Primary ovarian insufficiency refers to the loss of ovarian function before the age of 40 years and leads to amenorrhea and infertility. Primary ovarian insufficiency has diverse causes, but a common cause is exposure to gonadotoxic chemotherapy used in cancer treatment. Because of the risk for developing primary ovarian insufficiency, patients who want to preserve their fertility may consider various procedures for fertility preservation. However, current fertility preservation options are highly invasive, carry substantial risks, and have uncertain success rates. Recent studies from our group and others reported that mesenchymal stem cells and mesenchymal stem cell–derived exosomes can restore ovarian function in preclinical models of primary ovarian insufficiency by restoring damaged cells and inhibiting apoptosis. Although the restorative effect of mesenchymal stem cell–derived exosomes has been well reported in previous studies, the potential of mesenchymal stem cell–derived exosomes in preventing ovarian damage has not been fully elucidated. This study hypothesized that the antiapoptotic potential of mesenchymal stem cell–derived exosomes may protect ovarian tissue from chemotherapy-induced damage. In this study, we delivered mesenchymal stem cell–derived exosomes directly into the ovaries of mice before administration of chemotherapy. A total of 60 mice were divided into 3 groups (20 per group), which were labeled the control, chemotherapy, and fertility protection groups. Only the fertility protection group mice received exosomes, whereas the control and chemotherapy group mice received saline. After exosome injection, the chemotherapy and fertility protection groups of mice were subjected to chemotherapy to induce ovarian damage. After chemotherapy, we evaluated the protective effects of exosome treatment on ovarian function, such as estrous cyclicity, serum hormone levels, and the fertility rate, by comparing these outcomes between the chemotherapy and fertility protection groups. These outcomes were also compared with those of the control group for comparison with outcomes under healthy conditions. After intraovarian injection of exosomes before chemotherapy, the mice were able to maintain their estrous cycle (4- to 5-day cyclicity), serum anti-müllerian hormone level (66.06±26.40 ng/mL, not significantly different from that of the healthy controls), folliculogenesis (32.2±11.3 in the chemotherapy group vs 46.4±14.1 in the fertility protection group; <.05), expression of the steroidogenic acute regulatory protein gene (a the steroidogenesis marker) (0.44±0.11-fold expression in the chemotherapy group and 0.88±0.31-fold expression in the fertility protection group; <.05), and fertility (2 of 8 in the chemotherapy group and 5 of 8 in the fertility protection group), thereby showing prevention of chemotherapy-induced damage. We found that exosome treatment before chemotherapy can preserve ovarian function and protect fertility through the overexpression of ATP synthase-binding cassette transporters, such as ABCB1b (10.17±17.75-fold expression in the chemotherapy group and 44.14±33.25-fold expression in the fertility protection group; <.05) and ABCC10 (3.25±0.59-fold expression in the chemotherapy group and 5.36±1.86-fold expression in the fertility protection group; <.05). In this study, we present a novel fertility protection method using mesenchymal stem cell–derived exosomes. We concluded that mesenchymal stem cell–derived exosomes are a promising and simple treatment option for fertility protection in reproductive-aged patients who are receiving gonadotoxic chemotherapy.

中文翻译：

---

生育保护：一种使用间充质干细胞外泌体预处理来预防小鼠模型化疗引起的卵巢损伤的新方法

原发性卵巢功能不全是指40岁之前卵巢功能丧失，导致闭经和不孕。原发性卵巢功能不全有多种原因，但一个常见的原因是接触癌症治疗中使用的性腺毒性化疗。由于存在原发性卵巢功能不全的风险，想要保留生育能力的患者可以考虑采取各种保留生育能力的手术。然而，目前的生育力保存方案具有高度侵入性，存在巨大风险，并且成功率不确定。我们小组和其他人的最新研究报道，间充质干细胞和间充质干细胞衍生的外泌体可以通过恢复受损细胞和抑制细胞凋亡来恢复原发性卵巢功能不全临床前模型中的卵巢功能。尽管间充质干细胞衍生的外泌体的恢复作用在之前的研究中已得到充分报道，但间充质干细胞衍生的外泌体在预防卵巢损伤方面的潜力尚未完全阐明。这项研究假设间充质干细胞来源的外泌体的抗凋亡潜力可能会保护卵巢组织免受化疗引起的损伤。在这项研究中，我们在化疗前将间充质干细胞衍生的外泌体直接递送到小鼠的卵巢中。将60只小鼠分为3组（每组20只），分别标记为对照组、化疗组和生育保护组。只有生育保护组小鼠接受外泌体，而对照组和化疗组小鼠接受盐水。注射外泌体后，化疗组和生育保护组小鼠接受化疗诱导卵巢损伤。化疗后，我们通过比较化疗组和生育保护组之间的结果，评估了外泌体治疗对卵巢功能的保护作用，例如动情周期、血清激素水平和生育率。还将这些结果与对照组的结果进行比较，以与健康条件下的结果进行比较。化疗前卵巢内注射外泌体后，小鼠能够维持动情周期（4~5天周期），血清抗苗勒氏管激素水平（66.06±26.40 ng/mL，与健康对照无显着差异） ）、卵泡发生（化疗组为 32.2±11.3 vs 生育保护组为 46.4±14.1；<.05）、类固醇生成急性调节蛋白基因（类固醇生成标志物）的表达（化疗中表达量的 0.44±0.11 倍）组和生育保护组中的0.88±0.31倍表达；<.05)，以及生育力(化疗组中8个中的2个和生育保护组中8个中的5个)，从而显示出对化疗引起的损伤的预防。我们发现化疗前的外泌体治疗可以通过ATP合酶结合盒转运蛋白的过度表达来保留卵巢功能并保护生育力，例如ABCB1b（化疗组中表达10.17±17.75倍，生育保护组中表达44.14±33.25倍）组；<.05）和ABCC10（化疗组表达3.25±0.59倍，生育保护组表达5.36±1.86倍；<.05）。在这项研究中，我们提出了一种利用间充质干细胞衍生的外泌体的新型生育保护方法。我们的结论是，间充质干细胞衍生的外泌体是一种有前途且简单的治疗选择，可用于保护接受性腺毒性化疗的育龄患者的生育能力。