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Id: biblio-950875
Autor: Cheng, Ming-Jun; Cao, Yun-Gui.
Título: TMPYP4 exerted antitumor effects in human cervical cancer cells through activation of p38 mitogen-activated protein kinase
Fonte: Biol. Res;50:24, 2017. graf.
Idioma: en.
Projeto: National Natural Science Foundation of China; . Chongqing Science and Technology.
Resumo: BACKGROUND: The aim of the present study was to investigate the potential effects of the 5,10,15,20-tetrakis (1-methylpyridinium-4-yl) porphyrin (TMPyP4) on the proliferation and apoptosis of human cervical cancer cells and the underlying mechanisms by which TMPyP4 exerted its actions. RESULTS: After human cervical cancer cells were treated with different doses of TMPyP4, cell viability was determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) method, the apoptosis was observed by flow cytometry (FCM), and the expression of p38 mitogen-activated protein kinase (MAPK), phosphated p38 MAPK (p-p38 MAPK), capase-3, MAPKAPK2 (MK-2) and poly ADP-ribose polymerase (PARP) was measured by Western blot analysis. The analysis revealed that TMPyP4 potently suppressed cell viability and induced the apoptosis of human cervical cancer cells in a dose-dependent manner. In addition, the up-regulation of p-p38 MAPK expression levels was detected in TMPyP4-treated human cervical cancer cells. However, followed by the block of p38 MAPK signaling pathway using the inhibitor SB203580, the effects of TMPyP4 on proliferation and apoptosis of human cervical cancer cells were significantly changed. CONCLUSIONS: It was indicated that TMPyP4-inhibited proliferation and -induced apoptosis in human cervical cancer cells was accompanied by activating the p38 MAPK signaling pathway. Taken together, our study demonstrates that TMPyP4 may represent a potential therapeutic method for the treatment of cervical carcinoma.
Descritores: Porfirinas/farmacologia
Neoplasias do Colo do Útero/tratamento farmacológico
Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
Antineoplásicos/farmacologia
-Sais de Tetrazólio
Células HeLa/efeitos dos fármacos
Sobrevivência Celular/efeitos dos fármacos
Células Cultivadas
Western Blotting
Reprodutibilidade dos Testes
Apoptose/efeitos dos fármacos
Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores
Proliferação de Células/efeitos dos fármacos
Ativação Enzimática/efeitos dos fármacos
Caspase 3/análise
Formazans
Limites: Humanos
Feminino
Tipo de Publ: Estudo de Avaliação
Responsável: CL1.1 - Biblioteca Central


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Id: lil-787334
Autor: Covre, Joyce L; Cristovam, Priscila C; Loureiro, Renata R; Hazarbassanov, Rossen M; Campos, Mauro; Sato, Élcio H; Gomes, José Álvaro P.
Título: The effects of riboflavin and ultraviolet light on keratocytes cultured in vitro / Avaliação do efeito da riboflavina e luz ultravioleta sobre os ceratócitos in vitro
Fonte: Arq. bras. oftalmol;79(3):180-185graf.
Idioma: en.
Resumo: ABSTRACT Purpose: To culture quiescent human keratocytes and evaluate the effects of ultraviolet light and riboflavin on human corneal keratocytes in vitro. Methods: Keratocytes were obtained from remaining corneoscleral ring donor corneas previously used in corneal transplant surgeries and cultured in DMEM/F12 with 2% FBS until confluence. Characterization of cultured cells was performed by immunofluorescence analysis for anti-cytokeratin-3, anti-Thy-1, anti-α-smooth muscle actin, and anti-lumican. Immunofluorescence was performed before and after treatment of cultured cells with either ultraviolet light or riboflavin. Corneal stromal cells were covered with collagen (200 µL or 500 µL) and 0.1% riboflavin, and then exposed to ultraviolet light at 370 nm for 30 minutes. After 24 hours, cytotoxicity was determined using MTT colorimetric assays, whereas cell viability was assessed using Hoechst 33342 and propidium iodide. Results: Cell cultures achieved confluence in approximately 20 days. Expression of the lumican was high, whereas no expression of CK3, Thy-1, and α-SMA was observed. After crosslinking, MTT colorimetric assays demonstrated a low toxicity rate, whereas Hoechst 33342/propidium iodide staining demonstrated a low rate of apoptosis and necrosis, respectively, in all collagen-treatment groups. Conclusion: Keratocytes can be successfully cultured in vitro and characterized by immunofluorescence using lumican. MTT colorimetric assays, and Hoechst 33342, and propidium iodide staining demonstrated a higher rate of cell death in cells cultured without collagen, indicating collagen protects keratocytes from the cytotoxic effects of ultraviolet light.

RESUMO Objetivo: Avaliar o efeito da aplicação da luz ultravioleta e riboflavina sobre ceratócitos da córnea humana in vitro. Métodos: Os ceratócitos foram obtidos a partir das rimas corneoesclerais remanescentes da trepanação de córneas previamente utilizadas em cirurgias de transplante de córnea e cultivadas em meio DMEM/F12 com 2% de FBS até atingir confluência. As culturas de células foram caracterizadas por imunofluorescência com os anticorpos K3 (marcador de células epiteliais), Thy-1 (marcador de fibroblasto) SMA (marcador de miofibroblasto) e Lumican (marcador de ceratócitos). Imunofluorescência também foi feita após o tratamento. As células do estroma da córnea foram cobertas com colágeno (200 µL e 500 µL) e 0,1% de riboflavina e exposta a luz UVA a 370 nm por 30 minutos. Após 24 horas, citotoxicidade foi determinada por ensaio de MTT e a viabilidade celular foi feita por Hoechst 33342/Iodeto de propideo. Resultados: As culturas de células atingiram confluência em aproximadamente 20 dias. Imunofluorescência apontou alta expressão para o marcador de ceratócitos (Lumican) e expressão negativa par os marcadores de células epiteliais (K3), fibroblasto (Thy-1) e miofibroblasto (α-SMA). Após o cross linking a análise de MTT mostrou baixa taxa de toxicidade e com a coloração de Hoechst 33342/Iodeto de propideo baixa taxa de apoptose e necrose respectivamente em todos os grupos que continham colágeno. Conclusão: As culturas de ceratócitos foram obtidas e caracterizadas por imunofluorescência através do marcador Lumican com sucesso. O ensaio de MTT e a coloração por Hoechst 33342 e iodeto de propídio, apresentaram maior índice de morte celular nos grupos que não continham colágeno, provando que protege as células contra os efeitos da luz UVA.
Descritores: Riboflavina/farmacologia
Raios Ultravioleta
Fármacos Fotossensibilizantes/farmacologia
Ceratócitos da Córnea/efeitos dos fármacos
Ceratócitos da Córnea/efeitos da radiação
-Sobrevivência Celular/efeitos dos fármacos
Sobrevivência Celular/efeitos da radiação
Células Cultivadas
Análise de Variância
Imunofluorescência
Colágeno/farmacologia
Apoptose/efeitos dos fármacos
Apoptose/efeitos da radiação
Substância Própria/citologia
Reagentes para Ligações Cruzadas/farmacologia
Fibroblastos/efeitos dos fármacos
Fibroblastos/efeitos da radiação
Formazans
Necrose
Limites: Humanos
Tipo de Publ: Estudo de Avaliação
Responsável: BR1.1 - BIREME


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Id: biblio-1089334
Autor: Gutiérrez-Barbosa, Hernando; Castañeda, Nadia Y; Castellanos, Jaime E.
Título: Differential replicative fitness of the four dengue virus serotypes circulating in Colombia in human liver Huh7 cells
Fonte: Braz. j. infect. dis;24(1):13-24, Feb. 2020. tab, graf.
Idioma: en.
Resumo: ABSTRACT Dengue has been a significant public health problem in Colombia since the simultaneous circulation of the four dengue virus serotypes. The replicative fitness of dengue is a biological feature important for virus evolution and contributes to elucidating the behavior of virus populations and viral pathogenesis. However, it has not yet been studied in Colombian isolates. This study aimed to compare the replicative fitness of the four dengue virus serotypes and understand the association between the serotypes, their in vitro infection ability, and their replication in target cells. We used three isolates of each DENV serotype to infect Huh-7 cells at an MOI of 0.5. The percentage of infected cells was evaluated by flow cytometry, cell viability was evaluated by MTT assay, and the pathogenicity index was calculated as a ratio of both parameters. The replicative fitness was measured by the number of viral genome copies produced using quantitative PCR and the production of infectious viral progeny was measured by plaque assay. We showed that Huh-7 cells were susceptible to infection with all the different strain isolates. Nevertheless, the biological characteristics, such as infectious ability and cell viability, were strain-dependent. We also found different degrees of pathogenicity between strains of the four serotypes, representative of the heterogeneity displayed in the circulating population. When we analyzed the replicative fitness using the mean values obtained from RT-qPCR and plaque assay for the different strains, we found serotype-dependent behavior. The highest mean values of replicative fitness were obtained for DENV-1 (log 4.9 PFU/ml) and DENV-4 (log 5.28 PFU/ml), followed by DENV-2 (log 3.9 PFU/ml) and DENV-3 (log 4.31 PFU/ml). The internal heterogeneity of the replicative fitness within each serotype could explain the simultaneous circulation of the four DENV serotypes in Colombia.
Descritores: Replicação Viral/genética
Vírus da Dengue/genética
Vírus da Dengue/patogenicidade
Sorogrupo
-Ensaio de Placa Viral
Valores de Referência
Sais de Tetrazólio
Fatores de Tempo
RNA Viral/genética
Linhagem Celular
Sobrevivência Celular
Células Cultivadas
Colômbia
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Citometria de Fluxo
Formazans
Fígado/citologia
Limites: Humanos
Responsável: BR1.1 - BIREME


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Id: biblio-886638
Autor: GÓMEZ-CANSINO, ROCIO; GUZMÁN-GUTIÉRREZ, SILVIA LAURA; CAMPOS-LARA, MARÍA GUADALUPE; ESPITIA-PINZÓN, CLARA INES; REYES-CHILPA, RICARDO.
Título: Natural Compounds from Mexican Medicinal Plants as Potential Drug Leads for Anti-Tuberculosis Drugs
Fonte: An. acad. bras. ciênc;89(1):31-43, Jan,-Mar. 2017. graf.
Idioma: en.
Projeto: UNAM and CONACyT.
Resumo: ABSTRACT In Mexican Traditional Medicine 187 plant species are used in the treatment of respiratory conditions that may be associated with tuberculosis. In this contribution, we review the ethnobotany, chemistry and pharmacology of 63 species whose extracts have been assayed for antimycobacterial activity in vitro. Among these, the most potent is Aristolochia brevipes (MIC= 12.5 µg/mL), followed by Aristolochia taliscana, Citrus sinensis, Chrysactinia mexicana, Persea americana, and Olea europaea (MIC<64 µg/mL). Other potent extracts (inhibition > 95%, 50 µg/mL) include: Amphipterygium adstringens, Larrea divaricata, and Phoradendron robinsoni. Several active compounds have been identified, the most potent are: Licarin A (isolated from A. taliscana), and 9-amino-9-methoxy-3,4-dihydro-2H-benzo[h]-chromen-2-one (transformation product of 9-methoxytariacuripyrone isolated from Aristolochia brevipes), both with MIC= 3.125 µg/mL, that is 8-fold less potent than the reference drug Rifampicin (MIC= 0.5 µg/mL). Any of the compounds or extracts here reviewed has been studied in clinical trials or with animal models; however, these should be accomplished since several are active against strains resistant to common drugs.
Descritores: Plantas Medicinais/química
Extratos Vegetais/farmacologia
Extratos Vegetais/química
Antituberculosos/farmacologia
Antituberculosos/química
-Sais de Tetrazólio
Contagem de Colônia Microbiana
Testes de Sensibilidade Microbiana
Reprodutibilidade dos Testes
Etnobotânica
Formazans
México
Mycobacterium tuberculosis/efeitos dos fármacos
Responsável: BR1.1 - BIREME


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Id: biblio-888968
Autor: Wang, WJ; Cheng, MH; Lin, JH; Weng, CS.
Título: Effect of a rosmarinic acid supplemented hemodialysis fluid on inflammation of human vascular endothelial cells
Fonte: Braz. j. med. biol. res = Rev. bras. pesqui. méd. biol;50(12):e6145, 2017. tab, graf.
Idioma: en.
Resumo: Chronic systemic inflammation and repetitive damage of vascular endothelia by incompatible dialysis system are probable causes of cardiovascular disease in patients on dialysis. The present study aimed to assess in vitro biocompatibility and anti-inflammatory effect of hemodialysis fluid supplemented with rosmarinic acid (RA) using human umbilical vein endothelial cells (HUVEC). HUVECs (5×106 cells/mL) were pre-exposed to 1 μg/mL of lipopolysaccharides (LPS) and incubated with RA-supplemented hemodialysis fluid (HDF). Cytotoxicity was assessed qualitatively by morphologic assessment and quantitatively by MTT assay. Expressions of proinflammatory mediators were assessed using quantitative real-time PCR and production of NO was quantified. Phosphorylation of AKT and nuclear localization of nuclear factor kappa B (NF-κB) were examined using western blotting. Exposure of HUVECs to RA-supplemented HDF had no influence on morphology and viability. Inhibition of proinflammatory mediator production in HUVECs by RA supplementation to HDF was significant in a dose-dependent manner. Exposure to RA-supplemented HDF resulted in a decrease in nitric oxide synthase expression and reduction of NO production in LPS-stimulated HUVECs. RA supplementation of HDF suppressed Akt activation in LPS-stimulated HUVECs. In addition, the level of cellular IκB was increased in parallel to a reduced nuclear translocation of NF-κB in LPS-induced endothelial cells. Our results suggest that RA-supplemented HDF is biocompatible and significantly suppressed inflammation induced in endothelial cells. In this respect, the use of HDF supplemented with RA could alleviate inflammation and improve long-term treatment of patients with renal failure on dialysis. Further clinical studies are required to confirm the effects.
Descritores: Anti-Inflamatórios não Esteroides/farmacologia
Materiais Biocompatíveis/farmacologia
Cinamatos/farmacologia
Depsídeos/farmacologia
Soluções para Hemodiálise/farmacologia
Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos
Inflamação/tratamento farmacológico
-Análise de Variância
Sobrevivência Celular/efeitos dos fármacos
Células Cultivadas
Citocinas/análise
Citocinas/efeitos dos fármacos
Formazans
Soluções para Hemodiálise/química
Células Endoteliais da Veia Umbilical Humana/metabolismo
Immunoblotting
Inflamação/metabolismo
Lipopolissacarídeos
NF-kappa B/análise
Óxido Nítrico/análise
Fosforilação
Reação em Cadeia da Polimerase em Tempo Real
Reprodutibilidade dos Testes
Sais de Tetrazólio
Limites: Humanos
Tipo de Publ: Estudo de Avaliação
Responsável: BR1.1 - BIREME


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Ponzoni, Deise
Puricelli, Edela
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Id: biblio-841166
Autor: CORSETTI, Adriana; BAHUSCHEWSKYJ, Claudia; PONZONI, Deise; LANGIE, Renan; SANTOS, Luis Alberto dos; CAMASSOLA, Melissa; NARDI, Nance Beyer; PURICELLI, Edela.
Título: Repair of bone defects using adipose-derived stem cells combined with alpha-tricalcium phosphate and gelatin sponge scaffolds in a rat model
Fonte: J. appl. oral sci;25(1):10-19, Jan.-Feb. 2017. tab, graf.
Idioma: en.
Resumo: Abstract Objectives This study aimed to evaluate the potential of adipose-derived stem cells (ASCs) combined with a modified α-tricalcium phosphate (α-TCP) or gelatin sponge (GS) scaffolds for bone healing in a rat model. Material and Methods Bone defects were surgically created in the femur of adult SHR rats and filled with the scaffolds, empty or combined with ASCs. The results were analyzed by histology and histomorphometry on days seven, 14, 30, and 60. Results Significantly increased bone repair was observed on days seven and 60 in animals treated with α-TCP/ASCs, and on day 14 in the group treated with GS/ASCs, when compared with the groups treated with the biomaterials alone. Intense fibroplasia was observed in the group treated with GS alone, on days 14 and 30. Conclusions Our results showed that the use of ASCs combined with α-TCP or GS scaffolds resulted in increased bone repair. The higher efficacy of the α-TCP scaffold suggests osteoconductive property that results in a biological support to the cells, whereas the GS scaffold functions just as a carrier. These results confirm the potential of ASCs in accelerating bone repair in in vivo experimental rat models. These results suggest a new alternative for treating bone defects.
Descritores: Tecido Adiposo/citologia
Materiais Biocompatíveis/farmacologia
Regeneração Óssea/efeitos dos fármacos
Fosfatos de Cálcio/farmacologia
Esponja de Gelatina Absorvível/farmacologia
Transplante de Células-Tronco/métodos
Tecidos Suporte
-Materiais Biocompatíveis/uso terapêutico
Fosfatos de Cálcio/uso terapêutico
Adesão Celular/efeitos dos fármacos
Proliferação de Células/efeitos dos fármacos
Células Cultivadas
Fêmur/patologia
Fêmur/cirurgia
Fibroblastos/efeitos dos fármacos
Formazans
Esponja de Gelatina Absorvível/uso terapêutico
Modelos Animais
Osteogênese/efeitos dos fármacos
Ratos Endogâmicos SHR
Reprodutibilidade dos Testes
Sais de Tetrazólio
Fatores de Tempo
Resultado do Tratamento
Cicatrização/efeitos dos fármacos
Limites: Animais
Masculino
Tipo de Publ: Estudo de Avaliação
Responsável: BR1.1 - BIREME


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Id: lil-792592
Autor: FERNANDES, Ana Paula; JUNQUEIRA, Marina de Azevedo; MARQUES, Nádia Carolina Teixeira; MACHADO, Maria Aparecida Andrade Moreira; SANTOS, Carlos Ferreira; OLIVEIRA, Thais Marchini; SAKAI, Vivien Thiemy.
Título: Effects of low-level laser therapy on stem cells from human exfoliated deciduous teeth
Fonte: J. appl. oral sci;24(4):332-337, July-Aug. 2016. graf.
Idioma: en.
Resumo: ABSTRACT Low-Level Laser Therapy stimulates the proliferation of a variety of types of cells. However, very little is known about its effect on stem cells from human exfoliated deciduous teeth (SHED). Objective This study aimed to evaluate the influence of different laser therapy energy densities on SHED viability and proliferation. Material and Methods SHED were irradiated according to the groups: I (1.2 J/cm2 - 0.5 mW – 10 s), II (2.5 J/cm2 – 10 mW – 10 s), III (3.7 J/cm2 – 15 mW – 10 s), IV (5.0 J/cm2 – 20 mW – 10 s), V (6.2 J/cm2 – 25 mW – 10 s), and VI (not irradiated – control group). Cell viability was assessed 6 and 24 h after irradiation measuring the mitochondrial activity and using the Crystal Violet assay. Cell proliferation was assessed after 24, 48, and 72 h of irradiation by SRB assay. Results MTT assay demonstrated differences from 6 to 24 hours after irradiation. After 24 h, groups I and IV showed higher absorbance values than those of control group. Crystal Violet assay showed statistically differences in the absorbance rate from 6 to 24 h after irradiation for groups III and VI. At 24 h after irradiation, Group III absorbance rate was greater than that of groups I, II, and IV. Group VI absorbance rate was greater than that of groups I and IV. SRB assay showed that the group I had higher rates than those of groups II, III, V, and VI, at 24 h after irradiation. After 48 h, group I exhibited the greatest cell proliferation rate followed by groups III, V, and VI. After 72 h, group III exhibited the lowest cell proliferation rate than those of groups II, IV, and V. Conclusions The Low-Level Laser Therapy energy densities used in this study did not cause loss of cell viability and stimulated SHED proliferation within the parameters described in this study.
Descritores: Terapia com Luz de Baixa Intensidade/métodos
Células-Tronco/efeitos da radiação
Esfoliação de Dente
Dente Decíduo/citologia
Dente Decíduo/efeitos da radiação
-Análise de Variância
Proliferação de Células/efeitos da radiação
Sobrevivência Celular/efeitos da radiação
Células Cultivadas
Formazans
Doses de Radiação
Reprodutibilidade dos Testes
Rodaminas
Sais de Tetrazólio
Fatores de Tempo
Limites: Humanos
Tipo de Publ: Estudo de Avaliação
Responsável: BR1.1 - BIREME


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Sobral, Maria Angela Pita
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Id: lil-777237
Autor: SOUZA-RODRIGUES, Renata Duarte; FERREIRA, Stella da Silva; D’ALMEIDA-COUTO, Roberta Souza; LACHOWSKI, Karina Monteleone; SOBRAL, Maria Ângela Pita; MARQUES, Márcia Martins.
Título: Choice of toothpaste for the elderly: an in vitro study
Fonte: Braz. oral res. (Online);29(1):1-7, 2015. tab, ilus.
Idioma: en.
Resumo: Hyposalivation and dental root exposure in the elderly are problems that require special oral care. In this context, the characteristics of certain toothpastes are of particular importance. Thus, the aim of this study was to evaluate the cytotoxicity and dentin wear caused by seven different toothpastes. For dentin wear analysis, 40 root dentin specimens were submitted to 20,000 brushing cycles with the different toothpastes and distilled water (control group-CG), using a brushing machine. Dentin surface loss (SL) was measured by contact profilometer. The cytotoxicity of each toothpaste was tested using cultured fibroblasts submitted to a cell-culture-conditioned medium. Fresh medium served as the control. Cell viability was assessed by MTT assay after 24 h of contact with the conditioned media. The data were analyzed statistically by ANOVA, followed by Tukey’s test (p < 0.05). The SL of the CG was minimal and significantly lower than that of the Oral B Pro Health (OBPH) group (p < 0.05). All other groups presented SL in between that of the CG and the Oral B Pro Health OBPH group, except for the Sensodyne (SEN) group, which presented SL similar to that of CG (p = 0.05). The SEN group presented a percentage of viable cells similar to that of CG: between 60-89%. All the other toothpastes showed high cytotoxicity, with cell viability less than 50% of the CG. Considering study limitations, we concluded that only one of the seven tested toothpastes exhibited the most desirable toothpaste characteristics for the worldwide growing elderly population (e.g. low cytotoxicity and low-abrasive potential).
Descritores: Dentina/efeitos dos fármacos
Fibroblastos/efeitos dos fármacos
Desgaste dos Dentes/induzido quimicamente
Cremes Dentais/química
Cremes Dentais/toxicidade
-Análise de Variância
Células Cultivadas
Sobrevivência Celular/efeitos dos fármacos
Dentina/química
Formazans
Teste de Materiais
Propriedades de Superfície/efeitos dos fármacos
Sais de Tetrazólio
Fatores de Tempo
Escovação Dentária
Limites: Idoso
Humanos
Responsável: BR1.1 - BIREME


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Id: lil-773799
Autor: Fernandes, Ana Paula.
Título: Efeitos de TGF-1 em células-tronco pulpares / Effect of Transforming Growth Factor Beta 1(TGF-β 1) in stem cells derived from dental pulp.
Fonte: Bauru; s.n; 2015. 115 p. ilus, tab, graf.
Idioma: pt.
Tese: Apresentada a Universidade de São Paulo. Faculdade de Odontologia de Bauru para obtenção do grau de Doutor.
Resumo: O objetivo deste trabalho foi avaliar, in vitro, os efeitos de diferentes concentrações do fator de crescimento transformador beta 1 (TGF-β1) em células-tronco derivadas da polpa de dentes decíduos esfoliados humanos (SHED), com relação à viabilidade, proliferação, migração e diferenciação celular. As SHED foram mantidas em meio de cultura MEMα + soro fetal bovino (FBS) 10% + penicilina e estreptomicina 1% e tratadas com TGF-β1 na concentração de 1,0; 5,0 e 10,0 ng/mL. Após 1, 3, 5 e 7 dias, foram avaliadas a viabilidade celular pelo método MTT e a proliferação pelo método SRB. Após 24 h de tratamento com TGF-β1, foi realizado um ensaio de migração celular por meio de insertos com poros de 8 μm. Para a avaliação da diferenciação celular de SHED em odontoblastos foram analisados por meio da RT-PCR os marcadores DSPP e DMP-1, após tratamento com TGF-β1 nas diferentes concentrações por 14 dias. Os resultados foram submetidos à ANOVA seguido do teste de Tukey. Em relação à viabilidade celular, as diferentes concentrações de TGF-β1 não tiveram efeito citotóxico sobre SHED. As células tratadas com diferentes concentrações de TGF-β1 apresentaram maiores taxas de proliferação que as do controle negativo (MEMα + 10% de FBS) a partir do 3o dia (p=0,000). Observou-se maiores taxas de migração em direção aos meios contendo TGF-β1, mas sem diferença estatisticamente significativa entre as diferentes concentrações utilizadas, entretanto, houve diferença estatisticamente significativa entre as diferentes concentrações de TGF-β1 com o controle positivo (p=0,000), controle negativo (p=0,000) e entre o controle positivo e negativo (p=0,002). A expressão de DMP-1 foi observada de forma crescente nas doses de 1,0 e 5,0 ng/mL de TGF-β1 ao longo do período (1, 7 e 14 dias) e na dose de 10,0 ng/mL a marcação foi mais intensa desde o primeiro dia do estímulo. Em relação à expressão de DSPP, o grupo tratado com 10,0 ng/mL apresentou marcação após 14 dias de tratamento...

The aim of this study was to evaluate, in vitro, the effect of transforming growth factor beta 1 (TGF-β1) in stem cells derived from the pulp of human exfoliated deciduous teeth (SHED) regarding to cell viability, proliferation, migration and differentiation. SHED were maintained in MEMα culture medium + 10% fetal bovine serum (FBS) + 1% penicillin and streptomycin, and treated with TGF-β1 at the following concentrations of 1.0; 5.0 and 10.0 ng/mL. After 1, 3, 5 and 7 days, cell viability was assessed by MTT assay and proliferation by the SRB method. After 24h of TGF-β1 treatment, cell migration assay was carried out using inserts of 8 μm pore size. To evaluate SHED differentiation into odontoblasts, DMP-1 and DSPP markers were analyzed by RT-PCR, after treatment at different concentrations of TGF-β1 for 14 days. The results were submitted by ANOVA and Tukey test. With respect to cell viability, the different TGF-β1 concentrations did not have cytotoxic effect on SHED. The cells treated by different TGF-β1 concentrations showed higher proliferation rates than those of the negative control (MEMα + 10% FBS) after the third day (p = 0.000). Higher rates of migration towards the media containing TGF-β1 were observed, but there were no statistically significant differences among the concentrations. All different TGF-β1 concentrations showed statistically significant differences with the positive control (p=0.000) and negative control (p=0.000). Statistically significant differences were observed between positive and negative control (p=0.002). DMP-1 expression was observed incrementally at TGF-β1 concentrations of 1.0 and 5.0 ng/mL at 1, 7, and 14 days and the concentration of 10.0 ng/mL was more intense from day one of the stimulus. DSPP expression was more intense after 14 days of treatment with the concentration of 10.0 ng/mL. Thus, this study concluded that different TGF-β1 concentrations stimulated cell proliferation and migration, without...
Descritores: Células-Tronco
Dente Decíduo/citologia
Fator de Crescimento Transformador beta1/farmacologia
-Análise de Variância
Células Cultivadas
Diferenciação Celular
Formazans
Movimento Celular
Proliferação de Células
Reprodutibilidade dos Testes
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Sobrevivência Celular
Fatores de Tempo
Limites: Humanos
Responsável: BR28.1 - Serviço de Biblioteca e Documentação Professor Doutor Antônio Gabriel Atta


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Texto completo SciELO Brasil
Bahia, Maria Terezinha
Texto completo
Id: lil-769833
Autor: Monteiro, Cíntia Júnia; Mota, Suianne Letícia Antunes; Diniz, Lívia de Figueiredo; Bahia, Maria Terezinha; Moraes, Karen CM.
Título: Mir-190b negatively contributes to the Trypanosoma cruzi- infected cell survival by repressing PTEN protein expression
Fonte: Mem. Inst. Oswaldo Cruz;110(8):996-1002, Dec. 2015. graf.
Idioma: en.
Projeto: FAPEMIG; . FAPEMIG; . CNPq.
Resumo: Chagas disease, which is caused by the intracellular protozoanTrypanosoma cruzi, is a serious health problem in Latin America. The heart is one of the major organs affected by this parasitic infection. The pathogenesis of tissue remodelling, particularly regarding cardiomyocyte behaviour after parasite infection, and the molecular mechanisms that occur immediately following parasite entry into host cells are not yet completely understood. Previous studies have reported that the establishment of parasitism is connected to the activation of the phosphatidylinositol-3 kinase (PI3K), which controls important steps in cellular metabolism by regulating the production of the second messenger phosphatidylinositol-3,4,5-trisphosphate. Particularly, the tumour suppressor PTEN is a negative regulator of PI3K signalling. However, mechanistic details of the modulatory activity of PTEN on Chagas disease have not been elucidated. To address this question, H9c2 cells were infected with T. cruzi Berenice 62 strain and the expression of a specific set of microRNAs (miRNAs) were investigated. Our cellular model demonstrated that miRNA-190b is correlated to the decrease of cellular viability rates by negatively modulating PTEN protein expression in T. cruzi-infected cells.
Descritores: Regulação para Baixo
MicroRNAs/fisiologia
Miócitos Cardíacos/parasitologia
Biossíntese de Proteínas
PTEN Fosfo-Hidrolase/metabolismo
Trypanosoma cruzi/metabolismo
-Western Blotting
Linhagem Celular
Sobrevivência Celular
Formazans
Genes Reporter
Miócitos Cardíacos/metabolismo
Fosforilação
PTEN Fosfo-Hidrolase/genética
Reação em Cadeia da Polimerase Via Transcriptase Reversa
RNA Mensageiro/metabolismo
Sais de Tetrazólio
Trypanosoma cruzi/classificação
Limites: Animais
Ratos
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME



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