Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Department of Anatomy, School of Medicine, Shahrekord University of Medical Sciences, Shahrekord, Iran.
Ann Anat. 2023 Aug;249:152084. doi: 10.1016/j.aanat.2023.152084. Epub 2023 Mar 25.
Testicular torsion is considered a urological disorder that requires immediate detorsion surgery. Ischemia/reperfusion (I/R) injury after testicular torsion detorsion causes of drastic impairment of spermatogenesis and infertility. Cell-free-based approaches seem to be a promising strategy to prevent I/R injury, they have more stable biological properties, and they contain paracrine factors of mesenchymal stem cells. The purpose of this study was to evaluate the protective effects of human amniotic membrane derived mesenchymal stem cells (hAMSCs) secreted factors on mouse sperm chromatin condensation and spermatogenesis improvement after I/R injury. hAMSCs were isolated and characterized by RT- PCR and flow cytometry, preparation of hAMSCs secreted factors was performed. Forty male mice were randomly divided into 4 groups: sham-operated, torsion detorsion, torsion detorsion+ intratesticular injection of DMEM/F-12, and torsion detorsion+ intratesticular injection of hAMSCs secreted factors. After one cycle of spermatogenesis, the mean number of germ cells, Sertoli, Leydig, myoid as well as tubular parameters, Johnson score, and spermatogenesis indexes were evaluated by H& E and PAS stainings. Sperm chromatin condensation and relative expression of c-kit and prm 1 genes were assessed by aniline blue staining and real-time PCR, respectively. The mean number of spermatogenic cells, Leydig, myoid, Sertoli, spermatogenesis parameters, Johnson score, as well as germinal epithelial height and diameters of seminiferous tubules decreased significantly after I/R injury. The thickness of basement membrane and percentage of sperm with excessive histone significantly increased, while the relative expression of c-kit and prm 1 significantly decreased in torsion detorsion group (p 0.001). hAMSCs secreted factors remarkably restored normal sperm chromatin condensation, spermatogenesis parameters and histomorphometric organization of seminiferous tubules via intratesticular injection (p 0.001). Thus, hAMSCs secreted factors may potentially salvage torsion-detorsion-induced infertility.
睾丸扭转被认为是一种泌尿外科疾病，需要立即进行复位手术。睾丸扭转复位后发生的缺血/再灌注 (I/R) 损伤会导致精子发生严重受损和不育。无细胞治疗方法似乎是预防 I/R 损伤的一种有前途的策略，它们具有更稳定的生物学特性，并且含有间充质干细胞的旁分泌因子。本研究旨在评估人羊膜来源间充质干细胞 (hAMSCs) 分泌因子对 I/R 损伤后小鼠精子染色质浓缩和精子发生改善的保护作用。通过 RT-PCR 和流式细胞术分离和鉴定 hAMSCs，制备 hAMSCs 分泌因子。将 40 只雄性小鼠随机分为 4 组：假手术组、扭转复位组、扭转复位+睾丸内注射 DMEM/F-12 组和扭转复位+睾丸内注射 hAMSCs 分泌因子组。经过一个精子发生周期后，通过 H&E 和 PAS 染色评估生精细胞、支持细胞、间质细胞、肌样细胞以及管状参数、Johnson 评分和精子发生指数的平均值。通过苯胺蓝染色和实时 PCR 分别评估精子染色质浓缩和 c-kit 和 prm1 基因的相对表达。I/R 损伤后，生精细胞、间质细胞、肌样细胞、支持细胞、精子发生参数、Johnson 评分以及生精上皮高度和曲细精管直径均显著减少。基底膜厚度和具有过多组蛋白的精子百分比显著增加，而 c-kit 和 prm1 的相对表达显著降低（p<0.001）。通过睾丸内注射 hAMSCs 分泌因子可显著恢复正常的精子染色质浓缩、精子发生参数和曲细精管的组织形态学结构（p<0.001）。因此，hAMSCs 分泌因子可能潜在地挽救扭转复位引起的不育症。
