PURPOSE:There is a growing scientific interest in the plasticity and therapeutic potential of adipose-derived stem cells (ASCs), which are multipotent and abundant in adipose tissue and can differentiate in vitro into multiple lineages, including adipocytes, chondrocytes, osteoblasts, neural cells, endothelial cells and cardiomyocytes. The aim of this study was to isolate, cultivate and identify ASCs. METHODS: Human adipose precursor cells were obtained from subcutaneous abdominal tissue. Recently dispersed cells were separated by density centrifugation gradient, cultured and then analyzed. RESULTS: Human ASCs were able to replicate in our culture conditions. The cells maintained their phenotypes throughout the studied period on different passages confirming they suitability for in vitro cultivation. We also induced their adipogenic, osteogenic and chondrogenic differentiation, verifying their mesenchymal stem cells potentiality in vitro. Flow cytometry results showed that these cells expressed CD73, CD90 and CD105, (mesenchymal stem-cells markers), contrasting with the lack of expression of CD16, CD34 and CD45 (hematopoietic cells markers). RESUMO OBJETIVO:Há um interesse científico crescente na plasticidade e potencial terapêutico das células-tronco do tecido adiposo humano, células multipotentes e abundantes no tecido adiposo que podem se diferenciar in vitro em múltiplas linhagens celulares, incluindo adipócitos, condrócitos, osteoblastos, células neurais, endoteliais e cardiomiócitos. O objetivo deste estudo foi isolar, cultivar e identificar células-tronco do tecido adiposo humano. MÉTODOS: Células precursoras humanas do tecido adiposo foram obtidas de tecido abdominal subcutâneo. As células recém-dispersas foram separadas por gradiente de centrifugação por densidade, cultivadas e então analisadas. RESULTADOS: As células-tronco do tecido adiposo humano foram capazes de se replicar nas nossas condições de cultivo e mantiveram seu fenótipo em diferentes passagens durante o estudo, confirmando sua adequabilidade para cultivo in vitro. A diferenciação adipogênica, osteogênica e condrogênica também foi induzida, confirmando seu potencial de células-tronco mesenquimais in vitro. Os resultados de citometria de fluxo evidenciaram a expressão dos marcadores de células-tronco mesenquimais CD73, CD90 e CD105, contrastando com a falta de expressão dos marcadores de células hematopoiéticas CD16, CD34 e CD45. CONCLUSÃO: Foi possível isolar células-tronco do tecido adiposo humano por cultivo in vitro sem indução adipogênica, mantendo sua integridade funcional e altos níveis de proliferação. As células demonstraram potencial de diferenciação adipogênico, osteogênico e condrogênico in vitro. Descritores: Adipócitos. Células-Tronco Adultas. Células-Tronco Mesenquimais. Células-Tronco. Tecido Adiposo.Gaiba S et al.
PURPOSE:To propose an experimental burn model in NIH-3T3 cell line. METHODS:Induction of thermal injury in cultures of mouse fibroblast -NIH-3T3-cell line and determination of cell viability by MTT and imunofluorescence. RESULTS:The heating of the Petri dish increased proportionally to the temperature of the base and the time of exposure to microwave.In this in vitro burn model, using the cell line NIH-3T3 was observed drastic cellular injury with significant changes in cell viability and activity. It showed drastically modified cell morphology with altered membrane, cytoskeleton and nucleus, and low cellularity compared to the control group. CONCLUSION:The burn model in vitro using the cell line NIH-3T3 was reproductive and efficient. This burn model was possible to determine significant changes in cell activity and decreased viability, with drastic change in morphology, cell lysis and death.
PURPOSE:To evaluate the viability of random pattern dorsal skin flaps in rats after injection of adipose-derived stem cells (ADSC). METHODS: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. RESULTS: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.
Purpose: To evaluate the effects of the adipose-derived stem cells (ADSC) in the viability of random skin flap in rats.Methods: 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. Results: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.
RESULTS:Our results showed many secretory canals in resin fruit and leaf; identifying lipids, starch, lignin and phenolic compounds.The leaf extract induced genotoxicity and apoptosis in B16-F10 cells, since the fragmentation of DNA in the comet assay. The ethanolic extract of mangosteen obtained in the resin, leaf and fruit showed antimicrobial activity against Staphylococcus aureus and Escherichia coli with a MIC at 0.1 mg/mL. CONCLUSION:In conclusion, we have demonstrated both antimicrobial and antitumor activity of ethanol extract of mangosteen emphasizing its therapeutic potential in infectious diseases and in cancer, such as melanoma.
Objective. The aim of this study was to evaluate the bone healing after the usage of a scaffold enriched with bone marrow. Study Design. Ten rabbits were divided into 2 groups of 5 animals. Bilateral 12 mm diameter defects were created in the parietal bones. In control group Bio-Oss were inserted in both defects and, in experimental group, Bio-Oss enriched with autologous bone marrow were inserted in both defects. In these two groups, one of the calvarial defects was covered with Bio-Gide. The rabbits were sacrified 8 weeks after surgery and both CT and histomorphometric analysis were done. Results. The CT showed a lower remaining defect area in the experimental group covered with Bio-Gide when compared with control group, with and without Bio-Gide. The histomorphometrics showed no difference between groups regarding the non-vital mineralized tissue area. For vital mineralized tissue area, the experimental group covered with Bio-Gide obtained a higher percentage area when compared with control group, with and without Bio-Gide. For non-mineralized tissue area, the experimental group covered with Bio-Gide obtained a lower percentage area when compared with control group, with and without Bio-Gide. Conclusion. Both autologous bone marrow and membrane can contribute to the enhancement of bone healing.
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