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1.
Acta Histochem ; 125(8): 152112, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37948785

RESUMO

Kisspeptin (Kp-10) is a neuropeptide that binds to GPR54 receptors, exerting several functions mainly in the nervous and reproductive systems of the body. However, its effects and mechanisms of action on the skeletal system remain poorly understood. This study evaluated the effects of different concentrations of Kp-10 on in vitro osteogenic differentiation of multipotent mesenchymal stromal cells (MSCs) extracted from the bone marrow (BM) of adult Wistar rats. Two-month-old female rats were euthanized to extract BM from long bones to obtain MSCs. Four experimental groups were established in vitro: a control and Kp-10 at concentrations of 0.01, 0.05 and, 0.1 µg/mL. After induction of osteogenic differentiation, cell viability was evaluated using the 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl tetrazolium bromide (MTT) assay, alkaline phosphatase activity, collagen synthesis, percentage of area covered by MSCs/field and mineralized nodules/field, and immunocytochemistry of the GPR54 receptor tests. Furthermore, evaluation of gene transcripts for type I collagen, Runx-2, Bmp-2, bone sialoprotein, osteocalcin and osteopontin was performed using real-time RT-qPCR. It was observed that MSCs expressed GPR54 receptor to which Kp-10 binds during osteogenic differentiation, promoting a negative effect on osteogenic differentiation. This effect was observed at all the Kp-10 concentrations in a concentration-dependent manner, characterized by a decrease in the activity of alkaline phosphatase, collagen synthesis, mineralized nodules, and decreased expression of gene transcripts for type I collagen, osteocalcin, osteopontin, and Runx-2. Thus, Kp-10 inhibits in vitro osteogenic differentiation of MSCs extracted from the BM of adult Wistar rats.


Assuntos
Kisspeptinas , Células-Tronco Mesenquimais , Osteogênese , Animais , Feminino , Ratos , Fosfatase Alcalina/metabolismo , Medula Óssea/metabolismo , Células da Medula Óssea/metabolismo , Diferenciação Celular , Células Cultivadas , Colágeno Tipo I/metabolismo , Kisspeptinas/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Osteocalcina/genética , Osteogênese/efeitos dos fármacos , Osteogênese/fisiologia , Osteopontina/metabolismo , Osteopontina/farmacologia , Ratos Wistar
2.
J Oral Pathol Med ; 51(2): 206-216, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-35048460

RESUMO

BACKGROUND: Central giant cell granulomas (CGCG) of the jaws are osteolytic lesions that may behave aggressively and respond poorly to surgery. Microscopically, in addition to giant cells, there is a mononuclear cell population composed of macrophage/monocytic cells and spindle-shaped cells of mesenchymal origin. Seventy two percent of these tumours harbour mutually exclusive TRPV4, KRAS and FGFR1 mutations. We aimed to assess the mutational status of mononuclear and giant cells and the osteogenic potential of stromal cells in vitro and in vivo. METHODS AND RESULTS: We screened CGCG for signature mutations and used laser-capture microdissection to demonstrate that the mutations are restricted to the mononuclear cells. Additionally, we established CGCG primary cell culture and observed that the cells retained the mutations throughout passages. By flow cytometry, we observed predominance of CD14- CD51- CD61- cells, consistent with the expected profile for stromal cells. Considering the mesenchymal origin of stromal cells, we assessed the osteogenic differentiation potential of CGCG cells in culture by cytochemistry (von Kossa and alizarin red staining), alkaline phosphatase (ALP) activity assay and gene expression of osteogenic markers. CGCG cells presented self-capacity to increase ALP levels in a time-dependent manner and under osteogenic induction presented increasing number of calcium deposits, and overall higher expression of osteocalcin, RUNX2, ALPL and osteopontin than cells without osteogenic induction. A patient-derived xenograft model for CGCG was established, and osteoid material deposition was observed. CONCLUSION: Collectively, the results confirm that the signature mutations are restricted to stromal cells in CGCG, and the in vitro and in vivo results support that these cells have the capacity to differentiate into osteoblasts, in line with the bone formation often observed in the stroma of these lesions.


Assuntos
Granuloma de Células Gigantes , Células-Tronco Mesenquimais , Fosfatase Alcalina , Diferenciação Celular , Células Cultivadas , Granuloma de Células Gigantes/genética , Humanos , Arcada Osseodentária , Mutação , Osteogênese/genética , Células Estromais
3.
Cartilage ; 13(2_suppl): 839S-846S, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-31441318

RESUMO

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 , Ratos
4.
J Anim Physiol Anim Nutr (Berl) ; 104(5): 1256-1266, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32281708

RESUMO

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 , Ratos
5.
BMC Genomics ; 20(1): 866, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31730444

RESUMO

BACKGROUND: The white-eared opossum (Didelphis albiventris) is widely distributed throughout Brazil and South America. It has been used as an animal model for studying different scientific questions ranging from the restoration of degraded green areas to medical aspects of Chagas disease, leishmaniasis and resistance against snake venom. As a marsupial, D. albiventris can also contribute to the understanding of the molecular mechanisms that govern the different stages of organogenesis. Opossum joeys are born after only 13 days, and the final stages of organogenesis occur when the neonates are inside the pouch, depending on lactation. As neither the genome of this opossum species nor its transcriptome has been completely sequenced, the use of D. albiventris as an animal model is limited. In this work, we sequenced the D. albiventris transcriptome by RNA-seq to obtain the first catalogue of differentially expressed (DE) genes and gene ontology (GO) annotations during the neonatal stages of marsupial development. RESULTS: The D. albiventris transcriptome was obtained from whole neonates harvested at birth (P0), at 5 days of age (P5) and at 10 days of age (P10). The de novo assembly of these transcripts generated 85,338 transcripts. Approximately 30% of these transcripts could be mapped against the amino acid sequences of M. domestica, the evolutionarily closest relative of D. albiventris to be sequenced thus far. Among the expressed transcripts, 2077 were found to be DE between P0 and P5, 13,780 between P0 and P10, and 1453 between P5 and P10. The enriched GO terms were mainly related to the immune system, blood tissue development and differentiation, vision, hearing, digestion, the CNS and limb development. CONCLUSIONS: The elucidation of opossum transcriptomes provides an out-group for better understanding the distinct characteristics associated with the evolution of mammalian species. This study provides the first transcriptome sequences and catalogue of genes for a marsupial species at different neonatal stages, allowing the study of the mechanisms involved in organogenesis.


Assuntos
Sequenciamento do Exoma/estatística & dados numéricos , Regulação da Expressão Gênica no Desenvolvimento , Gambás/genética , Proteínas/genética , Transcriptoma , Animais , Animais Recém-Nascidos , Brasil , Ontologia Genética , Anotação de Sequência Molecular , Gambás/crescimento & desenvolvimento , Gambás/metabolismo , Proteínas/classificação , Proteínas/metabolismo , Análise de Sequência de RNA
6.
Acta Histochem ; 120(6): 505-512, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29907324

RESUMO

OBJECTIVE: Evaluate the effects of doses of caffeine administered to pregnant rats on the articular cartilage chondrocytes of their offspring. METHODS: Twenty-four adult Wistar rats were randomly assigned to four groups, with one control group and three groups being treated with caffeine at doses of 25, 50 and 100 mg/kg throughout pregnancy. At birth, three offspring/females were euthanized so that the chondrocytes could be extracted. At 7, 14 and 21 days of culture, the chondrocytes were subjected to the MTT cell viability assay and an evaluation of their alkaline phosphatase activity and collagen synthesis. Chondrocytes were also stained by Hematoxylin-eosin, PAS, Safranin-O and Alcian Blue. The Sox-9, Runx-2, aggrecan, collagen-II and alkaline phosphatase gene transcript levels were also evaluated. Mean comparisons were performed by the Student-Newman-Keuls test. RESULTS: Chondrocyte cultures from the 25 mg/kg group had the lowest results, as chondrocytes from this group had reduced viability, percentage of cells, alkaline phosphatase activity and collagen and chondrogenic matrix synthesis. A reduced expression of Sox-9, alkaline phosphatase and collagen-II was also detected in the 25 mg/kg group. Chondrocyte cultures of the group treated with 50 mg/kg caffeine showed reduced collagen synthesis and Sox-9 expression. The caffeine dose of 100 mg/kg also reduced collagen and Sox-9 and alkaline phosphatase expression. CONCLUSION: Caffeine administered to pregnant rats negatively alters the articular cartilage chondrocytes of their offspring, reducing the synthesis of collagen and Sox-9 expression regardless of the dose. This study also concluded that the effects of caffeine are not linear or dose-dependent.


Assuntos
Cafeína/efeitos adversos , Cartilagem Articular/metabolismo , Condrócitos/metabolismo , Colágeno/biossíntese , Regulação da Expressão Gênica/efeitos dos fármacos , Exposição Materna/efeitos adversos , Animais , Animais Recém-Nascidos , Cafeína/farmacologia , Cartilagem Articular/patologia , Sobrevivência Celular/efeitos dos fármacos , Condrócitos/patologia , Feminino , Gravidez , Ratos , Ratos Wistar , Fatores de Transcrição SOX9/metabolismo
7.
Arch Endocrinol Metab ; 60(2): 130-7, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27191047

RESUMO

Objective To evaluate, in rat offspring, bone changes induced by excess maternal thyroxin during pregnancy and lactation, and to assess the reversibility of these changes after weaning. Material and methods Twenty Wistar rats were distributed in two groups, hyperthyroid and control, that were treated daily with L-thyroxin (50 mcg/animal) and placebo, respectively. The treatment was initiated seven days before mating and continued throughout pregnancy and lactation. From every female of each of the two groups, two offspring were euthanized after birth, two at 21 days of age (weaning), and two at 42 days of age (21 days after weaning). In newborns, the length of pelvic and thoracic limbs were measured, and in the other animals, the length and width of the femur and humerus were measured. Bones were dissected, decalcified, embedded in paraffin, and analyzed histomorphometrically. Results Excess maternal thyroxin significantly reduced the length of the pelvic limb in neonates. In 21-day-old individuals, excess maternal thyroxine reduced the length and the width of the femur and the humerus. It also increased thickness of the epiphyseal plate and the percentage of trabecular bone tissue. In 42-day-old individuals, there were no significant differences between groups in relation to the parameters evaluated in the previous periods. Conclusion Excess maternal thyroxine reduced growth in suckling rats both at birth and at weaning, and it also increased the percentage of trabecular bone tissue in 21-day-old animals. These changes, however, were reversible at 42 days, i.e., 21 days after weaning. Arch Endocrinol Metab. 2016;60(2):130-7.


Assuntos
Osso e Ossos/efeitos dos fármacos , Osso e Ossos/patologia , Troca Materno-Fetal , Tiroxina/farmacologia , Fatores Etários , Animais , Animais Recém-Nascidos/crescimento & desenvolvimento , Osso e Ossos/metabolismo , Feminino , Lactação/efeitos dos fármacos , Masculino , Gravidez , Ratos Wistar , Tiroxina/metabolismo , Fatores de Tempo , Desmame
8.
Arch. endocrinol. metab. (Online) ; 60(2): 130-137, Apr. 2016. tab, graf
Artigo em Inglês | LILACS | ID: lil-782160

RESUMO

ABSTRACT Objective To evaluate, in rat offspring, bone changes induced by excess maternal thyroxin during pregnancy and lactation, and to assess the reversibility of these changes after weaning. Material and methods Twenty Wistar rats were distributed in two groups, hyperthyroid and control, that were treated daily with L-thyroxin (50 mcg/animal) and placebo, respectively. The treatment was initiated seven days before mating and continued throughout pregnancy and lactation. From every female of each of the two groups, two offspring were euthanized after birth, two at 21 days of age (weaning), and two at 42 days of age (21 days after weaning). In newborns, the length of pelvic and thoracic limbs were measured, and in the other animals, the length and width of the femur and humerus were measured. Bones were dissected, decalcified, embedded in paraffin, and analyzed histomorphometrically. Results Excess maternal thyroxin significantly reduced the length of the pelvic limb in neonates. In 21-day-old individuals, excess maternal thyroxine reduced the length and the width of the femur and the humerus. It also increased thickness of the epiphyseal plate and the percentage of trabecular bone tissue. In 42-day-old individuals, there were no significant differences between groups in relation to the parameters evaluated in the previous periods. Conclusion Excess maternal thyroxine reduced growth in suckling rats both at birth and at weaning, and it also increased the percentage of trabecular bone tissue in 21-day-old animals. These changes, however, were reversible at 42 days, i.e., 21 days after weaning. Arch Endocrinol Metab. 2016;60(2):130-7.


Assuntos
Animais , Masculino , Feminino , Gravidez , Tiroxina/farmacologia , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/patologia , Troca Materno-Fetal , Tiroxina/metabolismo , Fatores de Tempo , Desmame , Osso e Ossos/metabolismo , Lactação/efeitos dos fármacos , Fatores Etários , Ratos Wistar , Animais Recém-Nascidos/crescimento & desenvolvimento
9.
Connect Tissue Res ; 57(2): 131-42, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26634797

RESUMO

Caffeine is an alkaloid that is widely consumed due to its presence in drugs, coffee, tea, and chocolate. This compound passes to offspring through the placenta and milk; can cause teratogenic mutations; and reduces the formation, growth, and mass of bone. Because mesenchymal stem cells (MSCs) are responsible for generating the entire skeleton, we hypothesized that these cells are targets of caffeine. This study evaluated the osteogenic differentiation of MSCs derived from the offspring of rats treated with caffeine during pregnancy and lactation. Twenty-four adult Wistar rats were randomly divided into four groups, including one control group and three experimental groups treated with 25, 50, or 100 mg/kg of caffeine. At weaning, three 21-day-old pups from each dam in each group were euthanized for extraction of bone marrow cells for in vitro tests. Caffeine doses of 50 and 100 mg/kg significantly reduced the activity of alkaline phosphatase at 7, 14, and 21 days and the expression of collagen I at 21 days. However, the expression of gene transcripts for alkaline phosphatase, Runx-2, and bone sialoprotein, as well as the synthesis of mineralization nodules, decreased significantly in all groups treated with caffeine. The expression of osteocalcin was significantly reduced only in the group treated with 50 mg/kg caffeine. The caffeine that passes from the mother to the offspring during pregnancy and lactation reduces the osteogenic differentiation of MSCs. We propose that this reduction in the osteogenic potential of MSCs may be involved in the pathogenesis of osteopenia resulting from caffeine consumption.


Assuntos
Cafeína/farmacologia , Diferenciação Celular/efeitos dos fármacos , Lactação/efeitos dos fármacos , Células-Tronco Mesenquimais/citologia , Osteogênese/efeitos dos fármacos , Fosfatase Alcalina/metabolismo , Animais , Antígenos CD/metabolismo , Calcificação Fisiológica/efeitos dos fármacos , Células Cultivadas , Feminino , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/enzimologia , Células-Tronco Mesenquimais/metabolismo , Fenótipo , Gravidez , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real
10.
BMC Musculoskelet Disord ; 16: 10, 2015 Feb 04.
Artigo em Inglês | MEDLINE | ID: mdl-25649420

RESUMO

BACKGROUND: Caffeine is an active alkaloid that can cause damage to bones in formation during prenatal life into adulthood. This compound can pass across the placenta and into the mother's milk, causing a reduction in bone formation, growth and mass. The objective of this study was to examine the osteogenic potential of osteoblasts extracted from neonatal rats born to mothers treated with caffeine throughout pregnancy. METHODS: Twenty-four adult Wistar rats were randomly divided into four groups, consisting of one control group and three groups that were treated with 25, 50, or 100 mg/kg of caffeine by an oral-gastric probe throughout the duration of the experimental period (pregnancy). At birth, three puppies from each dam in each group were euthanized, and osteoblasts were extracted from the calvaria of these pups for in vitro testing. RESULTS: The osteoblasts extracted from the pups of rats that received 50 mg/kg caffeine during pregnancy exhibited increased expression of osteocalcin, osteopontin, sialoprotein, runx-2, alkaline phosphatase and type I collagen transcripts, resulting in increased synthesis of mineralization nodules. CONCLUSIONS: Neonates from rats treated with 50 mg/kg caffeine during pregnancy contained osteoblasts with a higher osteogenic potential characterized by increased expression of osteocalcin, osteopontin, sialoprotein, runx-2, alkaline phosphatase and type I collagen and increased synthesis of mineralization nodules.


Assuntos
Cafeína/farmacologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/fisiologia , Osteogênese/efeitos dos fármacos , Prenhez/fisiologia , Fosfatase Alcalina/metabolismo , Animais , Cafeína/administração & dosagem , Calcificação Fisiológica , Diferenciação Celular , Sobrevivência Celular , Células Cultivadas , Colágeno/biossíntese , Cristalização , Feminino , Formazans/metabolismo , Expressão Gênica , Osteogênese/genética , Gravidez , Distribuição Aleatória , Ratos Wistar , Sais de Tetrazólio/metabolismo
11.
Ciênc. rural ; 43(12): 2234-2239, dez. 2013. ilus, tab
Artigo em Português | LILACS | ID: lil-691332

RESUMO

Foram avaliadas as propriedades mecânicas, força, deformação e tensão ao limite elástico, tensão ao ponto de ruptura e índice de rigidez, de meniscos frescos e meniscos transplantados por 70 dias em joelhos de coelhos, após preservação em glicerina. O primeiro grupo (tratado) foi formado por seis meniscos mediais do joelho esquerdo, coletados de animais oriundos de criação comercial. Esses meniscos foram armazenados em glicerina 98% por um período de 45-60 dias; em seguida, foram reidratados em solução salina a 0,5% de enrofloxacina, por 12 horas, e implantados em joelhos de outros seis coelhos, submetidos à meniscectomia. Após 70 dias da cirurgia, foi realizada a eutanásia dos animais e retirada dos aloenxertos para avaliação mecânica. O segundo grupo (controle) foi composto por seis meniscos mediais, retirados dos joelhos contralaterais (direito) dos mesmos animais. Para a amostra estudada, não houve diferença estatisticamente significante entre os grupos controle e tratado para nenhuma das variáveis analisadas, demonstrando que a reidratação de aloenxertos meniscais, após preservação em glicerina, mantém as características mecânicas desses, após 70 dias de implantação, semelhantes às de meniscos frescos.


We evaluated the mechanical properties, strength, strain and stress to the elastic limit, the rupture stress point and stiffness index of fresh menisci and menisci transplanted for 70 days in the knees of rabbits after preservation in glycerin. The first group (treated) was formed by six medial menisci of the left knee, collected from these animals for commercial breeding. These menisci were stored in 98% glycerin for a period of 30-60 days, and then rehydrated in a solution in 0.5% saline enrofloxacin for 12 hours, and implanted in the knees of another six rabbits underwent meniscectomy . After 70 days of surgery was performed the euthanasia of animals and removal of allografts for mechanical evaluation. The second group (control) consisted of six medial menisci, removed from the contralateral knees (right) of the same animals. For the sample studied, had no statistically significant difference between control and treated groups for any of the variables analyzed, showing that the rehydration of meniscal allografts after preservation in glycerin keeps the mechanical characteristics of these, after 70 days of implantation, similar to the meniscus fresh.

12.
Acta Ortop Bras ; 21(6): 307-9, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24453686

RESUMO

OBJECTIVE: To evaluate the in vitro effetcs of caffeine on proliferation, apoptosis a nd gene transcripts expression of chondrogenic differentiation in growth cartilage. METHODS: THE CARTILAGINOUS EPIPHYSES OF FEMURS OF NEWBORN RATS, WHICH WERE DIVIDED INTO TWO SUBGROUPS: treated with caffeine and control group, both observed over the time periods of 0, 7, 14 and 21 days. The cartilaginous epiphyses of femurs of each subgroup and each time span were subjected to histomorphometric, immunohistochemical analysis, Tunel technique and RT-PCR in real time. RESULTS: The decrease in proliferative activity and the increase of apoptotic chondroblasts at 21 days were found regardless of the subgroup. However, the decrease in cell proliferation caused by caffeine was lower than in the control group and significantly increased the expression of gene transcripts for chondrogenic differentiation, represented by Sox-9 and Runx-2. However, the in vitro culture with caffeine revealed antagonistic effects: despite the positive effect on chondroblasts proliferation and differentiation, caffeine increased apoptosis, characterized by increased expression of caspase 3 and of the number of cells undergoing apoptosis (p<0.05). CONCLUSION: Caffeine presents antagonistic effects in vitro on growth cartilage, increasing the proliferation, differentiation and cell apoptosis. Experimental Study .

13.
Acta ortop. bras ; 21(6): 307-309, 2013. ilus, tab
Artigo em Português | LILACS | ID: lil-689700

RESUMO

OBJETIVO: Avaliar os efeitos in vitro da cafeína na proliferação, apoptose e expressão de transcriptos gênicos de diferenciação condrogênica na cartilagem de crescimento.MÉTODO: As epífises cartilaginosas de fêmures de ratos neonatos foram divididas em dois subgrupos: os tratados com cafeína e o grupo controle, ambos observados nos tempos de 0, 7, 14 e 21 dias. As epífises cartilaginosas de fêmures de cada subgrupo e de cada tempo foram submetidas à histomorfometria, análise imunoistoquímica, técnica de túnel e RT-PCR em tempo real.RESULTADO: A diminuição da atividade proliferativa e o aumento de condroblastos em apoptose aos 21 dias foram encontrados em ambos os subgrupos. Entretanto a diminuição da proliferação celular causada pela cafeína foi menor quando comparada ao grupo controle e aumentou significativamente a expressão de transcriptos gênicos para diferenciação condrogênica, representada pelo SOX-9 e pelo RUNX-2. Entretanto o cultivo in vitro com cafeína demostrou efeitos antagônicos: apesar dos efeitos positivos na proliferação e diferenciação de condroblatos, cafeína aumentou a apoptose, caracterizada pelo aumento da expressão de caspase-3 e do numero de células em apoptose (p< 0.05).CONCLUSÃO: A cafeína apresenta efeitos antagônicos in vitro na cartilagem em crescimento, aumentando a proliferação, diferenciação e apoptose celular. Estudo experimental.


OBJECTIVE: To evaluate the in vitro effetcs of caffeine on proliferation, apoptosis and gene transcripts expression of chondrogenic differentiation in growth cartilage.METHODS: The cartilaginous epiphyses of femurs of newborn rats, which were divided into two subgroups: treated with caffeine and control group, both observed over the time periods of 0, 7, 14 and 21 days. The cartilaginous epiphyses of femurs of each subgroup and each time span were subjected to histomorphometric, immunohistochemical analysis, Tunel technique and RT-PCR in real time.RESULTS: The decrease in proliferative activity and the increase of apoptotic chondroblasts at 21 days were found regardless of the subgroup. However, the decrease in cell proliferation caused by caffeine was lower than in the control group and significantly increased the expression of gene transcripts for chondrogenic differentiation, represented by SOX-9 and RUNX-2. However, the in vitro culture with caffeine revealed antagonistic effects: despite the positive effect on chondroblasts proliferation and differentiation, caffeine increased apoptosis, characterized by increased expression of caspase 3 and of the number of cells undergoing apoptosis (p<0.05).CONCLUSION: Caffeine presents antagonistic effects in vitro on growth cartilage, increasing the proliferation, differentiation and cell apoptosis. Experimental Study.


Assuntos
Animais , Ratos , Apoptose , Diferenciação Celular , Cafeína/biossíntese , Cartilagem/crescimento & desenvolvimento , Epífises/crescimento & desenvolvimento , Fêmur , Proliferação de Células , Animais Recém-Nascidos , Ensaio Clínico , Ratos Wistar
14.
Ciênc. rural ; 40(5): 1114-1120, maio 2010. graf, ilus, tab
Artigo em Português | LILACS | ID: lil-552151

RESUMO

O presente estudo avaliou a resistência à compressão de meniscos mediais de coelhos da raça Nova Zelândia, por meio de teste mecânico de compressão. Trinta meniscos foram distribuídos em três grupos: grupo MF, composto por dez meniscos frescos; grupo MG, dez meniscos preservados em glicerina 98 por cento, por 30 dias, e grupo MR, dez meniscos preservados em glicerina 98 por cento, por 30 dias e reidratados em NaCl 0,9 por cento, por 12 horas. Os meniscos de cada grupo foram submetidos ao teste de compressão no sentido perpendicular ao seu plano anatômico regular e foram avaliados o limite de elasticidade, a deformação elástica, a tensão ao ponto de ruptura e ao limite de elasticidade e ainda, o índice de rigidez. Os meniscos dos grupos preservados, MG e MR, tiveram o limite elástico semelhante ao grupo de meniscos frescos (MF). O grupo de meniscos em glicerina (MG) apresentou menor capacidade de deformação elástica (P<0,05) que os grupos MF e MR, e maior capacidade de sofrer tensão ao limite elástico. Os meniscos do grupo (MG) apresentaram maior rigidez (P<0,05) que os meniscos dos grupos MF e MR. Conclui-se que o grupo de meniscos preservados em glicerina 98 por cento, seguido de reidratação em NaCl 0,9 por cento (MR), não apresentou alterações significativas na capacidade de resistência ao limite elástico dos meniscos.


The present study evaluated the compressive strength of medial menisci of New Zealand rabbits, through mechanical compression test. Thirty menisci were distributed in three groups: group MF, composed by ten fresh menisci; MG group, composed by ten menisci preserved in 98 percent glycerin for 30 days; and, group MR, ten menisci preserved in 98 percent glycerin for 30 days and rehydrated in NaCl 0.9 percent for 12 hours. The menisci in each group were submitted to compression test in the perpendicular direction to the anatomical plane and had the elasticity limit, the elastic deformity, the rupture stress point and the stiffness index evaluated. The menisci from the preserved groups MG and MR had the elastic limit similar to the fresh menisci group (MF). The group of menisci preserved in glycerin (MG) presented lower elastic deformity capacity (P<0.05) if compared to the other groups, MF and MR, and a higher tension capacity at elastic limit. The menisci from group (MG) presented higher stiffness (P<0.05) than the ones in the MF and MR groups. It can be concluded that the group menisci preserved in glycerin 98 percent followed by rehydration in Nacl 0,9 percent (MR), did not showed any significant alterations in the capacity of the menisci elastic limit.

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