Efeito da aplicação intratesticular de células tronco mesenquimais alogênicas em equinos
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Data
2018-01-31
Autores
Papa, Patricia de Mello
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
O presente estudo propõe avaliar a segurança da aplicação intratesticular de células tronco mesenquimais alogênicas obtidas da medula óssea e verificar o efeito DO tratamento com as células tronco mesenquimais (CTMs) em garanhões submetidos a degeneração testicular. Para o primeiro estudo, foram usados 24 garanhões sadios, nos quais os testiculos foram mensurados, termografia, ultrassonografia doppler, cinética espermática e dosagem hormonal. Os grupos foram separados aleatoriamente em grupo controle (GC) e grupo células tronco mesenquimais (CTMs). No GC foram realizadas aplicações intratesticulares com PBS (solução tampão) e no CTMs solução contendo células. Os animais foram avaliados após a aplicação e aos 15 dias foram orquiectomizados. Os fragmentos testiculares foram destinados para histologia. Os exames clínicos não mostraram declínio na condição geral dos animais após a aplicação e não houve diferença significativa nas variáveis volume, temperatura superficial e ultrassonografia testicular, cinética espermática, analise sérica hormonal e histologia em ambos os grupos. Desta forma, conclui-se que a aplicação intratesticular de CTMs alogênicas da medula óssea é um processo seguro, pois não provocou alterações locais. Para o segundo estudo, um grupo de 10 garanhões foram divididos em dois grupos: grupo controle (CT) e grupo tratado (TT). Os animais foram submetidos ao estresse térmico escrotal, acoplando uma bolsa termica nos testiculos, durante 3 horas por 3 dias consecutivos. Após uma semana da indução, os animais foram tratados com 10 x 106 CTMs suspensas em 5 ml de PBS em cada testiculo. O grupo controle recebeu o mesmo volume contendo apenas PBS. Biopsias testiculares foram coletadas em quatro diferentes momentos: antes do estresse termico (T0); depois do estresse termico (T1); após aplicação de PBS/CTMs (T2); e após 30 dias estresse termico (T3). O método usado para promover a degenerção testicular foi eficiente, mostrando alterações no epitélio seminófero dos dois grupos. Entretanto o grupo tratado com CTMs foi capaz de recuperar mais rápido os padrões iniciais da arquitetura testicular do que o grupo controle. Podemos concluir que a aplicação intratesticular com CTMs foi eficiente e promoveu uma recuperação rápida nos animais que sofreram degeneração testicular devido aos fatores regenerativos liberados pelas CTMs.
The present study aim to evaluate the safety of intratesticular application of allogenic mesenchymal stem cells from the bone marrow and verification through the histological examinations of the effect cells in animals induced to testicular degeneration. For the first study, a group of 24 healthy stallion, measurements of testicular volume, testicular surface temperature, Doppler ultrasonography, sperm kinetics and hormonal dosing were performed. The groups were randomly divided into a mesenchymal stem cell group (MSC) and control group (CG). In the first, an intratesticular application of CTM was performed and in the CG only a buffer solution (PBS) was used. After the application, the animals were examined again. Fifteen days later, the animals were orchiectomized and histological analysis of the fragments were performed. Clinical examination showed no decline in the overall condition of the animals after application. There was no difference in the analysed variables, such as testicular volume, testicular surface temperature, ultrassonography, sperm kinetics, hormonal analysis and histology, in both groups after application. Thus, the results of the presente study support the hypothesis of the feasibility and safety of intratesticular application of allogeneic bone marrow MSC.For the second study, a group of 10 stallions submitted to scrotal thermal stress, testicular histological samples were analyzed before and after thermal insult, after intratesticular applications (PBS / MSC) and after 30 days of insult. The treated and control groups, after thermal stress, received mesenchymal stem cells and PBS respectively in their testicles. The method used to promote degeneration was efficient showing alterations in the seminiferous epithelium of both groups, however the treated group was able to recover a histological preview after 30 days of insult, while it was not observed in the control group. The characteristics that MSC has to stimulate tissue angiogenesis and proliferation, release of growth factors, eliminate free radicals, increase the capacity for renewal and cell differentiation, its immunomodulatory and immunosuppressive actions may be responsible for the outcome in the present study.It is concluded that the application of intratesticular mesenchymal stem cells is safe, in addition to promoting restoration of the initial patterns of the histological architecture of the testis in animals induced to testicular degeneration by thermal stress.
The present study aim to evaluate the safety of intratesticular application of allogenic mesenchymal stem cells from the bone marrow and verification through the histological examinations of the effect cells in animals induced to testicular degeneration. For the first study, a group of 24 healthy stallion, measurements of testicular volume, testicular surface temperature, Doppler ultrasonography, sperm kinetics and hormonal dosing were performed. The groups were randomly divided into a mesenchymal stem cell group (MSC) and control group (CG). In the first, an intratesticular application of CTM was performed and in the CG only a buffer solution (PBS) was used. After the application, the animals were examined again. Fifteen days later, the animals were orchiectomized and histological analysis of the fragments were performed. Clinical examination showed no decline in the overall condition of the animals after application. There was no difference in the analysed variables, such as testicular volume, testicular surface temperature, ultrassonography, sperm kinetics, hormonal analysis and histology, in both groups after application. Thus, the results of the presente study support the hypothesis of the feasibility and safety of intratesticular application of allogeneic bone marrow MSC.For the second study, a group of 10 stallions submitted to scrotal thermal stress, testicular histological samples were analyzed before and after thermal insult, after intratesticular applications (PBS / MSC) and after 30 days of insult. The treated and control groups, after thermal stress, received mesenchymal stem cells and PBS respectively in their testicles. The method used to promote degeneration was efficient showing alterations in the seminiferous epithelium of both groups, however the treated group was able to recover a histological preview after 30 days of insult, while it was not observed in the control group. The characteristics that MSC has to stimulate tissue angiogenesis and proliferation, release of growth factors, eliminate free radicals, increase the capacity for renewal and cell differentiation, its immunomodulatory and immunosuppressive actions may be responsible for the outcome in the present study.It is concluded that the application of intratesticular mesenchymal stem cells is safe, in addition to promoting restoration of the initial patterns of the histological architecture of the testis in animals induced to testicular degeneration by thermal stress.
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Palavras-chave
garanhão, degeneração testicular, aplicação intratesticular, célula tronco mesenquimal, alogênica