RESUMO
Solanum glaucophyllum Desf. is a calcinogenic plant responsible for enzootic calcinosis that affects ruminants and causes alterations in bone and cartilaginous tissues, among others. It is believed that changes in cartilage tissue, with reduced bone growth, are due to hypercalcitoninism, caused by excess vitamin D. However, we hypothesized that S. glaucophyllum Desf. can act directly on chondrocytes and therefore, chondrocyte cultures from the epiphysis of the long bones of newborn rats were used as a model to elucidate the direct effects of S. glaucophyllum Desf. on bone growth. Plant samples were collected from Cañuelas, Argentina. An aliquot of the plant extract was used to quantify vitamin D (1,25(OH)2D3). The effects of the three concentrations of the plant extract were tested in cultures of chondrocytes extracted from the epiphyses of the long bones of 32 three-day-old Wistar rats. A control group (without extract), and three groups treated with different concentrations of plant extract were formed: group 1 (100 µL/L); group 2 (1 mL/L), and group 3 (5 mL/L), containing respectively 1 × 10-9 M, 1 × 10-8 M, and 5 × 10-8 M of 1,25(OH)2D3. After 7, 14, and 21 days of culture, MTT assay for cell viability, alkaline phosphatase activity, and quantification of the percentage of areas with glycosaminoglycans (GAG) stained with periodic acid-Schiff (PAS) were performed. On day 7, all chondrocytes in group 3, that is, those with the highest concentration of plant extract, died. On days 14 and 21, groups 1 and 2 showed a significant reduction in chondrocyte viability compared to the control. At 7, 14, and 21 days, groups 1 and 2 showed significantly lower alkaline phosphatase activity than the control. On day 21, group 2 showed a significant reduction in areas with PAS + GAGs. There were no significant differences between the groups in the expression of gene transcripts for Sox9, Col2, ColX, and aggrecan. The S. glaucophyllum Desf. extract directly affected growing rat chondrocytes by reducing viability, alkaline phosphatase activity, and GAG synthesis without altering the expression of gene transcripts for Sox9, Col2, ColX, and aggrecan, which may be one of the mechanisms by which there is a reduction in bone growth in animals intoxicated by the plant.
Assuntos
Condrócitos , Solanum glaucophyllum , Ratos , Animais , Condrócitos/metabolismo , Animais Recém-Nascidos , Calcitriol/metabolismo , Ratos Wistar , Agrecanas/metabolismo , Fosfatase Alcalina , Cartilagem , Plantas , Vitamina D/metabolismo , Extratos Vegetais , Células CultivadasRESUMO
Decidual immunological mediators modulate placental formation, decidualization and fetal development. However, the effect of maternal hyperthyroidism on decidual immunology needs further research. The aim of this study was to evaluate the population of uterine natural killer cells (uNKs) and the expression of immunological mediators in the decidua of female rats throughout pregnancy. Wistar rats were used and hyperthyroidism was induced by daily administration of L-thyroxine (T4) throughout pregnancy. The population of uNK cells in decidua was evaluated by immunostaining Lectin DBA, as well as the expression of interferon γ (INFγ), macrophage migration inhibitory factor (MIF), interleukin 15 (IL-15) and inducible nitric oxide synthase (iNOS) at 7, 10, 12, 14 and 19 days of gestation (DG). Maternal hyperthyroidism reduced the DBA+ uNK cell population in the decidua at 7 (P < 0.05) and 10 (P < 0.01) DGs compared to that in the control group, while it increased in the basal decidua (P < 0.05) and metrial gland (P < 0.0001) at the 12th DG. Hyperthyroidism also increased immunostaining of IL-15 (P < 0.0001), INFγ (P < 0.05), and MIF (P < 0.05) in the 7th DG, and increased immunostaining of IL-15 (P < 0.0001) and MIF (P < 0.01) in the 10th DG. However, excess thyroxine reduced IL-15 expression in the metrial gland and/or basal decidua in the 12th (P < 0.05), 14th (P < 0.01), and 19th (P < 0.001) DGs, as was also observed for INFγ in the basal decidua (P<0.001) and metrial gland (P < 0.0001) in the 12th DG. Regarding iNOS, an antiinflammatory cytokine, lower expression was observed in the basal decidua of hyperthyroid animals at 7 and 12 DGs (P < 0.05), whereas an increase occurred in the 10th DG (P < 0.05). These data demonstrate that maternal hyperthyroidism in female rats, particularly between 7 and 10 DGs, reduces the population of DBA+ uNKs in the decidua and increases the expression of inflammatory cytokines, suggesting a more proinflammatory environment in early pregnancy caused by this gestational disease.
Assuntos
Hipertireoidismo , Placenta , Ratos , Gravidez , Feminino , Animais , Placenta/metabolismo , Decídua/metabolismo , Interleucina-15/metabolismo , Interleucina-15/farmacologia , Ratos Wistar , Células Matadoras Naturais/metabolismo , Hipertireoidismo/metabolismoRESUMO
OBJECTIVE: We sought to evaluate the effect of different concentrations of ethanol on phenotype and activity of articular chondrocyte synthesis of neonatal rats in 2-dimensional (2D) and 3-dimensional (3D) culture. METHODS: Chondrocytes were cultured in chondrogenic medium with different concentrations of ethanol: 0.0% v/v (control); 0.05% v/v (8.6 mM); 0.25% v/v (42.9 mM), and 0.5% v/v (85.7 mM). Chondrocytes under 2D culture were subjected to MTT assay, while chondrocytes under 3D culture were processed for paraffin inclusion and stained by periodic acid Schiff (PAS) to evaluate mean chondrocyte diameter and percentages of cells, nucleus, cytoplasm, well-differentiated matrix, and PAS+ areas. The expression of gene transcripts for aggrecan, Sox9, and type II collagen was evaluated by real-time quantitative polymerase chain reaction. RESULTS: There was no difference between groups by the MTT assay. PAS staining revealed that chondrocytes treated with 0.5% v/v ethanol had higher percentages of cytoplasm and nuclear areas, but with a reduction in PAS+ matrix area. The mean diameter of chondrocytes was similar between groups. The expression of aggrecan in the group treated with 0.5% v/v ethanol was lower in comparison to that in the control. In the groups treated with 0.25% v/v and 0.5% v/v ethanol, the percentage of differentiated cartilage was lower in comparison with that in the control. The group treated with 0.05% v/v ethanol was similar to the control in all parameters. CONCLUSIONS: Ethanol acted directly on in vitro cultured articular chondrocytes of newborn rats, altering the chondrocyte phenotype and its synthesis activity, and these effects were dose dependent.
Assuntos
Cartilagem Articular , Condrócitos , Animais , Cartilagem Articular/metabolismo , Condrócitos/metabolismo , Colágeno Tipo II/metabolismo , Etanol/metabolismo , Etanol/farmacologia , Fenótipo , RatosRESUMO
The Solanum glaucophyllum Desf. has been used to treat and prevent diseases in human and veterinary medicine. On the other hand, plant poisoning causes several bone diseases, among them osteoporosis, which is characterized by osteoblastic hypoplasia. Because the osteoblast is a cell derived from the differentiation of mesenchymal stem cells (MSCs) from bone marrow, the hypothesis is that the plant reduces the osteogenic differentiation of MSCs. The objective of this study was to evaluate the effects of S. glaucophyllum Desf. extract on MSCs cultured in osteogenic differentiation medium. We determined by liquid chromatography that 1 ml of plant extract contained 3.8 µl of 1,25(OH)2 D3 (calcitriol). Four groups of MSCs cultivated in osteogenic medium were evaluated as follows: (a) treated with 100 µl of extract/L containing 0.4 µg/L of calcitriol; (b) treated with 1 ml of extract/L containing 4 µg/L of calcitriol; (c) treated with 5 ml of extract/L containing 20 µg/L of calcitriol; and (d) a control group without extract. We performed alkaline phosphatase activity assay, analysis of MTT conversion to formazan, and evaluated the percentage of cells, and number and diameter of mineralization nodules. The expression of gene transcripts for osteopontin, bone sialoprotein and BMP-2 was analysed by RT-qPCR. After 21 days, there was a significant reduction in MTT conversion to formazan in treated groups, of the cellularity in the group with 5 ml of extract/L, and in the number and size of mineralization nodules in the groups treated with 1 and 5 ml of extract/L. The 5 ml extract/L concentration also reduced transcript expression of osteopontin. It is concluded that S. glaucophyllum Desf. at concentrations of 1 and 5 ml extract/L reduced mineralized matrix synthesis in MSCs cultivated in osteogenic differentiation medium, which suggests that this is one of the mechanisms by which osteoporosis occurs in intoxicated animals.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Extratos Vegetais/farmacologia , Solanum glaucophyllum/química , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Animais , Proteína Morfogenética Óssea 2/genética , Proteína Morfogenética Óssea 2/metabolismo , Diferenciação Celular/fisiologia , Relação Dose-Resposta a Droga , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Sialoproteína de Ligação à Integrina/genética , Sialoproteína de Ligação à Integrina/metabolismo , Células-Tronco Mesenquimais/fisiologia , Osteopontina/genética , Osteopontina/metabolismo , Extratos Vegetais/administração & dosagem , Extratos Vegetais/química , RatosRESUMO
The hypothesis of this experiment is that mesenchymal stem cells (MSCs) are involved in the genesis of the bone metaplasia caused by Solanum glaucophyllum intoxication. We determined using liquid chromatography that 1â¯mL of plant extract contained 3.8⯵l of 1,25(OH)2D3. The ability of 100⯵L, 1â¯mL and 5â¯mL of extract/L, containing 1â¯nM (0.4⯵g/L), 10â¯nM (4⯵g/L) and 50â¯nM (20⯵g/L) of 1,25(OH)2D3, respectively, in inducing the osteogenic differentiation in bone marrow MSCs from rats was tested. At the concentrations of 1 and 5â¯mL of extract/L of culture medium without osteogenesis-inducing factors, the plant extract induced the osteogenic differentiation of the MSCs, as was evidenced by the greater synthesis of mineralized matrix. At the higher concentration (5â¯mL of extract/L), an increase in the relative expression of BMP-2 gene was observed. It was concluded that rat bone marrow MSC culture is a good model for studying the effects of the S. glaucophyllum extract on the osteogenic differentiation of undifferentiated cells. Also, S. glaucophyllum extracts containing 10â¯nM (4⯵g/L) and 50â¯nM (20⯵g/L) of 1,25(OH)2D3 induce the osteogenic differentiation of MSCs, suggesting that this is one of the mechanisms by which S. glaucophyllum causes bone metaplasia.
Assuntos
Células-Tronco Mesenquimais/efeitos dos fármacos , Metaplasia/induzido quimicamente , Extratos Vegetais/toxicidade , Solanum glaucophyllum/química , Animais , Proteína Morfogenética Óssea 2/metabolismo , Osso e Ossos/patologia , Cromatografia Líquida , Sialoproteína de Ligação à Integrina/metabolismo , Células-Tronco Mesenquimais/patologia , Osteopontina/metabolismo , Ratos , Testes de ToxicidadeRESUMO
Objetivou-se estabelecer um protocolo para extração, cultivo e expansão de células tronco mesenquimais (CTM), utilizando-se 3,0 mL da medula óssea e 3,0 cm3 de tecido adiposo do subcutâneo de três cães machos com seis meses de idade. As amostras foram processadas e as células extraídas e cultivadas em DMEM. Para comprovação do isolamento de CTM, procedeu-se a caracterização fenotípica e a diferenciação osteogênica, adipogênica e condrogênica. As células isoladas apresentaram morfologia alongada e fusiforme e capacidade de se diferenciar em osteoblastos, adipócitos e condrócitos. A caracterização fenotípica revelou alta expressão de marcadores de CTM CD90 (80,04%) e CD29 (96%) nas células de origem medular e CD90 (60,94%) e CD29 (77,08%) nas de origem adiposa. A expressão de marcadores hematopoiéticos foi baixa tanto nas células de origem medular CD45 (1,45%) e CD34 (1,53%), quanto nas de origem adiposa CD45 (1,45%) e CD34 (1,53%). As modificações e adaptações realizadas nos protocolos clássicos simplificaram o processo e foram eficientes, permitindo o isolamento e cultivo de CTM da medula óssea e do tecido adiposo de cães. (AU)
The objective of this study was to establish a protocol for the isolation and culture of mesenchymal stem cells (MSC) from bone marrow and adipose tissue of dogs. Three 6-month-old male dogs were used. Approximately 3.0 cm3 of adipose tissue and 3.0 mL of bone marrow were collected. The samples were processed and the isolated cells were cultured in DMEM. The cells were subjected to phenotypic characterization and to osteogenic, adipogenic, and condrogenic differentiation to confirm the isolation of the MSC. The cells showed elongated and fusiform morphology and they were able to differentiate into osteoblasts, adipocytes, and chondrocytes. Phenotypic characterization revealed high expression of the MSC markers CD90(80.04%) and CD29(96%) in the cells from bone marrow and high expression of CD90(60.94%) and CD29(77.08%) in the cells from adipose tissue. In addition, phenotypic characterization revealed low expression of hematopoietic markers CD45(1.45%) and CD34(1.53%) in the cells from bone marrow and low expression of CD45(1.45%) and CD34(1.53%) in the cells from adipose tissue. Based on these results, the modifications applied to classical protocols simplified the process and proved to be efficient in the isolation, culture, and expansion of MSC isolated from the bone marrow and adipose tissue of dogs.(AU)
Assuntos
Animais , Cães , Células-Tronco , Gordura Subcutânea , Medula Óssea , Técnicas de Cultura de Células/veterinária , Diferenciação CelularRESUMO
Objetivou-se estabelecer um protocolo para extração, cultivo e expansão de células tronco mesenquimais (CTM), utilizando-se 3,0 mL da medula óssea e 3,0 cm3 de tecido adiposo do subcutâneo de três cães machos com seis meses de idade. As amostras foram processadas e as células extraídas e cultivadas em DMEM. Para comprovação do isolamento de CTM, procedeu-se a caracterização fenotípica e a diferenciação osteogênica, adipogênica e condrogênica. As células isoladas apresentaram morfologia alongada e fusiforme e capacidade de se diferenciar em osteoblastos, adipócitos e condrócitos. A caracterização fenotípica revelou alta expressão de marcadores de CTM CD90 (80,04%) e CD29 (96%) nas células de origem medular e CD90 (60,94%) e CD29 (77,08%) nas de origem adiposa. A expressão de marcadores hematopoiéticos foi baixa tanto nas células de origem medular CD45 (1,45%) e CD34 (1,53%), quanto nas de origem adiposa CD45 (1,45%) e CD34 (1,53%). As modificações e adaptações realizadas nos protocolos clássicos simplificaram o processo e foram eficientes, permitindo o isolamento e cultivo de CTM da medula óssea e do tecido adiposo de cães.
The objective of this study was to establish a protocol for the isolation and culture of mesenchymal stem cells (MSC) from bone marrow and adipose tissue of dogs. Three 6-month-old male dogs were used. Approximately 3.0 cm3 of adipose tissue and 3.0 mL of bone marrow were collected. The samples were processed and the isolated cells were cultured in DMEM. The cells were subjected to phenotypic characterization and to osteogenic, adipogenic, and condrogenic differentiation to confirm the isolation of the MSC. The cells showed elongated and fusiform morphology and they were able to differentiate into osteoblasts, adipocytes, and chondrocytes. Phenotypic characterization revealed high expression of the MSC markers CD90(80.04%) and CD29(96%) in the cells from bone marrow and high expression of CD90(60.94%) and CD29(77.08%) in the cells from adipose tissue. In addition, phenotypic characterization revealed low expression of hematopoietic markers CD45(1.45%) and CD34(1.53%) in the cells from bone marrow and low expression of CD45(1.45%) and CD34(1.53%) in the cells from adipose tissue. Based on these results, the modifications applied to classical protocols simplified the process and proved to be efficient in the isolation, culture, and expansion of MSC isolated from the bone marrow and adipose tissue of dogs.
Assuntos
Animais , Cães , Células-Tronco , Gordura Subcutânea , Medula Óssea , Diferenciação Celular , Técnicas de Cultura de Células/veterináriaRESUMO
Abstract The objective of this study was to establish a protocol for the isolation and culture of mesenchymal stem cells (MSC) from bone marrow and adipose tissue of dogs. Three 6-month-old male dogs were used. Approximately 3.0 cm3 of adipose tissue and 3.0 mL of bone marrow were collected. The samples were processed and the isolated cells were cultured in DMEM. The cells were subjected to phenotypic characterization and to osteogenic, adipogenic, and condrogenic differentiation to confirm the isolation of the MSC. The cells showed elongated and fusiform morphology and they were able to differentiate into osteoblasts, adipocytes, and chondrocytes. Phenotypic characterization revealed high expression of the MSC markers CD90(80.04%) and CD29(96%) in the cells from bone marrow and high expression of CD90(60.94%) and CD29(77.08%) in the cells from adipose tissue. In addition, phenotypic characterization revealed low expression of hematopoietic markers CD45(1.45%) and CD34(1.53%) in the cells from bone marrow and low expression of CD45(1.45%) and CD34(1.53%) in the cells from adipose tissue. Based on these results, the modifications applied to classical protocols simplified the process and proved to be efficient in the isolation, culture, and expansion of MSC isolated from the bone marrow and adipose tissue of dogs.
Resumo Objetivou-se estabelecer um protocolo para extração, cultivo e expansão de células tronco mesenquimais (CTM), utilizando-se 3,0 mL da medula óssea e 3,0 cm3 de tecido adiposo do subcutâneo de três cães machos com seis meses de idade. As amostras foram processadas e as células extraídas e cultivadas em DMEM. Para comprovação do isolamento de CTM, procedeu-se a caracterização fenotípica e a diferenciação osteogênica, adipogênica e condrogênica. As células isoladas apresentaram morfologia alongada e fusiforme e capacidade de se diferenciar em osteoblastos, adipócitos e condrócitos. A caracterização fenotípica revelou alta expressão de marcadores de CTM CD90 (80,04%) e CD29 (96%) nas células de origem medular e CD90 (60,94%) e CD29 (77,08%) nas de origem adiposa. A expressão de marcadores hematopoiéticos foi baixa tanto nas células de origem medular CD45 (1,45%) e CD34 (1,53%), quanto nas de origem adiposa CD45 (1,45%) e CD34 (1,53%). As modificações e adaptações realizadas nos protocolos clássicos simplificaram o processo e foram eficientes, permitindo o isolamento e cultivo de CTM da medula óssea e do tecido adiposo de cães.
RESUMO
The objective of the present study was to evaluate the effect of the thyroid hormones in the gene transcription and immunohistochemical expression of hormonal and angiogenic factors in the placenta of rats. Seventy-two adult female rats were divided equally into propylthiouracil (PTU)-treated, thyroxine (T4)-treated, and control groups. The animals were sacrificed at 10, 14, and 19 days of gestation. We evaluated the immunohistochemical expression of VEGF and its receptor Flk-1. The gene transcription of VEGF, Flk-1, PGF, sFlt1, PL-1, and rPlf was evaluated in placental discs by real-time RT-PCR. The data were analyzed using a Student-Newman-Keuls (SNK) test. At day 10, T4-treated rats presented increased VEGF and PGF gene expression, while PTU-treated rats showed increased rPlf gene expression. Both groups showed reduced Flk-1 and PL-1 gene expression at day 10. At day 14, PTU-treated rats showed reduced VEGF, PGF, and rPlf gene expression. PTU-treated group showed reduced VEGF immunostaining in the placental labyrinth at 14 and 19 days of gestation but it showed increased VEGF immunostaining in the spongiotrophoblast layer at day 14. PTU-treated rats showed increased Flk-1 expression at 14 days of gestation. At days 14 and 19, T4-treated group showed increased PL-1 gene expression and reduced VEGF immunostaining. T4-treated rats also showed reduced Flk-1 and sFlt-1 expression at day 19. Both groups showed increased rPlf gene expression at day 19. In conclusion, rats treated with PTU and T4 have differential effects on the expression of factors involved in placental angiogenic and hormonal activity, and these effects are dependent on the gestational period.
Assuntos
Antitireóideos/farmacologia , Placenta/efeitos dos fármacos , Propiltiouracila/farmacologia , Tiroxina/farmacologia , Fator A de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Animais , Feminino , Placenta/metabolismo , Fator de Crescimento Placentário , Lactogênio Placentário/genética , Lactogênio Placentário/metabolismo , Gravidez , Proteínas da Gravidez/genética , Proteínas da Gravidez/metabolismo , Ratos , Fator A de Crescimento do Endotélio Vascular/genética , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/genética , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/genéticaRESUMO
The objective of the present study was to evaluate the gene and immunohistochemical expression of inflammatory mediators involved in the immune activity and the intrauterine trophoblast migration of the placentas in hypothyroid and L-thyroxine (L-T4)-treated rats. A total of 144 adult female rats were divided equally into hypothyroid, l-T4-treated, and euthyroid (control) groups. Hypothyroidism was induced by daily administration of propylthiouracil. Rats were killed at 0, 10, 14, 15, 16, 17, 18, and 19 days of gestation. We evaluated the depth of interstitial and endovascular intrauterine trophoblast invasion and the immunohistochemical expression of interferon γ (INFy), migration inhibitory factor (MIF), and inducible nitric oxide synthase (NOS2 (iNOS)). The gene expression of Toll-like receptor 2 (Tlr2) and Tlr4, Infy, Mif, tumor necrosis factor (Tnf (Tnfα)), Il10, Nos2, matrix metalloproteinase 2 (Mmp2) and Mmp9, and placental leptin was also measured in placental disks by real-time RT-PCR. The data were analyzed using an Student-Newman-Keuls (SNK) test. Hypothyroidism reduced the endovascular and interstitial trophoblast migration, and the expression of TLR4, INFy, MIF, interleukin 10 (IL10), NOS2, MMP2 and MMP9, and placental leptin, while increased the expression of TLR2 (P<0.05). T4-treated rats not only increased the expression of IL10 and NOS2 but also reduced the expression of TNF and MIF at 10 days of gestation (P<0.05). However, at 19 days of gestation, expression of INFy and MIF was increased in T4-treated group (P<0.05). Excess of T4 also increased the gene expression of Mmp2 at 10 days of gestation (P<0.05), but reduced the endovascular trophoblast migration at 18 days of gestation (P<0.05). Hypothyroidism and excess of T4 differentially affect the immune profile and the intrauterine trophoblast migration of the placenta, and these effects are dependent on the gestational period.
Assuntos
Movimento Celular , Hipertireoidismo/metabolismo , Hipotireoidismo/metabolismo , Mediadores da Inflamação/metabolismo , Placenta/metabolismo , Glândula Tireoide/metabolismo , Trofoblastos/metabolismo , Útero/metabolismo , Animais , Modelos Animais de Doenças , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Idade Gestacional , Hipertireoidismo/genética , Hipertireoidismo/imunologia , Hipertireoidismo/fisiopatologia , Hipotireoidismo/genética , Hipotireoidismo/imunologia , Hipotireoidismo/fisiopatologia , Cinética , Placenta/fisiopatologia , Gravidez , RNA Mensageiro/metabolismo , Ratos Wistar , Glândula Tireoide/fisiopatologia , Útero/fisiopatologiaRESUMO
The aim of this work was to evaluate the mesenchymal stem cells treatment of rats with myonecrosis caused by Rhinocerophis alternatus venom through acute phase proteins (APP) profile. The animals were distributed into three experimental groups (G1, G2 and G3). G1 and G2 were inoculated with 120 μg of R. alternatus venom diluted in 200 µL of ultra-pure water in gastrocnemic muscle, while G3 received 200 µL of ultra-pure water. Three days after, G1 was treated with 5 X 10(6) MSC diluted in PBS and G2 and G3 only with PBS. Each three days after the treatments (3rd, 6th, 9th, 12th 15th days), blood of five animals in each group was collected in order to evaluate the APP. A decrease (P<0.05) in α2-globulin fraction was observed in G1 on the 6th day. In G1 and G2, a raise (P<0.05) was observed in β globulin, a common occurrence in the late phases of inflammatory process, although no significant difference was observed between them. Concerning gamma globulins levels, on the 6th day after the treatments, in G1 and G2 groups, increase in the levels was observed. These data showed that the MSC treatment after bothropic envenomation in the rats caused alteration in APP.
RESUMO
OBJECTIVE: To examine if triiodothyronine (T3) increases osteogenic differentiation adipose tissue derived stem cells (ASCs) from ovariectomized adult rats with osteoporosis compared with young rats and adult rats without osteoporosis. MATERIALS AND METHODS: The ASCs were cultured in osteogenic medium and distributed into seven groups: 1) ASCs of young rats without osteoporosis; 2) ASCs of adult rats without osteoporosis; 3) ASCs of adult rats with osteoporosis and 4, 5, 6 and 7) ASCs of adult rats with osteoporosis treated with T3 (0.01 nM, 1 nM, 100 nM and 1,000 nM). We analyzed alkaline phosphatase activity, dimethylthiazol (MTT) conversion, percentage of mineralized nodules, cellularity and quantification of gene transcripts for collagen I, osteocalcin, osteopontin and Bmp-2. RESULTS: Regardless of the dose, T3 reduced the MTT conversion, alkaline phosphatase activity, percentage of cells and the expression of collagen I in at least one of the doses and periods studied (p < 0.05). But, the treatment with T3 does not modify the number of mineralized nodules and the expression of osteopontin and Bmp-2 in culture of ASCs from adult rats with osteoporosis (p > 0.05). CONCLUSION: T3 has a negative effect on some factors involved in osteogenic differentiation of ASCs from adult rats with osteoporosis, without; however, reduce the formation of mineralized nodules and the expression of bone proteins.
Assuntos
Tecido Adiposo/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Osteoporose , Tri-Iodotironina/farmacologia , Fatores Etários , Fosfatase Alcalina/metabolismo , Animais , Diferenciação Celular , Feminino , Células-Tronco Mesenquimais/enzimologia , Células-Tronco Mesenquimais/fisiologia , Osteogênese/fisiologia , Osteoporose/patologia , Ovariectomia , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo RealRESUMO
OBJETIVO: Avaliar se a triiodotironina (T3) aumenta a diferenciação osteogênica das células-tronco mesenquimais do tecido adiposo (CTM-TA) de ratas adultas ovariectomizadas e com osteoporose e compará-lo ao de ratas adultas e jovens sem osteoporose. MATERIAIS E MÉTODOS: CTM-TA foram cultivadas em meio osteogênico e distribuídas em sete grupos: 1) CTM-TA de ratas jovens sem osteoporose; 2) CTM-TA de ratas adultas sem osteoporose; 3) CTM-TA de ratas adultas com osteoporose e 4, 5, 6 e 7) CTM-TA de ratas adultas com osteoporose tratadas com T3 (0,01 nM, 1 nM, 100 nM e 1.000 nM). AVALIARAM-SE: atividade da fosfatase alcalina, conversão do dimetiltiazol (MTT), porcentagem de nódulos de mineralização, celularidade e quantificação de transcriptos gênicos para colágeno I, osteocalcina, osteopontina e Bmp-2. RESULTADOS: Independente da dose, T3 reduziu a conversão do MTT, a atividade da fosfatase, a porcentagem de células e a expressão de colágeno I em pelo menos uma das doses e dos períodos estudados (p < 0,05). Mas o tratamento com T3 não alterou o número de nódulos de mineralização e a expressão de osteopontina e Bmp-2 em culturas de CTM-TA de ratas adultas com osteoporose (p > 0,05). CONCLUSÃO: T3 apresenta efeitos negativos sobre alguns fatores envolvidos na diferenciação osteogênica de CTM-TA, sem, no entanto, reduzir a formação de nódulos de mineralização e a expressão de proteínas ósseas.
OBJECTIVE: To examine if triiodothyronine (T3) increases osteogenic differentiation adipose tissue derived stem cells (ASCs) from ovariectomized adult rats with osteoporosis compared with young rats and adult rats without osteoporosis. MATERIALS AND METHODS: The ASCs were cultured in osteogenic medium and distributed into seven groups: 1) ASCs of young rats without osteoporosis; 2) ASCs of adult rats without osteoporosis; 3) ASCs of adult rats with osteoporosis and 4, 5, 6 and 7) ASCs of adult rats with osteoporosis treated with T3 (0.01 nM, 1 nM, 100 nM and 1,000 nM). We analyzed alkaline phosphatase activity, dimethylthiazol (MTT) conversion, percentage of mineralized nodules, cellularity and quantification of gene transcripts for collagen I, osteocalcin, osteopontin and Bmp-2. RESULTS: Regardless of the dose, T3 reduced the MTT conversion, alkaline phosphatase activity, percentage of cells and the expression of collagen I in at least one of the doses and periods studied (p < 0.05). But, the treatment with T3 does not modify the number of mineralized nodules and the expression of osteopontin and Bmp-2 in culture of ASCs from adult rats with osteoporosis (p > 0.05). CONCLUSION: T3 has a negative effect on some factors involved in osteogenic differentiation of ASCs from adult rats with osteoporosis, without; however, reduce the formation of mineralized nodules and the expression of bone proteins.
Assuntos
Animais , Feminino , Ratos , Tecido Adiposo/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteoporose , Osteogênese/efeitos dos fármacos , Tri-Iodotironina/farmacologia , Fatores Etários , Fosfatase Alcalina/metabolismo , Diferenciação Celular , Células-Tronco Mesenquimais/enzimologia , Células-Tronco Mesenquimais/fisiologia , Ovariectomia , Osteogênese/fisiologia , Osteoporose/patologia , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo RealRESUMO
OBJECTIVE: To examine if triiodothyronine (T3) increases osteogenic differentiation in bone marrow mesenchymal stem cells (BMMSCs) of adult rats compared with young rats. MATERIALS AND METHODS: BMMSCs were cultured in osteogenic medium and distributed into six groups: 1) BMMSCs of young rats; 2) BMMSCs of adult rats; 3, 4, 5 and 6) BMMSCs of adult rats with T3 (0.01, 1, 100 to 1000 nM). We analyzed alkaline phosphatase activity, dimethylthiazol (MTT) conversion, and collagen synthesis at 7, 14, and 21 days, and percentage of cells per field and number of mineralized nodules at 21 days of differentiation. RESULTS: T3 reduced MTT conversion, alkaline phosphatase activity, collagen synthesis, and the synthesis of mineralizalized nodules in at least one of the doses and periods studied (p < 0.05). Values were lower when compared with young and adult rats BMMSCs (p < 0.05) without T3. CONCLUSION: T3 has a negative effect on the factors involved in osteogenic differentiation of BMMSC from adult rats.
Assuntos
Células da Medula Óssea/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Tri-Iodotironina/farmacologia , Fosfatase Alcalina/metabolismo , Análise de Variância , Animais , Células da Medula Óssea/citologia , Calcificação Fisiológica/efeitos dos fármacos , Células Cultivadas , Colágeno/metabolismo , Feminino , Células-Tronco Mesenquimais/citologia , Modelos Animais , Fenótipo , Ratos , Ratos Wistar , Sais de Tetrazólio/metabolismo , Tiazóis/metabolismoRESUMO
OBJETIVO: Avaliar se a adição de T3 aumenta o potencial osteogênico das células-tronco mesenquimais da medula óssea (CTM-MO) de ratas adultas normais comparado ao de ratas jovens. MATERIAIS E MÉTODOS: CTM-MO foram cultivadas em meio osteogênico e separadas em seis grupos: 1) CTM-MO de ratas jovens; 2) CTM-MO de ratas adultas; 3, 4, 5 e 6) CTM-MO de ratas adultas com T3 nas concentrações de 0,01; 1; 100 e 1000 nM, respectivamente. Foram avaliados: atividade da fosfatase alcalina, conversão do dimetiltiazol (MTT) e síntese de colágeno aos sete, 14 e 21 dias e celularidade e número de nódulos de mineralização aos 21 dias de diferenciação. RESULTADOS: T3 reduziu significativamente a conversão do MTT, a atividade da fosfatase alcalina, a síntese de colágeno e a formação dos nódulos de mineralização em pelo menos uma das doses e dos períodos estudados (p < 0,05). Os valores foram menores quando comparados aos das CTM-MO de ratas jovens e adultas sem T3 (p < 0,05). CONCLUSÃO: T3 apresenta efeitos negativos sobre os fatores envolvidos na diferenciação osteogênica das CTM-MO de ratas adultas.
OBJECTIVE: To examine if triiodothyronine (T3) increases osteogenic differentiation in bone marrow mesenchymal stem cells (BMMSCs) of adult rats compared with young rats. MATERIALS AND METHODS: BMMSCs were cultured in osteogenic medium and distributed into six groups: 1) BMMSCs of young rats; 2) BMMSCs of adult rats; 3, 4, 5 and 6) BMMSCs of adult rats with T3 (0.01, 1, 100 to 1000 nM). We analyzed alkaline phosphatase activity, dimethylthiazol (MTT) conversion, and collagen synthesis at 7, 14, and 21 days, and percentage of cells per field and number of mineralized nodules at 21 days of differentiation. RESULTS: T3 reduced MTT conversion, alkaline phosphatase activity, collagen synthesis, and the synthesis of mineralizalized nodules in at least one of the doses and periods studied (p < 0.05). Values were lower when compared with young and adult rats BMMSCs (p < 0.05) without T3. CONCLUSION: T3 has a negative effect on the factors involved in osteogenic differentiation of BMMSC from adult rats.
Assuntos
Animais , Feminino , Ratos , Células da Medula Óssea/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Tri-Iodotironina/farmacologia , Análise de Variância , Fosfatase Alcalina/metabolismo , Células da Medula Óssea/citologia , Células Cultivadas , Calcificação Fisiológica/efeitos dos fármacos , Colágeno/metabolismo , Modelos Animais , Células-Tronco Mesenquimais/citologia , Fenótipo , Ratos Wistar , Sais de Tetrazólio/metabolismo , Tiazóis/metabolismoRESUMO
A rare case of anomalous chordae tendineae associated with mitral valve dysplasia was observed during necropsy in a 5- year-old male Persian cat with a history of sudden death. Grossly, there was thickening of the mitral valve, which was connected to numerous, short and thickened chordae tendineae that were ectopically inserted into the myocardium. In addition, marked left atrial dilation and pulmonary congestion and edema were present.
Assuntos
Animais , Cordas Tendinosas , Doenças das Valvas Cardíacas/veterinária , Doenças do Gato , Valva Mitral/anormalidadesRESUMO
A rare case of anomalous chordae tendineae associated with mitral valve dysplasia was observed during necropsy in a 5- year-old male Persian cat with a history of sudden death. Grossly, there was thickening of the mitral valve, which was connected to numerous, short and thickened chordae tendineae that were ectopically inserted into the myocardium. In addition, marked left atrial dilation and pulmonary congestion and edema were present.(AU)