Department of Anatomical Sciences, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
Department of Tissue Engineering, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.
J Gynecol Obstet Hum Reprod. 2024 Nov;53(9):102828. doi: 10.1016/j.jogoh.2024.102828. Epub 2024 Jul 31.
Infertility is a significant public health concern affecting 10-15 % of couples. Young women undergoing gonadotoxic treatment are at higher risk of ovarian dysfunction and infertility. To mitigate this risk, ovarian tissue freezing and transplantation have been developed as a novel strategy. However, challenges such as follicular loss and dysfunction during the freezing process, and ovarian damage during transplantation, persist. This study aimed to investigate the potential of using appropriate antifreeze, antioxidant, wound healing, and biological hydrogels to reduce these injuries. Specifically, the effect of fibrin scaffold with endothelial cells and melatonin on apoptotic gene expression and antioxidants in cryopreserved ovaries after transplantation was examined.
A total of 36 adult female wistar rats) 6-8-week-old and weighing from 200 to 220 g) were divided equally into six groups (n = 6): 1) control group (C), 2) transplanted ovarian tissue after vitrification and thawing process (Group 1), 3) transplanted vitrified/thawed ovarian tissue while encapsulated in Fib/Alg hydrogel (Group 2), 4) transplanted vitrified/thawed ovarian tissue while encapsulated in Fib/Alg hydrogel in addition with melatonin (Group 3), 5) transplanted vitrified/thawed ovarian tissue while encapsulated in Fib/Alg hydrogel in addition with endothelial cells (Group 4) and 6) transplanted vitrified/thawed ovarian tissue while encapsulated in Fib/Alg hydrogel in addition with melatonin endothelial cells (Group 5). The ovaries were auto-transplanted in the rats' lumbar region. After 14 days, the ovaries were removed. Antioxidant levels (SOD, GPx, MDA, and TAC) were evaluated using ELISA, and apoptotic gene expressions (Bax/Bcl2 and caspase 3) were analyzed by real-time RT-PCR to determine apoptosis.
In the transplanted frozen ovary group, Bax/Bcl2 and caspase 3 gene expression increased significantly (P < 0.05), while antioxidant levels (SOD, GPx, MDA, and TAC) decreased. The encapsulated frozen ovary group showed decreased gene expression and increased antioxidant levels. The ovary group encapsulated with fibrin scaffold, endothelial cells, and melatonin had the most significant decrease in gene expression and increase in antioxidant levels (P < 0.05).
Coordinated action of Fibrin-based scaffold with endothelial cells and melatonin could decrease apoptosis gene expression and increase antioxidant levels in cryopreserved ovaries after transplantation, providing valuable insights into preserving fertility in young women undergoing gonadotoxic treatment.
不孕是一个严重的公共健康问题，影响了 10-15%的夫妇。接受性腺毒性治疗的年轻女性患卵巢功能障碍和不孕的风险更高。为了降低这种风险，已经开发出卵巢组织冷冻和移植作为一种新策略。然而，在冷冻过程中卵泡丢失和功能障碍以及移植过程中的卵巢损伤等挑战仍然存在。本研究旨在探讨使用适当的抗冻剂、抗氧化剂、伤口愈合和生物水凝胶来减少这些损伤的可能性。具体来说，研究了纤维蛋白支架与内皮细胞和褪黑素对移植后冷冻保存卵巢中凋亡基因表达和抗氧化剂的影响。
将 36 只成年雌性 wistar 大鼠（6-8 周龄，体重 200-220g）等分为六组（n=6）：1）对照组（C），2）玻璃化和解冻后移植卵巢组织组（第 1 组），3）玻璃化/解冻后移植卵巢组织并包裹在 Fib/Alg 水凝胶中组（第 2 组），4）玻璃化/解冻后移植卵巢组织并包裹在 Fib/Alg 水凝胶中添加褪黑素组（第 3 组），5）玻璃化/解冻后移植卵巢组织并包裹在 Fib/Alg 水凝胶中添加内皮细胞组（第 4 组）和 6）玻璃化/解冻后移植卵巢组织并包裹在 Fib/Alg 水凝胶中添加褪黑素和内皮细胞组（第 5 组）。将卵巢自体移植到大鼠的腰部。14 天后取出卵巢。使用 ELISA 评估抗氧化剂水平（SOD、GPx、MDA 和 TAC），通过实时 RT-PCR 分析凋亡基因表达（Bax/Bcl2 和 caspase 3）以确定凋亡。
在冷冻卵巢移植组中，Bax/Bcl2 和 caspase 3 基因表达显著增加（P<0.05），而抗氧化剂水平（SOD、GPx、MDA 和 TAC）下降。包裹冷冻卵巢组的基因表达下降，抗氧化剂水平升高。包裹纤维蛋白支架、内皮细胞和褪黑素的卵巢组的基因表达下降最显著，抗氧化剂水平升高最显著（P<0.05）。
纤维蛋白支架与内皮细胞和褪黑素的协同作用可降低移植后冷冻保存卵巢中的凋亡基因表达，增加抗氧化剂水平，为保护接受性腺毒性治疗的年轻女性的生育能力提供了有价值的见解。
