Utilização de células tronco mesenquimais autólogas para o tratamento de éguas com endometrite crônica degenerativa
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Data
2013-02-04
Autores
Pavão, Giovana D'Andrea [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
O trabalho teve por objetivo avaliar a eficiência da terapia celular no tratamento do endométrio de éguas com processo degenerativo crônico caracterizado pela presença de fibrose uterina. Foram utilizadas 10 éguas, de raça Quarto de Milha com idade entre 14 e 23 anos, com massa corpórea entre 400 a 600 Kg, e bom escore corporal com históricos reprodutivos de subfertilidade e grau severo de fibrose uterina detectada previamente pelo exame histológico realizado por meio da biópsia endometrial. O material foi coletada no dia zero (D0) antes do tratamento com células tronco mesenquimais (CTMs) e 15 dias (D15), 30 dias (D30) e 60 dias (D60) após o tratamento. As amostras foram classificadas segundo o modelo proposto por Kenney & Doig (1986) em grau de fibrose: Grau I - normal, Grau IIA – inflamação crônica leve, Grau IIB – inflamação crônica com infiltrado moderado, Grau III – inflamação crônica degenerativa grave. Estas amostras foram submetidas à confecção de lâminas para coloração de Tricrômio de Masson, Coloração de Hematoxilina e Eosina (HE) e Imunohistoquímica. A punção da medula óssea foi realizada em 10 éguas com idade entre 14 e 23 anos e o material coletado foi submetido ao Laboratório. O cultivo celular in vitro foi realizado em meio DMEM alta glicose em estufa com 37,5°C, em atmosfera úmida contendo 95% de ar e 5% de CO2. Após atingir a confluência de 80% na placa de cultivo, estas células foram removidas e injetadas intra-endometrial com uma concentração de 2x107 células\mL em um volume de 0,5 ml em 20 (vinte) diferentes pontos (sitio de administração) espaçados entre um centímetro, com o auxílio de endoscópio flexível, totalizando um volume de 10 mL aplicado. Nas avaliações realizadas, verificou-se diminuição significativa dos ninhos glandulares, mudança siginificativa de células...
The study aimed to evaluate the efficiency of cell therapy in the treatment of endometrial mares with chronic degenerative process characterized by the presence of uterine fibroids. We used 10 mares bred Quarter Horses aged between 14 and 23 years, with body mass between 400 and 600 kg, with good body and reproductive history of subfertility and severe degree of uterine fibrosis detected previously by histological examination performed by endometrial biopsy. The material was collected on day zero (D0) before treatment with mesenchymal stem cells (MSCs) and 15 days (D15), 30 days (D30) and day 60 (D60) post-treatment. The samples were classified according to the model proposed by Kenney & Doig (1986) in fibrosis: Grade I - normal, Grade IIA - mild chronic inflammation, IIB Grade - moderate chronic inflammatory infiltrate, Grade III - severe degenerative chronic inflammation. These samples were submitted for preparation of slides for staining Masson's trichrome, hematoxylin and eosin staining (HE) and immunohistochemistry. A bone marrow puncture was performed in 10 mares aged between 14 and 23 years and the collected material was submitted to the Laboratory. The in vitro cell cultivation was performed in DMEM with high glucose kiln 37.5 ° C in humidified atmosphere containing 95% air and 5% CO2. After reaching 80% confluence on plate culture, these cells were removed and injected intra-endometrial at a concentration of 2x107 cells \ ml in a volume of 0.5 ml for 20 (twenty) different point (site of administration) spaced of an inch, with the aid of flexible endoscope, for a total volume of 10 ml applied. In the assessments, there was a significant reduction in glandular nests, change polymorphonuclear cell siginificativa absent at baseline (D0) to moderate 60 days (D60); amendment of mononuclear cells in going from... (Complete abstract click electronic access below)
The study aimed to evaluate the efficiency of cell therapy in the treatment of endometrial mares with chronic degenerative process characterized by the presence of uterine fibroids. We used 10 mares bred Quarter Horses aged between 14 and 23 years, with body mass between 400 and 600 kg, with good body and reproductive history of subfertility and severe degree of uterine fibrosis detected previously by histological examination performed by endometrial biopsy. The material was collected on day zero (D0) before treatment with mesenchymal stem cells (MSCs) and 15 days (D15), 30 days (D30) and day 60 (D60) post-treatment. The samples were classified according to the model proposed by Kenney & Doig (1986) in fibrosis: Grade I - normal, Grade IIA - mild chronic inflammation, IIB Grade - moderate chronic inflammatory infiltrate, Grade III - severe degenerative chronic inflammation. These samples were submitted for preparation of slides for staining Masson's trichrome, hematoxylin and eosin staining (HE) and immunohistochemistry. A bone marrow puncture was performed in 10 mares aged between 14 and 23 years and the collected material was submitted to the Laboratory. The in vitro cell cultivation was performed in DMEM with high glucose kiln 37.5 ° C in humidified atmosphere containing 95% air and 5% CO2. After reaching 80% confluence on plate culture, these cells were removed and injected intra-endometrial at a concentration of 2x107 cells \ ml in a volume of 0.5 ml for 20 (twenty) different point (site of administration) spaced of an inch, with the aid of flexible endoscope, for a total volume of 10 ml applied. In the assessments, there was a significant reduction in glandular nests, change polymorphonuclear cell siginificativa absent at baseline (D0) to moderate 60 days (D60); amendment of mononuclear cells in going from... (Complete abstract click electronic access below)
Descrição
Palavras-chave
Tratamento veterinario, Células-Tronco, Egua, Endometrio, Stem cells, Endometrium
Como citar
PAVÃO, Giovana D'Andrea. Utilização de células tronco mesenquimais autólogas para o tratamento de éguas com endometrite crônica degenerativa. 2013. 66 f. Tese (doutorado) - Universidade Estadual Paulista Júlio de Mesquita Filho, Faculdade de Medicina Veterinária e Zootecnia de Botucatu, 2013.