Osteogenic differentiation of bone marrow mesenchymal stem cells of ovariectomized and non-ovariectomized female rats with thyroid dysfunction

Boeloni, Jankerle Neves; Ocarino, Natália de Melo; Silva, Juneo Freitas; Correa, Cristiane Rodrigues; Bertollo, Caryne Margotto; Hell, Rafaela Chitarra Rodrigues; Pereira, Marivalda M.; Góes, Alfredo M.; Serákides, Rogéria

doi:10.1016/j.prp.2012.10.004

Cited by 14 publications

(19 citation statements)

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“…Dessa forma, fica evidente que a idade do cão pode influenciar na capacidade de diferenciação das CTM-MO. Em ratos, as CTM-MO também tem seu potencial osteogênico comprometido pela idade (Hell et al 2012) e pela presença de doenças óssea ou endócrina (Boeloni et al 2013). A maior diferenciação osteogênica aliada à menor variação do potencial osteogê-nico, são algumas das razões, que justificam a utilização das CTM do tecido adiposo em defeitos ósseos.…”

Section: Discussionunclassified

“…Os cristais de formazan foram observados ao microscópio antes do acréscimo de 210mL de SDS (sódio dodecil sulfato)-10% HCl que permaneceu overnight em estufa a 37 o C e 5% de CO 2 . Posteriormente, 100mL de cada poço foram transferidos para placas de 96 poços para análise na leitora de placas com comprimento de onda de 595nm de acordo com Boeloni et al (2013).…”

Section: Comparação Do Potencial Osteogênico Das Ctm-ad E Ctm-mounclassified

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Estudo comparativo da diferenciação osteogênica das células tronco mesenquimais da medula óssea e do tecido adiposo de cães adultos

et al. 2016

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Resumo: O objetivo deste estudo foi comparar o potencial osteogênico das células tronco mesenquimais extraídas da medula óssea (CTM-MO) com as do tecido adiposo (CTM-AD) de cães adultos. As células foram caracterizadas fenotipicamente quanto à expressão de CD29, CD90, CD34 e CD45 e submetidas à diferenciação adipogênica e condrogênica por 21 dias e osteogênica por 7, 14 e 21 dias. Foram constituídos quatro grupos: 1) CTM-MO em meio osteogênico, 2) CTM-MO em meio basal, 3) CTM-AD em meio osteogênico e 4) CTM-AD em meio basal. Aos 7, 14 e 21 dias de diferenciação osteogênica as culturas foram submetidas às avaliações da conversão de MTT em formazan, da atividade da fosfatase alcalina (FA), da síntese de colágeno e de matriz mineralizada, avaliação do número de células por campo e foram quantificados os transcritos gênicos para osterix, sialoproteina óssea (BSP), osteonectina (ON) e osteocalcina (OC). Tanto as células extraídas da medula óssea quanto do tecido adiposo mostraram elevada expressão de marcadores para células tronco e baixa expressão de marcadores de células hematopoiéticas (menor que 2%). Além disso, foram capazes de se diferenciar em osteoblastos, condrócitos e adipócitos. As CTM-AD submetidas à diferenciação osteogênica mostraram maior conversão do MTT em formazan que as CTM-MO, sob mesmas condições aos 7 e 21 dias. O número de células por campo, a atividade da FA, a síntese de colágeno e de matriz mineralizada foram superior nas CTM-AD em diferenciação, em relação às CTM-MO sob as mesmas condições, em todos os tempos estudados. As expressões de osterix, BSP e OC foram predominantemente superiores nas CTM-MO diferenciadas, mas a expressão de ON foi superior nas CTM-AD diferenciadas aos 7, 14 e 21 dias. Conclui-se que as CTM-AD apresentam maior potencial osteogênico que as CTM-MO quando extraídas de cães adultos.

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Section: Discussionunclassified

Section: Comparação Do Potencial Osteogênico Das Ctm-ad E Ctm-mounclassified

Estudo comparativo da diferenciação osteogênica das células tronco mesenquimais da medula óssea e do tecido adiposo de cães adultos

et al. 2016

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“…At 0 and 19 days of gestation, blood was collected from the rats, and the serum was stored at -20°C for the measurement of free T4, which was performed by the chemiluminescence Elisa technique (sensitivity: 0.4 ng/dl), using commercial kits in accordance with the manufacturer’s instructions (IMMULITE Free T4, Siemens Medical Solutions Diagnostics, Malvern, PA, USA). This assay has been previously used and validated for rat [14,32,33]. The intra- and inter-assay coefficients of variation were 4% and 7%, respectively.…”

Section: Methodsmentioning

confidence: 99%

Luteal activity of pregnant rats with hypo-and hyperthyroidism

2014

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BackgroundLuteal activity is dependent on the interaction of various growth factors, cytokines and hormones, including the thyroid hormones, being that hypo- and hyperthyroidism alter the gestational period and are also a cause of miscarriage and stillbirth. Because of that, we evaluated the proliferation, apoptosis and expression of angiogenic factors and COX-2 in the corpus luteum of hypo- and hyperthyroid pregnant rats.MethodsSeventy-two adult female rats were equally distributed into three groups: hypothyroid, hyperthyroid and control. Hypo- and hyperthyroidism were induced by the daily administration of propylthiouracil and L-thyroxine, respectively. The administration began five days before becoming pregnant and the animals were sacrificed at days 10, 14, and 19 of gestation. We performed an immunohistochemical analysis to evaluate the expression of CDC-47, VEGF, Flk-1 (VEGF receptor) and COX-2. Apoptosis was evaluated by the TUNEL assay. We assessed the gene expression of VEGF, Flk-1, caspase 3, COX-2 and PGF2α receptor using real time RT-PCR. The data were analyzed by SNK test.ResultsHypothyroidism reduced COX-2 expression on day 10 and 19 (P < 0.05), endothelial/pericyte and luteal cell proliferation on day 10 and 14 (p < 0.05), apoptotic cell numbers on day 19 (p < 0.05) and the expression of Flk-1 and VEGF on day 14 and 19, respectively (p < 0.05). Hyperthyroidism increased the expression of COX-2 on day 19 (P < 0.05) and the proliferative activity of endothelial/pericytes cells on day 14 (p <0.05), as well as the expression of VEGF and Flk-1 on day 19 (P < 0.05).ConclusionsHypothyroidism reduces the proliferation, apoptosis and expression of angiogenic factors and COX-2in the corpus luteum of pregnant rats, contrary to what is observed in hyperthyroid animals, being this effect dependent of the gestational period.

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“…After four runs and until a cell confluence of 80-90% was obtained, the phenotypic characterization of the BMMSCs was performed by flow cytometry according to an established protocol. [33,39] The reading and analysis were performed on a Fluorescence Activated Cell Analyzer (FACScan) using Cell Quest software, with an acquisition of 20 000 events, with forward scatter (FSC) and side scatter (SSC) parameters on a linear scale and FL1 (relative fluorescence) on a logarithmic scale that detects light of 530 nm wavelength, which corresponds to green fluorescence, for analysis by the WinMDI program by dot plot charts and histograms. The primary antibodies used were as follows: anti-CD45 (clone 69 mouse), anti-CD54 (clone 1A29 mouse), anti-CD73 (clone 5 F/B9 mouse) and anti-CD90 (clone Ox-7 mouse) (BD Biosciences, San Jose, CA, USA).…”

Section: Phenotypic Characterizationmentioning

confidence: 99%

Dose-dependent effect of triiodothyronine on the chondrogenic differentiation of mesenchymal stem cells from the bone marrow of female rats

Assis

Elert

Azevedo

et al. 2017

Journal of Pharmacy and Pharmacology

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Objectives Verify the in-vitro effect of triiodothyronine (T3) on the chondrogenic differentiation of female rat bone marrow mesenchymal stem cells (BMMSCs) over several time periods and at several doses. Methods CD54 + /CD73 + /CD90 + BMMSCs from Wistar female rats were cultured in chondrogenic medium with or without T3 (0.01; 1; 100; 1000 nM). At seven, 14 and 21 days, the cell morphology, chondrogenic matrix formation and expression of Sox9 and collagen II were evaluated. Key findings The dose of 100 nM did not alter the parameters evaluated in any of the periods studied. However, the 0.01 nM T3 dose improved the chondrogenic potential by increasing the chondrogenic matrix formation and expression of Sox9 and collagen II in at least one of the evaluated periods; the 1 nM T3 dose also improved the chondrogenic potential by increasing the chondrogenic matrix formation and the expression of collagen II in at least one of the evaluated periods. The 1000 nM T3 dose improved the chondrogenic potential by increasing the chondrogenic matrix formation and Sox9 expression in at least one of the evaluated periods. Conclusions T3 has a dose-dependent effect on the differentiation of BMMSCs from female rats.

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Osteogenic differentiation of bone marrow mesenchymal stem cells of ovariectomized and non-ovariectomized female rats with thyroid dysfunction

Cited by 14 publications

References 50 publications

Estudo comparativo da diferenciação osteogênica das células tronco mesenquimais da medula óssea e do tecido adiposo de cães adultos

Estudo comparativo da diferenciação osteogênica das células tronco mesenquimais da medula óssea e do tecido adiposo de cães adultos

Luteal activity of pregnant rats with hypo-and hyperthyroidism

Dose-dependent effect of triiodothyronine on the chondrogenic differentiation of mesenchymal stem cells from the bone marrow of female rats

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