Evaluation of apoptosis and stress oxidative in cryopreserved ovary after graft in fibrin scaffold with endothelial cells and melatonin

Abstract

Infertility is considered as a public health problem that affects about 10-15% of couples.In order to preserve fertility in young women with cancer who undergo chemotherapy or radiation therapy and face the risk of ovarian failure and infertility, various methods such as ovarian tissue freezing and transplantation are used. One of the problems that occurs during freezing is that the possibility of losing follicles increases. Also, during the transplantation, due to the delay in Vasculogenesis and occurrence of hypoxia and ischemia, the ovary is damaged. In this study, appropriate antifreeze, antioxidant, wound healing and biological hydrogels were used to reduce such injuries. The aim of this study is to investigate the induction effect of fibrin scaffold along with endothelial cells and melatonin on the expression of apoptotic genes (Bax) / Blc2 and caspase 3) and antioxidants (SOD), GPx, MDA and TAC) in cryopreserved ovary after transplantation. Materials and Methods: In this experimental study, 36 adult female rats (8 weeks) underwent ovariectomy. Then their ovaries were divided into six groups: control, Vitrification/transplantation (V/T), V/T with fibrin scaffold (V/T-S), V/T with fibrin scaffold and melatonin (V/T-S-M), V/T with fibrin scaffold and endothelial cells (V/T-S-C), V/T with fibrin scaffold, endothelial cells, and melatonin (V/T-S-M-C). Finally, the ovaries autotransplantation in the rats' lumbar region. After 14 days, the ovaries were removed. The level of antioxidants (SOD, GPx, MDA and TAC) was evaluated by the ELISA technique, and expressions of Bax / Blc2 and caspase 3 were related to the apoptosis by real-time RT-PCR Result: The results of this research showed that the expression of Bax/Blc2 and caspase 3 genes increased significantly (P<0.05) in the transplanted frozen ovary group and decreased in the encapsulated frozen ovary group. This decrease in the expression of Bax/Blc2 and caspase3 genes in the ovary group encapsulated with fibrin scaffold along with endothelial cells and melatonin was more than other groups and had a significant difference with the transplant frozen ovary group (P<0.05). In the groups with melatonin, the expression of Bax/Blc2 and caspase 3 genes was lower compared to the group with fibrin scaffold and fibrin scaffold with endothelial cells. There was no significant difference in the expression of Bax and Bcl2 between the group with fibrin scaffold and fibrin scaffold with endothelial cells. In fact, the presence of endothelial cells did not significantly reduce the expression of apoptosis genes. Also, the results of this research showed that the level of antioxidants SOD, GPx, MDA and TAC decreased significantly (P<0.05) in the transplanted frozen ovary group and increased in the encapsulated frozen ovary group. This increase in the antioxidant level of the ovary group encapsulated with fibrin scaffold along with endothelial cells and melatonin was higher than other groups and had a significant difference with the transplanted frozen ovary group (P<0.05).