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1.
Arq. bras. med. vet. zootec. (Online) ; 73(2): 513-516, Mar.-Apr. 2021. graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1248925

RESUMO

A doxorrubicina (dox) é um medicamento antineoplásico que induz cardiotoxicidade por estresse oxidativo. Os flavonoides são antioxidantes extraídos de plantas como Camellia sinensis e Arrabidaea chica (Fridericia chica). Esta pesquisa objetivou avaliar efeitos protetores do extrato de A. chica (AC), comparado ao de C. sinensis (CS), frente ao estresse oxidativo induzido pela dox, no coração. Cardiomiócitos e células neoplásicas MDA-MB 231 foram incubados com AC e CS. Depois, adicionou-se dox e avaliaram-se taxas de viabilidade e morte celular. A citometria de fluxo para o ensaio de iodeto de propídeo (IP) em cardiomiócitos mostrou as seguintes taxas de morte celular: controle 53%; dox 78% (maior que controle, P=0,015); AC_12,5µg/mL + dox 65% (menor que dox, P=0,031); AC_25µg/mL + dox 62% (menor que dox, P=0,028); AC_50µg/mL + dox 63% (menor que dox, P=0,030); CS_12,5µg/mL + dox 71% (menor que dox, P=0,040); CS_25µg/ml + dox 69% (menor que dox, P=0,037); CS_50µg/mL + dox 74% (menor que dox, P=0,044). Resultados das células MDA-MB 231 mostraram que nenhum extrato interferiu na atividade antitumoral da dox. Os dados de IP foram corroborados pelos de MTT. Este estudo reporta promissora utilização de A. chica na prevenção da cardiotoxicidade induzida pela dox.(AU)


Assuntos
Animais , Ratos , Extratos Vegetais/uso terapêutico , Doxorrubicina , Bignoniaceae/química , Cardiotoxicidade/terapia , Cardiotoxicidade/veterinária , Plantas Medicinais , Flavonoides/uso terapêutico
2.
Arq. bras. med. vet. zootec. (Online) ; 71(4): 1121-1130, jul.-ago. 2019. graf, ilus
Artigo em Português | VETINDEX | ID: vti-25199

RESUMO

O objetivo do estudo foi avaliar o efeito da matriz porosa do biovidro 60S (BV60S) associada a células osteoprogenitoras (CO) alógenas no tratamento de defeitos ósseos críticos de cães. Foram utilizados 20 cães, machos, sem raça definida, com dois anos de idade e massa corporal média de 25kg. Com os cães sob anestesia geral, foram criados defeitos ósseos críticos no terço médio dos ossos rádios. Procedeu-se à fixação óssea com uma placa em ponte, e os defeitos foram tratados de acordo com cada grupo experimental. Constituíram-se três grupos experimentais, em que os defeitos ósseos foram preenchidos com: BV60S associado a CO alógenas (grupo BV60S+CO), osso autógeno (grupo C+), ou não preenchidos (grupo C-). A regeneração óssea foi avaliada por meio de exames radiográficos, densitométricos e histomorfométricos ao longo de 90 dias. Os grupos C- e BV60S+CO mostraram preenchimento ósseo parcial do defeito de, no máximo, 56,68% e 35,23%, respectivamente, sem a formação de ponte óssea entre as extremidades, e o controle positivo (C+) mostrou regeneração óssea completa. Conclui-se que a matriz porosa do BV60S associada às células osteoprogenitoras não é eficiente no tratamento de defeitos ósseos críticos em rádios de cães.(AU)


The objective of this study was to evaluate the effect of the porous matrix of bioglass 60S (BV60S) associated with allogenic osteoprogenitor cells (CO) in the treatment of critical bone defects of dogs. 20 male mongrel dogs at two years old and mean weight of 25kg were used. Dogs were anesthetized and critical bone defects were created in the middle third of the radios bones. With dogs under general anesthesia, critical bone defects were created in the middle third of bone radios. Bone fixation was done with a bridge plate and defects treated according to each experimental group. Three experimental groups were formed according to the treatment. The defects filled with BV60S associated with allogenic CO (Group-BV60S+CO), autogenous bone (Group-C+) or unfilled (Group-C-). Bone regeneration was evaluated by radiography, bone densitometry and histomorphometry over 90 days. The BV60S+CO and C- groups showed partial bone filling of the defect of at most 56.68% and 35.23%, respectively. No bone bridge formation was observed between the extremities in the BV60S+CO and C- groups. Positive control showed complete bone regeneration at 90 days. It was concluded that the porous matrix of BV60S associated with osteoprogenitor cells was not effective in the treatment of critical bone defects in the radius of dogs.(AU)


Assuntos
Animais , Cães , Rádio (Anatomia)/lesões , Materiais Biocompatíveis/uso terapêutico , Regeneração Óssea , Terapia Baseada em Transplante de Células e Tecidos/métodos , Terapia Baseada em Transplante de Células e Tecidos/veterinária
3.
Arq. bras. med. vet. zootec. (Online) ; 71(4): 1121-1130, jul.-ago. 2019. graf, ilus
Artigo em Português | LILACS, VETINDEX | ID: biblio-1038624

RESUMO

O objetivo do estudo foi avaliar o efeito da matriz porosa do biovidro 60S (BV60S) associada a células osteoprogenitoras (CO) alógenas no tratamento de defeitos ósseos críticos de cães. Foram utilizados 20 cães, machos, sem raça definida, com dois anos de idade e massa corporal média de 25kg. Com os cães sob anestesia geral, foram criados defeitos ósseos críticos no terço médio dos ossos rádios. Procedeu-se à fixação óssea com uma placa em ponte, e os defeitos foram tratados de acordo com cada grupo experimental. Constituíram-se três grupos experimentais, em que os defeitos ósseos foram preenchidos com: BV60S associado a CO alógenas (grupo BV60S+CO), osso autógeno (grupo C+), ou não preenchidos (grupo C-). A regeneração óssea foi avaliada por meio de exames radiográficos, densitométricos e histomorfométricos ao longo de 90 dias. Os grupos C- e BV60S+CO mostraram preenchimento ósseo parcial do defeito de, no máximo, 56,68% e 35,23%, respectivamente, sem a formação de ponte óssea entre as extremidades, e o controle positivo (C+) mostrou regeneração óssea completa. Conclui-se que a matriz porosa do BV60S associada às células osteoprogenitoras não é eficiente no tratamento de defeitos ósseos críticos em rádios de cães.(AU)


The objective of this study was to evaluate the effect of the porous matrix of bioglass 60S (BV60S) associated with allogenic osteoprogenitor cells (CO) in the treatment of critical bone defects of dogs. 20 male mongrel dogs at two years old and mean weight of 25kg were used. Dogs were anesthetized and critical bone defects were created in the middle third of the radios bones. With dogs under general anesthesia, critical bone defects were created in the middle third of bone radios. Bone fixation was done with a bridge plate and defects treated according to each experimental group. Three experimental groups were formed according to the treatment. The defects filled with BV60S associated with allogenic CO (Group-BV60S+CO), autogenous bone (Group-C+) or unfilled (Group-C-). Bone regeneration was evaluated by radiography, bone densitometry and histomorphometry over 90 days. The BV60S+CO and C- groups showed partial bone filling of the defect of at most 56.68% and 35.23%, respectively. No bone bridge formation was observed between the extremities in the BV60S+CO and C- groups. Positive control showed complete bone regeneration at 90 days. It was concluded that the porous matrix of BV60S associated with osteoprogenitor cells was not effective in the treatment of critical bone defects in the radius of dogs.(AU)


Assuntos
Animais , Cães , Rádio (Anatomia)/lesões , Materiais Biocompatíveis/uso terapêutico , Regeneração Óssea , Terapia Baseada em Transplante de Células e Tecidos/métodos , Terapia Baseada em Transplante de Células e Tecidos/veterinária
4.
Arq. bras. med. vet. zootec. (Online) ; 70(3): 857-872, Maio-Jun. 2018. ilus, tab, graf
Artigo em Português | VETINDEX | ID: vti-735077

RESUMO

Com o objetivo de estudar o efeito da condroitinase associada às células-tronco mesenquimais na lesão aguda da medula espinhal, utilizaram-se 50 ratos Lewis, distribuídos igualmente nos grupos: controle negativo (CN), tratamento com placebo (PLA), condroitinase (CDN), células-tronco mesenquimais (CTM) e condroitinase mais células-tronco mesenquimais (CDN+CTM). Todos os animais tiveram a medula espinhal exposta por laminectomia, e os grupos PLA, CDT, CTM e CDT+CTM sofreram também trauma medular compressivo. Após sete dias, procedeu-se à reexposição da medula espinhal, quando os grupos PLA e CTM receberam 4µL de líquido cefalorraquidiano artificial via intralesional, e os grupos CDT e CDT+CTM receberam o mesmo líquido contendo 2,2U de condroitinase. Após 14 dias da cirurgia inicial, todos os animais receberam 0,2mL de PBS via endovenosa, contudo, nos grupos CTM e CDT+CTM, esse líquido continha 1x106 CTM. Avaliou-se a capacidade motora até o 28o dia pós-trauma e, posteriormente, as medulas espinhais foram analisadas por RT-PCR, para quantificação da expressão gênica para BDNF, NT-3, VEGF, KDR e PECAM-1, e por imunoistoquímica, para detecção das células-tronco GFP injetadas (anti-GFP), quantificação dos neurônios (anti-NeuN) e da GFAP e vimentina, para avaliação da cicatriz glial. As análises estatísticas foram realizadas com o auxílio do Prism 5 for Windows, com o nível de significância de 5%. Não houve diferença entre os grupos quanto à capacidade motora. O grupo CDT+CTM apresentou maior imunoexpressão de neurônios viáveis do que o placebo. No CTM, houve maior expressão dos fatores neurotróficos BDNF e VEGF. E no CDT, houve menor imunoexpressão de vimentina. Concluiu-se que a associação CDT+CTM favorece a viabilidade neuronal após o trauma, que o tratamento com CTM promove aumento na expressão dos fatores tróficos BDNF e VEGF e que o tratamento com condroitinase é efetivo na redução da cicatriz glial.(AU)


The aim of this work was to study the effect of chondroitinase associated with mesenchymal stem cells in acute spinal cord injury. Therefore, 50 Lewis rats were distributed in the following groups: negative control (NC), treatment with placebo (PLA), chondroitinase (CDT), mesenchymal stem cells (MSC), and chondroitinase associated with mesenchymal stem cells (CDT + MSC). All animals had their spinal cord exposed by laminectomy, and the groups named PLA, CDT, MSC and CDT + MSC also suffered compressive spinal cord trauma. After seven days, the spinal cord was re-exposed, when the PLA and MSCs groups received 4uL of artificial cerebrospinal fluid through the lesion, and the CDT group and CDT + MSC received the same fluid containing 2,2U of chondroitinase. 14 days after the first surgery, all animals received 0.2ml of PBS intravenously; however, the MSC and CDT + MSC groups received the same liquid also containing 1x106 MSCs. The motor skills were evaluated up to 28 days post-injury and, subsequently, the spinal cords were analyzed by RT-PCR for BDNF, NT-3, VEGF, PECAM-1 and KDR gene expression quantification, immunohistochemistry to detect injected stem cells GFP (anti-GFP), to quantify neurons (anti-NeuN), GFAP and detect vimentin in order to evaluate the glial scar. Statistical analyzes were performed by Prism 5 for Windows using a 5% level of significance. There was no difference between groups with regarding motor capacity. The CDT + MSC group showed increased immunoreactivity of viable neurons than placebo. In MSC, there was a greater expression of neurotrophic factors BDNF and VEGF. Also, there was less vimentin immunostaining in group CDT. It was concluded that CDT + MSC association promotes neuronal viability after trauma, in which treatment with MSC promotes increased expression of BDNF and VEGF trophic factors, and also that treatment with chondroitinase is effective in reducing the glial scar.(AU)


Assuntos
Animais , Ratos , Condroitina ABC Liase , Ratos/anatomia & histologia , Ratos/lesões , Células-Tronco Mesenquimais/enzimologia
5.
Arq. bras. med. vet. zootec. (Online) ; 70(3): 857-872, maio-jun. 2018. ilus, tab, graf
Artigo em Português | LILACS, VETINDEX | ID: biblio-911635

RESUMO

Com o objetivo de estudar o efeito da condroitinase associada às células-tronco mesenquimais na lesão aguda da medula espinhal, utilizaram-se 50 ratos Lewis, distribuídos igualmente nos grupos: controle negativo (CN), tratamento com placebo (PLA), condroitinase (CDN), células-tronco mesenquimais (CTM) e condroitinase mais células-tronco mesenquimais (CDN+CTM). Todos os animais tiveram a medula espinhal exposta por laminectomia, e os grupos PLA, CDT, CTM e CDT+CTM sofreram também trauma medular compressivo. Após sete dias, procedeu-se à reexposição da medula espinhal, quando os grupos PLA e CTM receberam 4µL de líquido cefalorraquidiano artificial via intralesional, e os grupos CDT e CDT+CTM receberam o mesmo líquido contendo 2,2U de condroitinase. Após 14 dias da cirurgia inicial, todos os animais receberam 0,2mL de PBS via endovenosa, contudo, nos grupos CTM e CDT+CTM, esse líquido continha 1x106 CTM. Avaliou-se a capacidade motora até o 28o dia pós-trauma e, posteriormente, as medulas espinhais foram analisadas por RT-PCR, para quantificação da expressão gênica para BDNF, NT-3, VEGF, KDR e PECAM-1, e por imunoistoquímica, para detecção das células-tronco GFP injetadas (anti-GFP), quantificação dos neurônios (anti-NeuN) e da GFAP e vimentina, para avaliação da cicatriz glial. As análises estatísticas foram realizadas com o auxílio do Prism 5 for Windows, com o nível de significância de 5%. Não houve diferença entre os grupos quanto à capacidade motora. O grupo CDT+CTM apresentou maior imunoexpressão de neurônios viáveis do que o placebo. No CTM, houve maior expressão dos fatores neurotróficos BDNF e VEGF. E no CDT, houve menor imunoexpressão de vimentina. Concluiu-se que a associação CDT+CTM favorece a viabilidade neuronal após o trauma, que o tratamento com CTM promove aumento na expressão dos fatores tróficos BDNF e VEGF e que o tratamento com condroitinase é efetivo na redução da cicatriz glial.(AU)


The aim of this work was to study the effect of chondroitinase associated with mesenchymal stem cells in acute spinal cord injury. Therefore, 50 Lewis rats were distributed in the following groups: negative control (NC), treatment with placebo (PLA), chondroitinase (CDT), mesenchymal stem cells (MSC), and chondroitinase associated with mesenchymal stem cells (CDT + MSC). All animals had their spinal cord exposed by laminectomy, and the groups named PLA, CDT, MSC and CDT + MSC also suffered compressive spinal cord trauma. After seven days, the spinal cord was re-exposed, when the PLA and MSCs groups received 4uL of artificial cerebrospinal fluid through the lesion, and the CDT group and CDT + MSC received the same fluid containing 2,2U of chondroitinase. 14 days after the first surgery, all animals received 0.2ml of PBS intravenously; however, the MSC and CDT + MSC groups received the same liquid also containing 1x106 MSCs. The motor skills were evaluated up to 28 days post-injury and, subsequently, the spinal cords were analyzed by RT-PCR for BDNF, NT-3, VEGF, PECAM-1 and KDR gene expression quantification, immunohistochemistry to detect injected stem cells GFP (anti-GFP), to quantify neurons (anti-NeuN), GFAP and detect vimentin in order to evaluate the glial scar. Statistical analyzes were performed by Prism 5 for Windows using a 5% level of significance. There was no difference between groups with regarding motor capacity. The CDT + MSC group showed increased immunoreactivity of viable neurons than placebo. In MSC, there was a greater expression of neurotrophic factors BDNF and VEGF. Also, there was less vimentin immunostaining in group CDT. It was concluded that CDT + MSC association promotes neuronal viability after trauma, in which treatment with MSC promotes increased expression of BDNF and VEGF trophic factors, and also that treatment with chondroitinase is effective in reducing the glial scar.(AU)


Assuntos
Animais , Ratos , Condroitina ABC Liase , Ratos/anatomia & histologia , Ratos/lesões , Células-Tronco Mesenquimais/enzimologia
6.
Arq. bras. med. vet. zootec. (Online) ; 69(6): 1573-1580, Nov.-Dez. 2017. tab, graf
Artigo em Inglês | VETINDEX | ID: vti-735000

RESUMO

The objective was to evaluate the in vitro effect of prolactin in osteogenic potential of adipose tissue-derived mesenchymal stem cells (ADSCs) in female rats. ADSCs were cultured in osteogenic medium with and without the addition of prolactin and distributed into three groups: 1) ADSCs (control), 2) ADSCs with addition of 100ng/mL of prolactin and 3) ADSCs with addition of 300ng/mL of prolactin. At 21 days of differentiation, the tests of MTT conversion into formazan crystals, percentage of mineralized nodules and cells per field and quantification of genic transcript for alkaline phosphatase, osteopontin, osteocalcin, bone sialoprotein, BMP-2 and collagen I by real-time RT-PCR were made. The addition of prolactin reduced the conversion of MTT in group 3 and increased the percentage of cells per field in the groups 2 and 3, however without significantly increasing the percentage of mineralized nodules and the expression of alkaline phosphatase, osteopontin, osteocalcin, bone sialoprotein, BMP-2 and collagen I. In conclusion, the addition of prolactin in concentrations of 100ng/mL and 300ng/mL does not change the osteogenic differentiation to the ADSCs of female rats despite increase in the cellularity of the culture.(AU)


O objetivo do presente trabalho foi avaliar o efeito in vitro da prolactina sobre o potencial osteogênico de células-tronco mesenquimais do tecido adiposo (CTM-TA) em ratas. CTM-TA foram cultivadas em meio osteogênico com e sem adição de prolactina e distribuídas em três grupos: 1) CTM-TA (controle), 2) CM-TA com adição de 100ng/mL de prolactina e 3) CTM-TA com adição de 300ng/mL de prolactina. Aos 21 dias de diferenciação, foram realizados os testes de conversão do MTT em cristais de formazan, porcentagem de nódulos mineralizados e células por campo e quantificação dos transcritos gênicos para fosfatase alcalina, osteopontina, osteocalcina, sialoproteína óssea, BMP-2 e colágeno I. A adição de prolactina reduziu a conversão do MTT no grupo 3 e aumentou a porcentagem de células por campo nos grupos 2 e 3, sem alterar significativamente a porcentagem de nódulos mineralizados e a expressão de fosfatase alcalina, osteopontina, osteocalcina, sialoproteína óssea, BMP-2 e colágeno I. Conclui-se que a adição de prolactina nas concentrações de 100ng/mL e 300ng/mL não altera a diferenciação osteogênica das CTM-TA de ratas, apesar do aumento de celularidade da cultura.(AU)


Assuntos
Animais , Feminino , Ratos , Tecido Adiposo , Osteogênese , Prolactina/análise , Células-Tronco , Osteoblastos
7.
Arq. bras. med. vet. zootec. (Online) ; 69(6): 1573-1580, nov.-dez. 2017. tab, graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-910772

RESUMO

The objective was to evaluate the in vitro effect of prolactin in osteogenic potential of adipose tissue-derived mesenchymal stem cells (ADSCs) in female rats. ADSCs were cultured in osteogenic medium with and without the addition of prolactin and distributed into three groups: 1) ADSCs (control), 2) ADSCs with addition of 100ng/mL of prolactin and 3) ADSCs with addition of 300ng/mL of prolactin. At 21 days of differentiation, the tests of MTT conversion into formazan crystals, percentage of mineralized nodules and cells per field and quantification of genic transcript for alkaline phosphatase, osteopontin, osteocalcin, bone sialoprotein, BMP-2 and collagen I by real-time RT-PCR were made. The addition of prolactin reduced the conversion of MTT in group 3 and increased the percentage of cells per field in the groups 2 and 3, however without significantly increasing the percentage of mineralized nodules and the expression of alkaline phosphatase, osteopontin, osteocalcin, bone sialoprotein, BMP-2 and collagen I. In conclusion, the addition of prolactin in concentrations of 100ng/mL and 300ng/mL does not change the osteogenic differentiation to the ADSCs of female rats despite increase in the cellularity of the culture.(AU)


O objetivo do presente trabalho foi avaliar o efeito in vitro da prolactina sobre o potencial osteogênico de células-tronco mesenquimais do tecido adiposo (CTM-TA) em ratas. CTM-TA foram cultivadas em meio osteogênico com e sem adição de prolactina e distribuídas em três grupos: 1) CTM-TA (controle), 2) CM-TA com adição de 100ng/mL de prolactina e 3) CTM-TA com adição de 300ng/mL de prolactina. Aos 21 dias de diferenciação, foram realizados os testes de conversão do MTT em cristais de formazan, porcentagem de nódulos mineralizados e células por campo e quantificação dos transcritos gênicos para fosfatase alcalina, osteopontina, osteocalcina, sialoproteína óssea, BMP-2 e colágeno I. A adição de prolactina reduziu a conversão do MTT no grupo 3 e aumentou a porcentagem de células por campo nos grupos 2 e 3, sem alterar significativamente a porcentagem de nódulos mineralizados e a expressão de fosfatase alcalina, osteopontina, osteocalcina, sialoproteína óssea, BMP-2 e colágeno I. Conclui-se que a adição de prolactina nas concentrações de 100ng/mL e 300ng/mL não altera a diferenciação osteogênica das CTM-TA de ratas, apesar do aumento de celularidade da cultura.(AU)


Assuntos
Animais , Feminino , Ratos , Tecido Adiposo , Osteogênese , Prolactina/análise , Células-Tronco , Osteoblastos
8.
Genet Mol Res ; 16(1)2017 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-28340277

RESUMO

This study aimed to evaluate the immunomodulatory and neuroprotective effects of allogeneic and cryopreserved mesenchymal stem cells (MSCs) on spinal cord injury. A total of 120 rats were distributed into the following groups: negative control (NC) - without injury, positive control (PC) - with injury without treatment, and group treated with MSC (GMSC) - with injury and treated. Motor function was evaluated by the BBB test at 24, 48, and 72 h and at 8 and 21 postoperative days. Spinal cords were evaluated by histopathology and immunohistochemistry to determine the expression of CD68, NeuN, and GFAP. IL-10, TNF-α, IL-1ß, TGF-ß, BDNF, GDNF, and VEGF expression was quantified by RT-PCR. The GMSC presented higher scores for motor function at 72 h and 8 and 21 days after injury, lower expression of CD68 at 8 days, and lower expression of GFAP at 21 days compared to the PC. In addition, higher expression of NeuN and lower degeneration of the white matter occurred at 21 days. The GMSC also showed higher expression of IL-10 24 h after injury, GDNF at 48 h and 8 days, and VEGF at 21 days. Moreover, lower expression of TNF-α was observed at 8 and 21 days and TGF-ß at 24 h and 21 days. There were no differences in the expression of IL-1ß and BDNF between the GMSC and PC. Thus, cryopreserved MSCs promote immunomodulatory and neuroprotective effects in rats with spinal cord injury by increasing IL-10, GDNF, and VEGF expression and reducing TNF-α and TGF-ß expression.


Assuntos
Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/fisiologia , Medicina Regenerativa/métodos , Traumatismos da Medula Espinal/terapia , Animais , Criopreservação/métodos , Modelos Animais de Doenças , Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Interleucina-10/metabolismo , Masculino , Células-Tronco Mesenquimais/imunologia , Células-Tronco Mesenquimais/metabolismo , Fármacos Neuroprotetores/imunologia , Ratos , Ratos Endogâmicos Lew , Traumatismos da Medula Espinal/imunologia , Fator de Crescimento Transformador beta/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo
9.
Artigo em Inglês | VETINDEX | ID: vti-18682

RESUMO

ABSTRACT The objective was to evaluate the in vitro effect of prolactin in osteogenic potential of adipose tissue-derived mesenchymal stem cells (ADSCs) in female rats. ADSCs were cultured in osteogenic medium with and without the addition of prolactin and distributed into three groups: 1) ADSCs (control), 2) ADSCs with addition of 100ng/mL of prolactin and 3) ADSCs with addition of 300ng/mL of prolactin. At 21 days of differentiation, the tests of MTT conversion into formazan crystals, percentage of mineralized nodules and cells per field and quantification of genic transcript for alkaline phosphatase, osteopontin, osteocalcin, bone sialoprotein, BMP-2 and collagen I by real-time RT-PCR were made. The addition of prolactin reduced the conversion of MTT in group 3 and increased the percentage of cells per field in the groups 2 and 3, however without significantly increasing the percentage of mineralized nodules and the expression of alkaline phosphatase, osteopontin, osteocalcin, bone sialoprotein, BMP-2 and collagen I. In conclusion, the addition of prolactin in concentrations of 100ng/mL and 300ng/mL does not change the osteogenic differentiation to the ADSCs of female rats despite increase in the cellularity of the culture.


RESUMO O objetivo do presente trabalho foi avaliar o efeito in vitro da prolactina sobre o potencial osteogênico de células-tronco mesenquimais do tecido adiposo (CTM-TA) em ratas. CTM-TA foram cultivadas em meio osteogênico com e sem adição de prolactina e distribuídas em três grupos: 1) CTM-TA (controle), 2) CM-TA com adição de 100ng/mL de prolactina e 3) CTM-TA com adição de 300ng/mL de prolactina. Aos 21 dias de diferenciação, foram realizados os testes de conversão do MTT em cristais de formazan, porcentagem de nódulos mineralizados e células por campo e quantificação dos transcritos gênicos para fosfatase alcalina, osteopontina, osteocalcina, sialoproteína óssea, BMP-2 e colágeno I. A adição de prolactina reduziu a conversão do MTT no grupo 3 e aumentou a porcentagem de células por campo nos grupos 2 e 3, sem alterar significativamente a porcentagem de nódulos mineralizados e a expressão de fosfatase alcalina, osteopontina, osteocalcina, sialoproteína óssea, BMP-2 e colágeno I. Conclui-se que a adição de prolactina nas concentrações de 100ng/mL e 300ng/mL não altera a diferenciação osteogênica das CTM-TA de ratas, apesar do aumento de celularidade da cultura.

10.
Arq. bras. med. vet. zootec. (Online) ; 67(4): 969-978, jul.-ago. 2015. tab, ilus
Artigo em Português | VETINDEX | ID: vti-324240

RESUMO

O objetivo do presente trabalho foi avaliar o produto iônico do biovidro 60S (BV60S) na diferenciação osteogênica de células-tronco mesenquimais de origem adiposa (CTM-AD) de cães. As CTM-AD foram diferenciadas sem OSTe com o produto iônico (PI OST) por sete, 14 e 21 dias. Avaliou-se o MTT, a fosfatase alcalina (FA), o colágeno, mineralização e as expressões de osterix (OSX), sialoproteína óssea (BSP), osteonectina (ON) e osteocalcina (OC). O grupo PI OSTmostrou menor conversão de MTT aos sete dias e maior conversão aos 21 dias. A atividade de FA foi maior no grupo OST, aos 14 e 21 dias. A síntese de colágeno foi maior no grupo OST aos sete e 21 dias. Verificou-se maior área mineralizada no grupo PI OSTem todos os tempos. Não houve diferenças nas expressões de OSX e OC entre os grupos. Observou-se maior expressão de BSP no grupo PI OST, aos 14 e 21 dias. A expressão de ON foi maior no grupo OST aos 14 dias. Concluiu-se que o produto iônico do BV60S favorece a osteogênese in vitro de CTM-AD de cães.(AU)


The aim was to evaluate the ionic product of 60S bioglass (BV60S) in osteogenic differentiation of mesenchymal stem cells from adipose tissue (ADMSCs) in dogs. ADMSCs were differentiated without the ionic product (OST) and with the ionic product (PI-OST) for 7, 14 and 21 days. We evaluated the MTT, alkaline phosphatase (ALP), collagen mineralization and expressions of osterix (OSX), bone sialoprotein (BSP), osteonectin (ON) and osteocalcin (OC). The PI-OST group had a lower MTT conversion to 7days and higher conversion at 21 days. The ALP activity was higher in the OST group at 14 and 21 days. Collagen synthesis was higher in the OST group at 7 and 21 days. A higher mineralized area in the PI-OST group was observed at all times. There were no differences in expressions of OSX and OC between groups. We observed increased expression of BSP in the PI-OST group at 14 and 21 days. The expression of ON was higher in the OST group at 14 days. It was concluded that the ionic product of BV60S promotes in vitro osteogenesis of MSC-AD from dogs.(AU)


Assuntos
Animais , Cães , Células-Tronco , Osteogênese , Materiais Biocompatíveis , Canais Iônicos , Regeneração Óssea
11.
Arq. bras. med. vet. zootec ; 67(4): 969-978, July-Aug. 2015. tab, ilus
Artigo em Português | LILACS | ID: lil-759238

RESUMO

O objetivo do presente trabalho foi avaliar o produto iônico do biovidro 60S (BV60S) na diferenciação osteogênica de células-tronco mesenquimais de origem adiposa (CTM-AD) de cães. As CTM-AD foram diferenciadas sem OSTe com o produto iônico (PI OST) por sete, 14 e 21 dias. Avaliou-se o MTT, a fosfatase alcalina (FA), o colágeno, mineralização e as expressões de osterix (OSX), sialoproteína óssea (BSP), osteonectina (ON) e osteocalcina (OC). O grupo PI OSTmostrou menor conversão de MTT aos sete dias e maior conversão aos 21 dias. A atividade de FA foi maior no grupo OST, aos 14 e 21 dias. A síntese de colágeno foi maior no grupo OST aos sete e 21 dias. Verificou-se maior área mineralizada no grupo PI OSTem todos os tempos. Não houve diferenças nas expressões de OSX e OC entre os grupos. Observou-se maior expressão de BSP no grupo PI OST, aos 14 e 21 dias. A expressão de ON foi maior no grupo OST aos 14 dias. Concluiu-se que o produto iônico do BV60S favorece a osteogênese in vitro de CTM-AD de cães.


The aim was to evaluate the ionic product of 60S bioglass (BV60S) in osteogenic differentiation of mesenchymal stem cells from adipose tissue (ADMSCs) in dogs. ADMSCs were differentiated without the ionic product (OST) and with the ionic product (PI-OST) for 7, 14 and 21 days. We evaluated the MTT, alkaline phosphatase (ALP), collagen mineralization and expressions of osterix (OSX), bone sialoprotein (BSP), osteonectin (ON) and osteocalcin (OC). The PI-OST group had a lower MTT conversion to 7days and higher conversion at 21 days. The ALP activity was higher in the OST group at 14 and 21 days. Collagen synthesis was higher in the OST group at 7 and 21 days. A higher mineralized area in the PI-OST group was observed at all times. There were no differences in expressions of OSX and OC between groups. We observed increased expression of BSP in the PI-OST group at 14 and 21 days. The expression of ON was higher in the OST group at 14 days. It was concluded that the ionic product of BV60S promotes in vitro osteogenesis of MSC-AD from dogs.


Assuntos
Animais , Cães , Materiais Biocompatíveis , Osteogênese , Células-Tronco , Regeneração Óssea , Canais Iônicos
12.
Acta Crystallogr F Struct Biol Commun ; 70(Pt 6): 803-7, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24915098

RESUMO

Schistosomiasis is an inflammatory chronic disease that represents a major health problem in tropical and subtropical countries. The drug of choice for treatment, praziquantel, is effective in killing adult worms but fails to kill immature forms and prevent reinfection. One prominent antigen candidate for an anti-schistosomiasis vaccine is the protein Sm21.7 (184 amino-acid residues) from Schistosoma mansoni, a tegumental protein capable of reducing the worm burden in a murine immunization model. In the present work, the Sm21.7 gene was cloned and expressed in Escherichia coli and the full-length protein was purified to homogeneity. Crystals of recombinant Sm21.7 suitable for X-ray diffraction were obtained using PEG monomethyl ether 2000 as a precipitant. X-ray diffraction images of a native crystal (at 2.05 Šresolution) and a quick-cryosoaked NaI derivative (at 1.95 Šresolution) were collected on the W01B-MX2 beamline at the Laboratório Nacional de Luz Síncrotron (LNLS, Brazilian Synchrotron Light Laboratory/MCT). Both crystals belonged to the hexagonal space group P6122, with similar unit-cell parameters a=b=108.5, c=55.8 Å. SIRAS-derived phases were used to generate the first electron-density map, from which a partial three-dimensional model of Sm21.7 (from Gln89 to Asn184) was automatically constructed. Anaysis of dissolved crystals by SDS-PAGE confirmed that the protein was cleaved in the crystallization drop and only the Sm21.7 C-terminal domain was crystallized. The structure of the Sm21.7 C-terminal domain will help in the localization of the epitopes responsible for its protective immune responses, constituting important progress in the development of an anti-schistosomiasis vaccine.


Assuntos
Antígenos de Protozoários/imunologia , Schistosoma mansoni/imunologia , Animais , Sequência de Bases , Cristalografia por Raios X , Primers do DNA
13.
J Biomed Mater Res A ; 102(3): 818-27, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23625853

RESUMO

Tissue engineering is a multidisciplinary science that combines a structural scaffold and cells to form a construct able to promote regeneration of injured tissue. Bioactive glass foam produced by sol-gel is an osteoinductive material with a network of interconnected macropores necessary for cell colonization. The use of human adipose-derived stem cell (hASC) presents advantages as the potential for a large number of cells, rapid expansion in vitro and the capability of differentiating into osteoblasts. The use of a bioreactor in three-dimensional cell culture enables greater efficiency for cell nutrition and application of mechanical forces, important modulators of bone physiology. The hASC seeded in a bioactive glass scaffold and cultured in osteogenic Leibovitz L-15 medium in a bioreactor with a flow rate of 0.1 mL min(-1) demonstrated a significant increase in cell proliferation and viability and alkaline phosphatase (ALP) activity peak after 14 days. The immunofluorescence assay revealed an expression of osteopontin, osteocalcin and type I collagen from 7 to 21 days after culture. The cells changed from a spindle shape to a cuboidal morphology characteristic of osteoblasts. The polymerase chain reaction assay confirmed that osteopontin, osteocalcin, and ALP genes were expressed. These results indicate that hASCs differentiated into an osteogenic phenotype when cultured in bioactive glass scaffold, osteogenic Leibovitz L-15 medium and a perfusion bioreactor. Therefore, these results highlight the synergism between a bioactive glass scaffold and the effect of perfusion on cells and indicate the differentiation into an osteogenic phenotype.


Assuntos
Tecido Adiposo/citologia , Materiais Biocompatíveis/química , Reatores Biológicos , Vidro/química , Osteogênese , Células-Tronco/citologia , Engenharia Tecidual/instrumentação , Diferenciação Celular , Proliferação de Células , Sobrevivência Celular , Células Cultivadas , Feminino , Humanos , Osteoblastos/citologia
14.
Neuroscience ; 250: 715-32, 2013 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-23896572

RESUMO

Compression of spinal roots is an important medical problem, which may arise from intervertebral disc herniation, tumor growth or as a result of high energy accidents. Differently from avulsion, root crushing maintains the central/peripheral nervous system (CNS/PNS) connection, although the axons are axotomized and motoneurons degenerate. Such neuronal death may decrease and delay motor function recovery. In the present study we have investigated the neuroprotective effects of mesenchymal stem cell (MSC) therapy following such proximal lesions. Motor recovery and synaptic stabilization were analyzed by the use of morphological and functional approaches. For that, crushing the ventral roots at L4, L5 and L6 was unilaterally performed in Lewis rats. Four weeks after injury, an increased motoneuron survival was observed in the MSC-treated group, coupled with a smaller decrease of inputs at the motoneuron surface and nearby neuropil, seen by synaptophysin and synapsin immunolabeling and decreased astrogliosis, seen by GFAP immunolabeling. In this sense, MSC-treated group displayed a significant preservation of GABAergic terminals, indicating a possible neuroprotection to glutamate excitotoxicity. Motor function recovery was acutely improved in MSC-treated group as compared to Dulbeco's modified eagle medium (DMEM)-treated. Overall, we provide evidence that ventral root crushing (VRC), although milder than avulsion, results in significant loss of motoneurons (~51%) that can be reduced by MSC administration within the spinal cord. Such treatment also improves the number of synapses immunoreactive against molecules present in inhibitory inputs. Also, an increased number of regenerated axons was obtained in the MSC-treated group, in comparison to the DMEM-treated control. Overall, MSC therapy acutely improved limb strength and gait coordination, indicating a possible clinical application of such treatment following proximal lesions at the CNS/PNS interface.


Assuntos
Axotomia , Transplante de Células-Tronco Mesenquimais , Neurônios Motores/fisiologia , Regeneração Nervosa/fisiologia , Medula Espinal/citologia , Raízes Nervosas Espinhais/fisiologia , Sinapses/fisiologia , Animais , Sobrevivência Celular , Células Cultivadas , Feminino , Citometria de Fluxo , Marcha/fisiologia , Imuno-Histoquímica , Força Muscular , Compressão Nervosa , Neuroglia/fisiologia , Ratos , Ratos Endogâmicos Lew , Recuperação de Função Fisiológica , Nervo Isquiático/citologia , Nervo Isquiático/fisiologia
15.
Mycopathologia ; 174(2): 93-105, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22391822

RESUMO

The conventional treatment for the most prevalent mycosis in Latin America, paracoccidioidomycosis (PCM), involves long periods of therapy that results in side effects and a high frequency of relapses. The search for a new, alternative treatment is necessary. Pb40 is an antigenic protein from P. brasiliensis fraction F0. This fraction has already been shown to have significant protective activity when used as a PCM vaccine in experimental models. The complete cDNA sequence corresponding to Pb40 was cloned into a pET-21a plasmid, expressed in E. coli with a his-tag and purified by affinity chromatography. The predicted protein sequence exhibited nearly 100% homology to a fragment of the hypothetical EF-hand domain containing protein of P. brasiliensis. Immunization with this recombinant protein was used together with chemotherapy in an attempt to improve PCM treatment. The combined drug/rPb40 treatment exhibited long-lasting control of PCM in the liver and spleen and largely preserved the tissue structures of these organs. Despite the lack of a reduction in CFUs in the group that received the combined treatment, there was a significant reduction in the size of the lesions in the lungs after 70 days of infection. At the same time, the IL-10 levels were higher in the treated mice than in the infected-only mice. Moreover, significant levels of rPb40-specific IgG antibodies were detected in the sera of immunized mice. Thus, the treatment protocol consisting of rPb40 immunization in addition to fluconazole chemotherapy showed an additive protective effect after intratracheal challenge, preventing fungal dissemination to other sites of infection and preventing relapses. These results provide new prospects for PCM immunotherapy.


Assuntos
Adjuvantes Imunológicos/administração & dosagem , Antifúngicos/administração & dosagem , Antígenos de Fungos/administração & dosagem , Paracoccidioidomicose/tratamento farmacológico , Adjuvantes Imunológicos/genética , Adjuvantes Imunológicos/isolamento & purificação , Animais , Anticorpos Antifúngicos/sangue , Antígenos de Fungos/genética , Antígenos de Fungos/isolamento & purificação , Tratamento Farmacológico/métodos , Fluconazol/administração & dosagem , Imunoglobulina G/sangue , Imunoterapia/métodos , Interleucina-10/sangue , Fígado/microbiologia , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Baço/microbiologia , Resultado do Tratamento
16.
Acta Physiol (Oxf) ; 205(2): 292-301, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22168399

RESUMO

AIM: To examine whether physical activity increases osteogenic differentiation of bone marrow mesenchymal stem cells (BMMSCs) from adult rats compared with young rats. METHODS: Eighteen female Wistar rats were divided into three groups and the following cells isolated: (1) differentiated BMMSCs from young donors, (2) differentiated BMMSCs from sedentary adult donors and (3) differentiated BMMSCs from active adult donors. We analysed MTT conversion, percentage of cells per field, mineralized nodule number and gene expression for telomerase reverse transcriptase (TERT), alkaline phosphatase, caspase 3, osteocalcin, bone sialoprotein and collagen I. RESULTS: Telomerase reverse transcriptase expression and the percentage of cells per field in BMMSCs cultures from adult rats were smaller than those observed in young donors. However, levels of caspase 3 expression were higher in BMMSCs from adult donors (P < 0.05). Despite the fact that physical activity was associated with an increase in expression of caspase 3 (P < 0.05), there was no difference in the percentage of cells per field between groups of adult BMMSCs (active or sedentary). However, physical activity increased the number of mineralized nodules and osteocalcin expression after 21 days, and alkaline phosphatase expression at 7, 14 and 21 days in the BMMSCs of adult donors (P < 0.05). However, those values were smaller when compared with young donors BMMSCs (P < 0.05). Only the expression levels of alkaline phosphatase were similar to young donors BMMSCs (P ≥ 0.05). CONCLUSION: Physical activity increases osteogenic differentiation of BMMSCs from adult donors but does not increase the differentiation to the levels observed in BMMSCs from young donor rats.


Assuntos
Envelhecimento/fisiologia , Células da Medula Óssea/fisiologia , Células-Tronco Mesenquimais/fisiologia , Osteogênese/fisiologia , Condicionamento Físico Animal/fisiologia , Animais , Células da Medula Óssea/citologia , Células Cultivadas , Feminino , Células-Tronco Mesenquimais/citologia , Ratos , Ratos Wistar
17.
J Immunol Methods ; 367(1-2): 78-84, 2011 Mar 31.
Artigo em Inglês | MEDLINE | ID: mdl-21354162

RESUMO

Paracoccidioidomycosis (PCM) is one of the most important endemic mycoses in Latin America; it's usually diagnosed by observation and/or isolation of the etiologic agent, Paracoccidioides brasiliensis, as well as by a variety of immunological methods, such as complement fixation and immunodiffusion. Although these approaches are useful, historically their sensitivity and specificity have often been compromised by the use of complex mixtures of undefined antigens. The use of combinations of purified, well-characterized antigens appears preferable and may yield good results. In the present study combinations of the previously described 27-kDa recombinant antigen (rPb27) and a recombinant 40-kDa-molecular-mass antigen (rPb40) from this fungus, that was identified by Goes et al. (2005) through the AST strategy as a homolog of Neurospora crassa calcineurin B, were used in an indirect enzyme linked immunosorbent assay (ELISA) for diagnosis and follow-up of patients with PCM. The complete coding cDNA of rPb40 and rPb27 were cloned into a pET-21a and a pET-DEST 42 plasmid, respectively, expressed in E. coli with a his-tag and purified by affinity chromatography. Among 109 PCM serum samples analyzed, a homogeneous IgG response to these proteins was observed. 62 serum samples from patients with other diseases, 18 from patients with other mycosis and 23 from healthy individuals were also studied. Detection of anti-rPb27 and anti-rPb40 antibodies in sera of patients with PCM by ELISA using a combination of the two purified proteins showed a sensitivity of 96% with a specificity of 100% in relation to control normal human sera and to sera from patients with other systemic mycosis and 93.5% to sera from patients with diverse infections. The use of this two proteins combination provided an excellent immunodiagnosis assay with great values of sensitivity and specificity, even in relation to sera from patients with other mycosis, making possible the standardization of a new methodology to diagnose this important mycosis, with a good confiability and reprodutibility.


Assuntos
Anticorpos Antifúngicos/sangue , Antígenos de Fungos/imunologia , Ensaio de Imunoadsorção Enzimática/métodos , Paracoccidioides/imunologia , Paracoccidioidomicose/diagnóstico , Adolescente , Adulto , Idoso , Humanos , Imunoglobulina G/sangue , Pessoa de Meia-Idade , Paracoccidioidomicose/imunologia , Proteínas Recombinantes/imunologia , Sensibilidade e Especificidade
18.
Oncol Rep ; 24(3): 677-85, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20664973

RESUMO

Breast cancer is a major health burden, responsible for >10% of all cases of cancer worldwide. Advances in breast cancer diagnosis and treatment have contributed to an improved rate of survival, although mortality rates remains significantly high. The establishment of breast cancer cell lines is an important model for understanding biological processes involved in this disease and for identifying potential therapeutic targets. The novel human breast cancer cell lines, MACL-1 and MGSO-3, were used in this study to identify possible surface antigens by antibodies directed against two commercial breast cancer cell lines MCF-7 and MDA-MB-231. We purified a 37 kDa antigen by affinity chromatography from MDA-MB-231, and its N-terminal amino acid sequence was homologous to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Therefore, immunohistochemical experiments, using specific monoclonal antibodies, evidenced a co-localization of GAPDH and Na+/K+-ATPase on the surface of commercially available and recently established breast cancer cell lines. It is of note that Na+/K+-ATPase was used as a plasma membrane marker. This finding opens new perspectives for breast cancer diagnosis and treatment since GAPDH could be used as a biomarker or as a potential therapeutic target in breast cancer.


Assuntos
Antígenos de Neoplasias/isolamento & purificação , Biomarcadores Tumorais/isolamento & purificação , Neoplasias da Mama/enzimologia , Gliceraldeído-3-Fosfato Desidrogenases/isolamento & purificação , Proteínas de Membrana/isolamento & purificação , Neoplasias da Mama/imunologia , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Proliferação de Células , Forma Celular , Sobrevivência Celular , Cromatografia de Afinidade , Feminino , Humanos , Imuno-Histoquímica , Análise de Sequência de Proteína , ATPase Trocadora de Sódio-Potássio/isolamento & purificação
19.
Cytotherapy ; 12(4): 478-90, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20230220

RESUMO

BACKGROUND AIMS: Stem cells derived from human adipose tissue (ASC) have the capacity for renewal, are easily obtained and have plasticity properties that allow them to differentiate into several cell types, including osteoblast cells. With the aim of understanding the issue of the osteogenic process and finding reliable biomarkers in cells undergoing the osteogeneic differentiation process, this work took advantage of a proteomic approach to identify proteins involved in osteogenesis. METHODS: For this purpose, ASC were analyzed under three conditions: S0, in the absence of stimulation; S1, with 2 weeks of osteogenic medium stimulation; and S2, with 4 weeks of osteogenic medium stimulation. The identification of ASC was carried out by flow cytometry using antibodies specific to known undifferentiated stem cell-surface markers. Cell viability, enzymatic activity, mineral deposition, collagen structure and production and gene analyzes were evaluated for each condition. RESULTS: Phenotypic modifications were observed during the in vitro osteogenic differentiation process by two-dimensional (2-D) differential image gel electrophoresis (DIGE). The proteins were identified by mass espectrometry in tandem (MS/MS) analyzes using Matrix-assisted laser desorption/ionization with TOF/TOF is a tandem mass spectrometry method where two time-of-flight mass spectrometers are used consecutively (MALDI-TOF/TOF). A total of 51 differentially expressed proteins was identified when comparing the three observed conditions. Sixteen different spots were identified in the S0 stage compared with S2, while 28 different spots were found in S2 compared with S0. S1 expressed seven different spots compared with S0 and S2. CONCLUSIONS: These findings suggest the involvement of several proteins directly related to the osteogenic pathway, which can be used to improve understanding of the osteogenic process.


Assuntos
Tecido Adiposo/citologia , Biomarcadores , Células-Tronco Mesenquimais/metabolismo , Osteoblastos/metabolismo , Proteômica , Células Estromais/metabolismo , Tecido Adiposo/cirurgia , Adulto , Biomarcadores/metabolismo , Diferenciação Celular , Separação Celular , Células Cultivadas , Eletroforese em Gel Bidimensional , Feminino , Citometria de Fluxo , Perfilação da Expressão Gênica , Humanos , Células-Tronco Mesenquimais/citologia , Pessoa de Meia-Idade , Osteoblastos/citologia , Proteômica/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Células Estromais/citologia
20.
Clin Exp Immunol ; 160(2): 266-74, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20132231

RESUMO

Schistosoma mansoni infection has been associated with protection against allergies. The mechanisms underlying this association may involve regulatory cells and cytokines. We evaluated the immune response induced by the S. mansoni antigens Sm22.6, PIII and Sm29 in a murine model of ovalbumin (OVA)-induced airway inflammation. BALB/c mice were sensitized with subcutaneously injected OVA-alum and challenged with aerolized OVA. Mice were given three doses of the different S. mansoni antigens. Lung histopathology, cellularity of bronchoalveolar lavage (BAL) and eosinophil peroxidase activity in lung were evaluated. Immunoglobulin (Ig)E levels in serum and cytokines in BAL were also measured. Additionally, we evaluated the frequency of CD4+forkhead box P3 (FoxP3)+ T cells in cultures stimulated with OVA and the expression of interleukin (IL)-10 by these cells. The number of total cells and eosinophils in BAL and the levels of OVA-specific IgE were reduced in the immunized mice. Also, the levels of IL-4 and IL-5 in the BAL of mice immunized with PIII and Sm22.6 were decreased, while the levels of IL-10 were higher in mice immunized with Sm22.6 compared to the non-immunized mice. The frequency of CD4+FoxP3+ T cells was higher in the groups of mice who received Sm22.6, Sm29 and PIII, being the expression of IL-10 by these cells only higher in mice immunized with Sm22.6. We concluded that the S. mansoni antigens used in this study are able to down-modulate allergic inflammatory mediators in a murine model of airway inflammation and that the CD4+FoxP3+ T cells, even in the absence of IL-10 expression, might play an important role in this process.


Assuntos
Alveolite Alérgica Extrínseca/imunologia , Antígenos de Helmintos/imunologia , Schistosoma mansoni/imunologia , Alveolite Alérgica Extrínseca/induzido quimicamente , Alveolite Alérgica Extrínseca/prevenção & controle , Animais , Asma , Líquido da Lavagem Broncoalveolar/química , Linfócitos T CD4-Positivos/química , Linfócitos T CD4-Positivos/imunologia , Citocinas/análise , Modelos Animais de Doenças , Feminino , Fatores de Transcrição Forkhead/análise , Imunização , Interleucinas/análise , Pulmão/imunologia , Pulmão/patologia , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/administração & dosagem , Ovalbumina/toxicidade , Eosinofilia Pulmonar/induzido quimicamente , Eosinofilia Pulmonar/imunologia , Eosinofilia Pulmonar/prevenção & controle , Subpopulações de Linfócitos T/química , Subpopulações de Linfócitos T/imunologia
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