cells (MSCs) are becoming increasingly more promising for regenerative medicine and tissue engineering fields. MSCs can be isolated from adult animals from a variety of tissues, such as the adipose. This study focused on the isolation, culture and differentiation of MSCs from canine adipose tissue. Samples of subcutaneous adipose tissue from five dogs were collected. These cells were isolated, cultured and differentiated into osteoblasts, adipocytes and chondrocytes. We obtained 146,803 (±49,533) cells/g. Growth kinetics and viability studies were conducted during cell culture and the evaluation of cell differentiation was successfully performed by cytochemistry. The cell cultures were initiated with 20,000 MSCs/ml. Rapid growth was observed at 72 hours (220,000 cells/ml), the exponential phase between 72 and 192 hours (455,000 cells/ml) and saturation at 240 hours (355,000 cells/ml). The cellular viability ranged from 96 to 100%. MSCs in culture are fibroblastic cells, fusiform with basophilic cytoplasm and spherical nucleus. The length and width means of the cells and nuclear diameter ranged from 80.85-98.36µm, 17.40-28.79µm and 15.46-17.74µm respectively.(AU)

Evaluation of the donor site after liposuction with a syringe in rabbits / Avaliação da zona doadora após lipoaspiração com seringa em coelhos

PURPOSE: To evaluate the donor site of adipocytes as well as histopathological alterations secondary to liposuction. METHODS: All animals underwent liposuction with a syringe on the right side of the back. While the left side of the back was used as control and did not undergo intervention. The 10 rabbits were divided into two groups A and B according the postoperative day which were submitted to euthanasia: 90 and 120 days. All adipose tissue from the donor site was analyzed and compared with the control macroscopic and light microscopy. Tissues were weighed and analyzed searching for histological changes and late inflammatory response to trauma such as fibrosis, fat necrosis and inflammation and macrophage infiltration. RESULTS: There was wide variation in adipose tissue volume between the experimental and the control on macroscopic analysis. The presence of histopathological changes was found in two samples at 90 days. CONCLUSIONS: There was a relationship between the presence of fibrosis with the weight and number of days after liposuction surgery in rabbits. The study show macroscopic difference between control and experiment sides in all rabbits.(AU)

OBJETIVO: Avaliar a zona doadora de adipócitos bem como as alterações histopatológicas secundárias a lipoaspiração. MÉTODOS: Todos os animais foram submetidos à lipoaspiração com seringa no lado direito do dorso. Enquanto o lado esquerdo do dorso foi usado como controle, não sofrendo intervenção. Os 10 coelhos foram divididos em dois grupos A e B conforme o dia pós-operatório (DPO) os quais foram submetidos à eutanásia: 90 DPO e 120 DPO. Todo tecido adiposo da zona doadora foi analisado e comparados com o controle macroscopicamente e à microscopia óptica. Os tecidos foram pesados e foram analisadas as variações histológicas em busca de uma resposta inflamatória tardia ao trauma como fibrose, inflamação, necrose gordurosa e infiltrado macrofágico. RESULTADOS: Foi observada grande variação de volume de tecido adiposo entre o experimento e o controle do dorso a macroscópica e a presença de alteração histopatológica em duas amostras aos 90 dias. CONCLUSÕES: Houve relação entre a presença de fibrose com o peso e numero de dias pós-operatório na lipoaspiração de coelhos. O estudo mostrou diferença entre os lados experimento e controle em todos os coelhos.(AU)

Engraftment of human adipose derived stem cells delivered in a hyaluronic acid preparation in mice / Implante de células tronco do tecido adiposo humano numa preparação de ácido hialurônico em camundongos

PURPOSE: To evaluate the implant of human adipose derived stem cells (ADSC) delivered in hyaluronic acid gel (HA), injected in the subcutaneous of athymic mice. METHODS: Control implants -HA plus culture media was injected in the subcutaneous of the left sub scapular area of 12 athymic mice. ADSC implants: HA plus ADSC suspended in culture media was injected in the subcutaneous, at the contra lateral area, of the same animals. With eight weeks, animals were sacrificed and the recovered implants were processed for extraction of genomic DNA, and histological study by hematoxilin-eosin staining and immunufluorescence using anti human vimentin and anti von Willebrand factor antibodies. RESULTS: Controls: Not visualized at the injection site. An amorphous substance was observed in hematoxilin-eosin stained sections. Human vimentin and anti von Willebrand factor were not detected. No human DNA was detected. ADSC implants - A plug was visible at the site of injection. Fusiform cells were observed in sections stained by hematoxilin- eosin and both human vimentin and anti von Willebrand factor were detected by immunofluorescence. The presence of human DNA was confirmed. CONCLUSION: The delivery of human adipose derived stem cells in preparations of hyaluronic acid assured cells engraftment at the site of injection.(AU)

OBJETIVO: Avaliar o implante de células tronco do tecido adiposo humano (CTTAH) em gel de ácido hialurônico (AH), injetados no tecido subcutâneo de camundongos atímicos. MÉTODOS: Implantes controle - HA com meio de cultura foram injetados no tecido subcutâneo da região infraescapular esquerda de 12 camundongos atímicos. Implantes de CTTAH: HA com CTTAH suspensas em meio de cultura foi injetado no subcutâneo da região contra lateral, dos mesmos animais. Com oito semanas, os animais foram sacrificados e os implantes recuperados foram processados para extração de DNA genômico, estudo histológico por coloração por hematoxilina eosina e imnuoflurescência utilizando anticorpos anti vimentina humana e anti fator de von Willebrand. RESULTADOS: Controles - implantes não visualizados no local da injeção. Uma substância amorfa foi observada nos cortes corados por hematoxilina eosina. Vimentina humana e fator anti von Willebrand não foram identificados. DNA humano não foi detectado. Implantes de CTTAH - Uma massa era visível no local da injeção. Células fusiformes foram observadas nos corte corados com hematoxilina eosina. Tanto vimentina humana quanto fator de von Willebrand foram identificados pela imunofluorescência. A presença de DNA humano foi confirmada. CONCLUSÃO: O implante de células tronco do tecido adiposo humano em veículo de ácido hialurônico gel assegurou a manutenção das células no local do implante.(AU)