Positive effect of resveratrol against preantral follicles degeneration after ovarian tissue vitrification
Introduction
Cryopreservation of ovarian tissue followed by transplantation procedure has been used as a strategy to preserve the fertility potential of human and animals [1,2]. However, the cell functionality of remaining structures in cryopreserved ovarian tissue is essential for a potential clinical benefit [[3], [4], [5]].
The vitrification technique has become routinely applied for gamete preservation in different species (murine [6]; ovine [7]; caprine [8]; bovine [9]; swine [10]; primate [11]; and human [12]). However, during the vitrification procedure reactive oxygen species (ROS) may be generated by different mechanisms, such as increased oxidative metabolism, osmotic stress, or even by changes in cell defense mechanisms [13,14]. The ROS levels seem to increase, especially after cell warming, once oxygen is reintroduced in cryopreserved cells and consequently cause oxidation and reduction reactions that lead to its production [15].
Studies have been carried out in order to minimize the cell damage induced by cryopreservation such as apoptotic activation, accumulate of reactive oxygen species (ROS), efflux of glutathione, and progressive DNA and cell membrane damage [16,17]. In this way the resveratrol, a polyphenol synthesized by many plants, have several biological activities that can be applied in therapeutic purposes such as anti-carcinogenic, antioxidant, anti-inflammatory, protection of the cardiovascular system, anti-obesity, and reproductive disturbances (aging-related infertility and polycystic ovary syndrome) [[18], [19], [20], [21], [22]]. As an antioxidant, resveratrol protects the mitochondrial function by activation of sirtuin-1 gene (SIRT1), found in granulosa cells, cumulus cells, oocytes, and blastocysts [16,23]. Moreover, experimental studies showed that the resveratrol could protect against the reduction of fertility caused by oxidative stress [19] and ageing related changes [24].
Several species have been used animal models for studies with ovarian tissue (mare [25]; rat [19]; sheep [26]; and monkey [27]). Thus, these animals represent a comparative experimental model for genetic improvement within its specie and provide, at the same time, important information that may be applied to human reproductive clinical practice [28,29]. Ovaries of bovine fetuses over 180 days of gestation have preantral follicles in all developmental stages, enabling the assessment of vitrification effects on ovarian tissue. Therefore, based on biological activities of resveratrol, the present study evaluate whether the addition of resveratrol to the vitrification/thawing media improved the cryotolerance of preantral follicles enclosed in bovine ovarian fragments. The following end points were evaluated: ovarian tissue viability, ROS production, follicle and oocyte diameters, and preantral follicle morphology and class distribution.
Section snippets
Ovaries
Ovaries (n = 18) of bovine fetuses (180 days of gestation) were collected in a local slaughterhouse. The age of the fetuses was determined by the length (cm) from the atlanto-occipital joint to the origin of the tail [30]. After collection, ovaries were washed in alcohol 70% for 10 s, washed twice in PBS, and then transported to the laboratory in isothermal containers at 4 °C [31].
Experimental design
In the laboratory, the ovarian cortex was cut in 3 × 3 × 1 mm fragments and randomly distributed in the following
Results
A total of 53,547 follicles distributed in 880 histological sections were evaluated. Overall, the mean percentage of normal follicles was different among treatments (P < 0.05), and it was greater (P < 0.05) in the VIT + RESV compared to the VIT group (Table 1). Moreover, the proportion of normal follicles of VIT + RESV group was greater (P < 0.05) in primordial, transitional, and primary follicles compared with VIT group. Within the group vitrified with resveratrol, primordial follicles showed
Discussion
To our knowledge, this is the first study that evaluated the effects of ovarian fragments vitrification with resveratrol on tissue viability, ROS production, follicle and oocyte diameters, preantral follicle morphology, and classification.
The present study has shown that the overall percentage of morphologically normal follicles was 1.7-fold higher in the group of fragments vitrified with resveratrol than the group vitrified without the use of antioxidant. Previous studies reported the benefits
Acknowledgment
Research was supported by the Foundation for Research Support of the State of Minas Gerais (FAPEMIG, grant #APQ-01905-14), Brazil.
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Review of ovarian tissue cryopreservation techniques for fertility preservation
2022, Journal of Gynecology Obstetrics and Human ReproductionOvarian tissue features assessed in bovine fetuses after vitrification and xenotransplantation procedures
2021, Reproductive BiologyCitation Excerpt :Five replicates were performed for each treatment (each ovarian pair was considered as replicate). The vitrification procedure was adapted from a previous study [17] and was carried out with two vitrification solutions (vs): VS1, consisted of minimum essential medium (MEM) supplemented with 10 mg/mL bovine serum albumin (BSA), 0.25 M sucrose, 10 % ethylene glycol (EG), and 10 % dimethyl sulfoxide (DMSO); VS2, MEM supplemented with 10 mg/mL BSA, 0.25 M sucrose, 20 % EG, and 20 % DMSO. Ovarian fragments were exposed to VS1 (4 min) and then to VS2 (1 min) at room temperature (RT ∼25 °C).
Aloe vera increases mRNA expression of antioxidant enzymes in cryopreserved bovine ovarian tissue and promotes follicular growth and survival after in vitro culture
2021, CryobiologyCitation Excerpt :Together, these effects control the cellular senescence, apoptosis and altered cell signaling, which compromises the viability of cryopreserved tissue [5]. In this sense, studies have been conducted to minimizing cell damage induced by cryopreservation [12]. Among the strategies, addition of natural antioxidant to vitrification solution has been evaluated [13].
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2020, CryobiologyCitation Excerpt :In addition, reactive oxygen species (ROS) is produced during cryopreservation of OTs, which can destroy follicles and cause oxidative damage [19,28]. Studies have shown that ROS is one of the key factors affecting the cryopreservation effect of vitrification, which can induce apoptosis or dysfunction of cells, and ultimately lead to the decline of oocyte survival rate and development ability [7,16,18]. Therefore, addition of exogenous antioxidants in vitrified solution can reduce the production of ROS and the oxidative stress response [3,19,20].