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1.
Tissue Eng Part A ; 26(13-14): 769-779, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32493133

RESUMO

There is a constant need for improving embryo culture conditions in assisted reproduction. One possibility is to use mesenchymal stem/stromal cells derived from menstrual blood (mbMSCs), with an endometrial origin. In this study, we sought to analyze the expansion of mouse embryos in a direct coculture model with mbMSCs. Our results showed that after five passages, mbMSCs presented a spindle-shaped morphology, with surface markers that were comparable with the normal mesenchymal cell phenotype. mbMSCs could differentiate into adipogenic and osteogenic lineages and secrete angiopoetin-2 and hepatocyte growth factor. The coculture experiments employed 103 two-cell-stage embryos that were randomly divided into two groups: control (n = 50), embryos cultured in GV-Blast medium, and cocultured mbMSCs (n = 53), embryos cocultured with GV-Blast and mbMSCs. Typically, two to three embryos were placed in a well with 200 µL of culture medium and observed until developmental day 5. After 5 days, the cocultured group had more embryos in the blastocyst stage (69.8%) when compared with the control group (30%) (p < 0.001). It was also found that nearly 57% of blastocysts in the cocultured group reached the hatching stage, while only 13% achieved this stage in the control group (p < 0.001). Analyses of cultured mbMSCs and growth media, in the presence or absence of an embryo, were also performed. Immunofluorescence detected similar levels of collagen I and III and fibronectin in both mbMSCs and cocultured mbMSCs, and similar amounts of growth factors, VEGF, PDGF-AA, and PDGF-BB, were also observed in the conditioned medium, regardless of embryo presence. The present study describes, for the first time, an easy, noninvasive, and autologous method that could potentially increase blastocyst growth rates during assisted reproductive procedures (i.e., in vitro fertilization). It is proposed that this mbMSC coculture strategy enriches the embryonic microenvironment and promotes embryo development. This technique may complement or replace existing assisted reproduction methods and is directly relevant to the field of personalized medicine. Impact statement The study demonstrates a novel and potentially personalized assisted reproduction approach. The search for alternative and autologous methods provides assisted reproduction patients with a better chance of a successful pregnancy. In this study, mesenchymal cells derived from menstrual blood resembled the outside uterine surface and could potentially be employed for improving embryo outgrowth. Our protocol enriches the embryonic microenvironment and facilitates high-quality single-embryo transfer.


Assuntos
Desenvolvimento Embrionário/fisiologia , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Angiopoietina-2/metabolismo , Blastocisto/citologia , Blastocisto/metabolismo , Diferenciação Celular/fisiologia , Células Cultivadas , Meios de Cultivo Condicionados , Embrião de Mamíferos/citologia , Embrião de Mamíferos/metabolismo , Desenvolvimento Embrionário/genética , Endométrio/citologia , Endométrio/metabolismo , Feminino , Fibronectinas/metabolismo , Fator de Crescimento de Hepatócito/metabolismo , Humanos
2.
Sci Rep ; 8(1): 14751, 2018 10 03.
Artigo em Inglês | MEDLINE | ID: mdl-30282988

RESUMO

Maternal diet plays a critical role in health development. Perinatal overnutrition induces metabolic dysfunctions and obesity in the offspring. Obesity is associated with endocannabinoid system (ECS) over activation and oxidative stress. Liver ECS activation induces hepatic steatosis, inflammation and fibrosis while the antagonism of cannabinoid receptors ameliorates these alterations. Here, we investigated the effect of perinatal maternal high-fat diet (HF, 29% of calories as fat) on the ECS and antioxidant system in liver of male and female adult rat offspring (180 days old). Maternal HF diet increased hepatic cannabinoid receptors, ECS metabolizing enzymes and triglyceride content, with male offspring more affected. ECS changes are likely independent of estradiol serum levels but associated with increased hepatic content of estrogen receptor, which can stimulate the expression of ECS components. Differently, maternal HF diet decreased the activity of the antioxidant enzymes glutathione peroxidase, superoxide dismutase and catalase, and increased oxidative stress markers in both sexes. Alterations in the redox homeostasis were associated with mitochondria damage but not with liver fibrosis. Our data suggest that maternal HF diet induces ECS over activation in adulthood, and that male offspring are at higher risk to develop liver disease compared with female rats.


Assuntos
Dieta Hiperlipídica/efeitos adversos , Fígado Gorduroso/genética , Cirrose Hepática/genética , Fígado/metabolismo , Obesidade/genética , Efeitos Tardios da Exposição Pré-Natal/genética , Receptores de Canabinoides/genética , Animais , Catalase/genética , Catalase/metabolismo , Endocanabinoides/metabolismo , Estradiol/sangue , Fígado Gorduroso/etiologia , Fígado Gorduroso/metabolismo , Fígado Gorduroso/patologia , Feminino , Regulação da Expressão Gênica , Glutationa Peroxidase/genética , Glutationa Peroxidase/metabolismo , Homeostase/genética , Fígado/patologia , Cirrose Hepática/etiologia , Cirrose Hepática/metabolismo , Cirrose Hepática/patologia , Masculino , Obesidade/etiologia , Obesidade/metabolismo , Obesidade/patologia , Oxirredução , Estresse Oxidativo , Gravidez , Efeitos Tardios da Exposição Pré-Natal/etiologia , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Efeitos Tardios da Exposição Pré-Natal/patologia , Fenômenos Fisiológicos da Nutrição Pré-Natal , Ratos , Receptores de Canabinoides/metabolismo , Receptores de Estrogênio/genética , Receptores de Estrogênio/metabolismo , Fatores Sexuais , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo , Triglicerídeos/metabolismo
3.
Rio de Janeiro; s.n; 2013. 67 p. ilus.
Tese em Português | LILACS | ID: lil-719623

RESUMO

A Organização Mundial de Saúde (OMS) estima para 2030, 27 milhões de casos incidentes de câncer. No Brasil, segundo o Instituto Nacional do Câncer, foram estimados 518.510 casos novos de câncer para os anos de 2012 e 2013. Dessa estimativa, 52.680 correspondem ao câncer de mama (CM), com um risco estimado de 52 novos casos a cada 100.000 mulheres. O câncer de mama é um dos tipos de câncer mais comuns no mundo todo. Hoje se sabe que o tratamento para o CM pode levar ao surgimento de diferentes efeitos adversos tardios, entre eles a osteoporose. Uma das principais causas de surgimento da osteoporose é a menopausa precoce, que ocorre através da diminuição da concentração de estrogênio sérico. Este trabalho teve como objetivo avaliar os efeitos na matriz óssea induzidos pela quimioterapia, simulando um tratamento para o CM, em ratas Wistar.Ratas Wistar, com aproximadamente 3 meses de idade, foram divididas em: grupo controle e grupo que recebeu quimioterapia com poliquimioterápico docetaxel + ciclofosfamida (TC). A quimioterapia foi administrada em 4 ciclos, com intervalo de 1 semana entre eles. Os ratos foram submetidos à eutanásia 5 meses após o término do tratamento, para que os efeitos tardios pudessem ser avaliados. Vários estudos foram conduzidos: dosagem sorológica de estradiol, ensaios histológicos através de imunohistoquímica, micro-fluorescência de Raios-X, micro-tomografia computadorizada. Além de microscopia eletrônica de transmissão e varredura.Analisando os resultados obtidos em conjunto, sugere-se que a etapa inicial para o desenvolvimento da osteoporose, causada pelo poliquimioterápico TC, seja a diminuição da função ovariana. Este evento leva à diminuição da concentração de estrogênio sérico, o que causa a atrofia uterina. Concomitante a estes fatos, o TC causa redução na concentração de zinco no tecido ósseo. Estes resultados associados causam um desequilíbrio na relação osteoblastos/osteoclastos no osso...


The World Health Organization (WHO) estimates for 2030, 27 million incident cases of cancer. In Brazil, according to the National Cancer Institute, there were estimated 518,510 new cases of cancer for the years 2012 and 2013. From this estimation, 52,680 will correspond to breast cancer (BC), with an estimated risk of 52 new cases per 100,000 women. BC is one of the most common cancers worldwide. Today it is known that the treatment of the BC may lead to the emergence of different late adverse effects, including osteoporosis. One of the main causes of osteoporosis is the early menopause, which occurs with decreasing the serum estrogen concentration. This study aimed to evaluate the effects on bone matrix induced by chemotherapy , simulating a treatment for BC in Wistar rats .Wistar rats, approximately 3 months old, were divided into a control group and the group receiving polichemotherapy with docetaxel + cyclophosphamide (TC). Chemotherapy was administered in 4 cycles, with an interval of 1 week between them. The rats were euthanized 5 months after the end of treatment, so that the late effects could be evaluated. Several studies were performed: dosage of serum estradiol levels, histological tests by immunohistochemistry, micro X-ray fluorescence, micro-computed tomography and also transmission and scanning electron microscopy.Analyzing the results together, it is suggested that the initial step in the development of osteoporosis caused by multidrug TC is the reduction in the ovarian function. This event leads to decreased serum concentration of estrogen, which causes uterine atrophy. Concomitant to these facts, the TC causes a reduction in the concentration of zinc in the bone tissue. These results associated cause an imbalance in the osteoblast/osteoclast ratio in the bone tissue. The reduction in estrogen leads to decreased apoptosis of osteoclasts, while the reduction of zinc inhibits osteoblast function. This imbalance affects the bone turnover...


Assuntos
Animais , Ratos , Protocolos de Quimioterapia Combinada Antineoplásica , Matriz Óssea , Neoplasias da Mama/tratamento farmacológico , Antineoplásicos/farmacologia , Ciclofosfamida/administração & dosagem , Ciclofosfamida/farmacologia , Densidade Óssea , Menopausa Precoce , Osteoporose/etiologia , Ratos Wistar , Taxoides/administração & dosagem , Taxoides/farmacologia
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