Avaliação hormonal indireta e estudo da preservação folicular em tecido ovariano autólogo transplantado para região inguinal em ratos

Ceschin, Artemis Marti; Biondo-Simões, Maria de Lourdes Pessole; Thomaz, Beatriz Angélica Charlotte; Totsugui, Joel Takashi

doi:10.1590/s0102-86502004000100004

Cited by 6 publications

(5 citation statements)

References 8 publications

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“…Although both protocols showed similar results in the histological analysis for follicular counts, the vitrification protocol was significantly better to preserve the ovarian stem cell population. Ceschin et al (2004) using IHC analysis with Ki-67 concluded that although both conventional freezing and vitrification were feasible methods for ovarian tissue cryopreservation, vitrification was associated with the recovery of a greater number of potentially viable primordial follicles in rats, similarly to ours conclusions.…”

Section: Discussionsupporting

confidence: 74%

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Ovarian tissue vitrification is more efficient than slow freezing to preserve ovarian stem cells in CF-1 mice

Terraciano¹,

Garcez²,

Berger³

et al. 2020

JBRA Assisted Reproduction

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Objective:The aim of this study was to investigate the efficacy of protocols for mice ovary cryopreservation to compare the differences in Mouse Vasa Homologue expression (a germline cell marker) and ovarian viability after vitrification or slow freezing.Methods: Female CF1 mice aged 40-45 days were randomly divided into three groups: Control, vitrification or slow freezing. Their ovaries were surgically removed, rinsed in saline solution and cryopreserved. For vitrification, we used a commercial protocol and for slow freeze, we used 1.5 M ethylene glycol (EG) as cryoprotectant. After that, the ovaries were processed for histological an immunohistochemical analysis, and counting of primordial, primary, pre-antral and antral follicles.Results: No significant difference was found in the proportion of high-quality primordial, primary and pre-antral follicles after thawing/warming in the slow freezing and vitrification groups. The immunohistochemistry for MVH antibody demonstrated that the slow freeze group had a higher number of unmarked cells (p=0.012), indicating a harmful effect on the MVH expression in the ovarian tissue, where the cell structure is complex.Conclusion: Although both protocols indicated similar results in the histological analysis of follicular counts, the vitrification protocol was significantly better to preserve ovarian stem cells, an immature germ cell population. These cells are able to self-renew having regeneration potential, and may be effective for the treatment of ovarian failure and consequently infertility.

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Section: Discussionsupporting

confidence: 74%

“…The females were submitted to vaginal cytological analysis before euthanasia, as described by Ceschin et al (2004), in order to confirm the estrous cycle stage. Vaginal suspensions were collected in 0.5% NaCl (0.25mL), and the smears were evaluated according to Cooper et al (1998).…”

Section: Estrous Cycle Detectionmentioning

confidence: 99%

Ovarian tissue vitrification is more efficient than slow freezing to preserve ovarian stem cells in CF-1 mice

Terraciano¹,

Garcez²,

Berger³

et al. 2020

JBRA Assisted Reproduction

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“…There are several strategies in the transplantation of avascular heterotopic ovary developed with the objective of minimizing the loss of number of follicles, due to the of I/R injury, since about half of the primordial follicles are lost in initial ischemia, persisting until the beginning of neoangiogenesis 17 .…”

Section: Resultsmentioning

confidence: 99%

The Study of Estrous Cycle Phases Female Rats With the Use of N-Acetylcysteine in Ovarian Graft

Vieira

Gomes

Montero³

2021

J. Health Scie.

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Ovarian graft may be the target of the biochemical effects of oxidative stress caused at the time of transplantation. In order to evaluate the effect of N-acetylcysteine on the ovarian graft, regarding the estrous cycle preservation , 50 female and virgin EPM-1 Wistar rats, weighing up to 250g, originating from CEDEME of UNIFESP, were kept in adequate sanitary conditions. , receiving their own food and water. Daily vaginal smears were performed to identify the estrous phase for 8 days. The animals were randomly distributed into 05 groups: 1st Group (GTx), saline was administered subcutaneously, 2nd (NAC 150mgKg), 3rd (NAC 300mg / Kg), 4th (NAC 600mg / Kg) and 5th (NAC 1200mg / Kg) , that were administered NAC subcutaneously on the abdominal face, 60 minutes before left unilateral ovarian transplantation in retroperitoneum and contralateral oophorectomy for purposes of histomorphological analysis, with colpocytological evaluation. Euthanasia was performed by means of anesthetic lethal dose in half of the animals on the 4th postoperative day, with a single vaginal smear collection and euthanasia on the rest of the animals, between the 14th and 16th days, after the material was collected in order to define the estrus phase. It was evaluated in the graft that the animals exhibited in all groups return of estrous cycle in the later phase of the post-transplant, with better definition of regular cycle in the highest dosages of N-acetylcysteine. N-acetylcysteine induced the return of the estrous cycle in the rats’ ovarian graft , mainly in the highest dosage, proving its effectiveness in revascularization of the tissue after ischemia and reperfusion. Keywords: Acetylcysteine. Reperfusion. Histocompatibility Antigens. Menstrual Cycle. ResumoO enxerto ovariano pode ser alvo dos efeitos bioquímicos do stress oxidativo causado no momento do transplante. Com o objetivo de avaliar o efeito da N-acetilcisteína no enxerto ovariano, quanto à preservação do ciclo estral, foram utilizados 50 ratos EPM-1 Wistar, fêmeas e virgens, pesando até 250g, originários do CEDEME da UNIFESP, mantidos em adequadas condições sanitárias, recebendo ração própria e água. Realizados esfregaços vaginais diários para identificação da fase estral durante 08 dias. Os animais foram distribuídos aleatoriamente em 05 grupos: 1º Grupo (GTx), administrada solução salina via subcutânea, 2º (NAC 150mgKg), 3º (NAC 300mg/Kg), 4º (NAC 600mg/Kg) e 5º (NAC 1200mg/Kg), aos quais foi administrada NAC por via subcutânea em face abdominal, 60 minutos antes do transplante unilateral esquerdo do ovário em retroperitônio e à ooforectomia contra-lateral para fins de análise histomorfológica, com avaliação colpocitológica. A eutanásia foi realizada por meio da dose letal do anestésico em metade dos animais no 4º dia de pós-operatório, realizado única coleta de esfregaço vaginal e a eutanásia no restante dos animais, entre o 14 º e 16º dia, após a coleta do material para definição da fase estro. Foi avaliado no enxerto que os animais apresentaram em todos os grupos retorno de ciclo estral na fase mais tardia do pós-transplante, com melhor definição de ciclo regular nas dosagens mais elevadas de N-acetilcisteína. A N-acetilcisteína induziu o retorno do ciclo estral no enxerto ovariano de ratas, principalmente na maior dosagem comprovando sua eficácia na revascularização do tecido após isquemia e reperfusão. Palavras-chave: Acetilcisteína. Reperfusão. Transplante. Ciclo Ovariano.

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“…The present findings corroborate previous studies that show a greater number of immature follicles in the early phases and a greater number of corpora lutea in the last phases of the evaluation. 29,30,33,34 Yang et al 29 noted that 90% of the total follicles are immature, after 2 days of the transplant and that on the seventh day, this number was reduced and the number of mature follicles increased. They also noted an increase in corpora lutea on the second day.…”

Section: Discussionmentioning

confidence: 99%

Progressive Evaluation of Apoptosis, Proliferation, and Angiogenesis in Fresh Rat Ovarian Autografts Under Remote Ischemic Preconditioning

et al. 2016

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This study evaluated the remote ischemic preconditioning (R-IPC) early and late repercussion on fresh ovarian transplants, aiming to assess a probable protective effect in ovarian follicular pool. Sixty Wistar EPM-1 rats were used, divided in 2 study groups: ovarian transplantation (Tx) and Tx + R-IPC, submitted to ovary transplant with or without R-IPC, respectively. These groups were subdivided according to the date for euthanasia: 4th, 7th, 14th, 21st, and 30th days of the postoperatory period. Morphology, morphometry, neoangiogenesis (vascular endothelial growth factor [VEGF]), proliferative activity (Ki-67), and apoptosis (cleaved caspase-3) were evaluated. Remote ischemic preconditioning was performed in the common iliac artery. Fresh autologous ovarian tissue was implanted integrally in the retroperitoneum. All animals showed resumption of estrous phase after ovary transplantation. Remote ischemic preconditioning attenuated the lesions progressively from the 7th day, with greater number of the immature follicles (14 days, P < .05), but didn't affect mature follicles and corpora lutea (P > .05). Immunohistochemical analyzes, taken as a whole, show that R-IPC benefic effect is more evident in the later periods of evaluation, when a greater proliferative activity (14, 21, and 30 days, P < .05) and lesser cell apoptotic activity (21 and 30 days, P < .05). The VEGF expression was similar in all times (P > .05). Remote ischemic preconditioning could have a benefic effect in the progressive evaluation of freshly grafted ovarian, especially on the latest phases of the posttransplant period. The 14th day was a landmark in the recuperation of the graft. Further investigations are necessary to determine the role of R-IPC in this scenario and its effect in frozen-thawed ovarian tissue.

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Avaliação hormonal indireta e estudo da preservação folicular em tecido ovariano autólogo transplantado para região inguinal em ratos

Abstract: Avaliação hormonal indireta e estudo da preservação folicular em tecido ovariano autólogo transplantado para região inguinal em ratos

Cited by 6 publications

References 8 publications

Ovarian tissue vitrification is more efficient than slow freezing to preserve ovarian stem cells in CF-1 mice

Ovarian tissue vitrification is more efficient than slow freezing to preserve ovarian stem cells in CF-1 mice

The Study of Estrous Cycle Phases Female Rats With the Use of N-Acetylcysteine in Ovarian Graft

Progressive Evaluation of Apoptosis, Proliferation, and Angiogenesis in Fresh Rat Ovarian Autografts Under Remote Ischemic Preconditioning

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