Gynecology-Andrology Unit, Medical School, Institute of Experimental and Clinical Research, Catholic University of Louvain, UCLouvain, 1200 Brussels, Belgium.
Advanced Drug Delivery and Biomaterials Unit, Louvain Drug Research Institute, Catholic University of Louvain, UCLouvain, 1200 Brussels, Belgium.
Int J Mol Sci. 2019 Nov 20;20(23):5833. doi: 10.3390/ijms20235833.
Fertility preservation for prepubertal boys relies exclusively on cryopreservation of immature testicular tissue (ITT) containing spermatogonia as the only cells with reproductive potential. Preclinical studies that used a nude mice model to evaluate the development of human transplanted ITT were characterized by important spermatogonial loss. We hypothesized that the encapsulation of testicular tissue in an alginate matrix supplemented with nanoparticles containing a necrosis inhibitor (NECINH-NPS) would improve tissue integrity and germ cells' survival in grafts. We performed orthotopic autotransplantation of 1 mm³ testicular tissue fragments recovered form mice (aged 4-5 weeks). Fragments were either non-encapsulated, encapsulated in an alginate matrix, or encapsulated in an alginate matrix containing NECINH-NPs. Grafts were recovered 5- and 21-days post-transplantation. We evaluated tissue integrity (hematoxylin-eosin staining), germ cells survival (immunohistochemistry for promyelocytic leukemia zinc-finger, VASA, and protein-boule-like), apoptosis (immunohistochemistry for active-caspase 3), and lipid peroxidation (immunohistochemistry for malondialdehyde). NECINH-NPs significantly improved testicular tissue integrity and germ cells' survival after 21 days. Oxidative stress was reduced after 5 days, regardless of nanoparticle incorporation. No effect on caspase-dependent apoptosis was observed. In conclusion, NECINH-NPs in an alginate matrix significantly improved tissue integrity and germ cells' survival in grafts with the perspective of higher reproductive outcomes.
青春期前男孩的生育力保存完全依赖于含有精原细胞的未成熟睾丸组织（ITT）的冷冻保存，因为精原细胞是具有生殖潜能的唯一细胞。使用裸鼠模型评估人类移植 ITT 发育的临床前研究的特点是重要的精原细胞丢失。我们假设将睾丸组织包裹在含有含有坏死抑制剂（NECINH-NPS）的纳米颗粒的藻酸盐基质中，将改善移植物中组织完整性和生殖细胞的存活。我们对从小鼠（4-5 周龄）中回收的 1mm³睾丸组织片段进行了原位自体移植。组织片段要么不包裹，要么包裹在藻酸盐基质中，要么包裹在含有 NECINH-NPS 的藻酸盐基质中。在移植后 5 天和 21 天回收移植物。我们评估了组织完整性（苏木精-伊红染色）、生殖细胞存活（早幼粒细胞白血病锌指、VASA 和蛋白球样免疫组化）、细胞凋亡（活性-caspase 3 免疫组化）和脂质过氧化（丙二醛免疫组化）。NECINH-NPs 可显著改善移植后 21 天的睾丸组织完整性和生殖细胞存活。无论是否掺入纳米颗粒，在 5 天内均可降低氧化应激。未观察到对 caspase 依赖性细胞凋亡的影响。总之，藻酸盐基质中的 NECINH-NPs 可显著提高移植物的组织完整性和生殖细胞存活，有望获得更高的生殖结果。
