Antioxidant defense capacity of ovarian tissue after vitrification in a metal closed system

Abstract

ObjectiveThe present study analyzed the quality of bovine ovarian tissue after vitrification in a metal closed chamber, in terms of putative changes in tissue viability (lactate dehydrogenase -LDH- release), anti-oxidant defenses, and redox parameters caused by cryopreservation.MethodsSmall and large fragmented bovine ovarian tissue specimens were vitrified in a metal chamber. After rewarming, tissue samples were fixed or cultured for 48 hours. Glutathione (GSH), protein sulfhydryl content, Total Radical Trapping Antioxidant Potential (TRAP), and lactate dehydrogenase were analyzed immediately after rewarming and after tissue culture.ResultsNo changes in antioxidant parameters or viability of rewarmed tissue samples were found immediately or 48h after vitrification. The method of vitrification in a metal closed chamber used in this study preserved the quality of bovine ovarian tissue. Furthermore, our data showed that the size of the tissue specimens did not affect post-vitrification biochemical viability parameters.ConclusionsWe believe that the vitrification methodology employed in the present study is safe and effective, and should be evaluated for use in humans.

Morphological study on the effects of sample size on ovarian tissue vitrification

“…There was, however, a drop on stromal integrity after the 48-hour culture of the large tissue fragments. This was an unexpected finding considering that previous studies measuring the rate of tissue necrosis by lactate dehydrogenase levels, did not detect any difference between tissue size and between vitrified and fresh samples after 48 hours in culture ( Massignam et al ., 2018 ). One explanation for the decrease in tissue integrity in large samples in the present study may be that the histology technique used to analyze the samples should be adjusted for larger tissue fragments exposed to a cryopreservation process.…”

“…Early experiments by Ferreira et al (2010), were performed using the slow freezing procedure and, to our knowledge, there is no similar research on the impacts of tissue sample size on the follicular and stroma integrity after vitrification/rewarming technique. However, it has been shown that the antioxidant defense capacity and tissue viability have similar values for vitrified ovarian tissue cut in small or large fragments, immediately after rewarming and after a 48h culture (Massignam et al, 2018).…”

Static magnetic field assisted thawing improves cryopreservation of mouse whole ovaries

“…Additionally, synthetic and plant‐derived antioxidant substances, such as melatonin, resveratrol, and aloe vera extract, have been tested in OT cryopreservation 29,30 . Additionally, vitrification of OT using a metal containment system and found no changes in antioxidant parameters or viability of the rewarmed tissue samples 31 …”

“…29,30 Additionally, vitrification of OT using a metal containment system and found no changes in antioxidant parameters or viability of the rewarmed tissue samples. 31 The effectiveness of static magnetic field (SMF) in biological systems has long been proved. 32,33 SMF can penetrate biological tissues and interact directly with mobile charges, such as proteins and ions, as well as magnetic compounds within these systems.…”

Comparative study of fertility parameters in vitrified human spermatozoa in the presence or absence of EmbryORP ® : A novel antioxidant

“…The oxidation reduction potential (ORP) is a direct measure of the balance between ROS and antioxidants, and if unbalanced, this can lead to oxidative stress (Agarwal et al., 2016). ROS play a major role in sperm physiology and have been reported to have both, beneficial and harmful effects depending on their concentration and the antioxidant scavenging capacity of the cell, which is diminished by the cryopreservation medium (Liu et al., 2018; Massignam et al., 2018; Taylor et al, 2009). We have previously observed that the addition of EmbryORP ® reduced the ORP of cultured vitrified human embryos.…”