非阻塞性無精症患者的精漿細胞外囊泡（SPEV）的大小和數量在不同取精結果的族群間比較

鄭裕生¹、陳幸儀¹、林于巧^２、林宗彥^２、林世杰^３、

國立成功大學醫學院附設醫院泌尿部¹，成大斗六分院²，國立成功大學醫學院基礎醫學研究所^３

The comparison of seminal plasma extracellular vesicles (SPEV) profile in NOA patients with different sperm retrieval outcome

Yu-Sheng Cheng¹, Sin-Yi Chen¹, Yu-Chiao Lin², Tsung-Yen Lin², Shih-Chieh Lin³

Department of Urology, National Cheng Kung University Hospital¹; Department of Surgery, Division of Urology, National Cheng Kung University Hospital Dou-Liou samples Branch, Yunlin, Taiwan²; The Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan^３

Purpose: 

Seminal plasma extracellular vesicles (SPEVs) are secreted cell-derived membrane structures that have been proven to carry proteins, nucleic acid, and lipids, which can modify the constitution and biological function of sperm and seminal plasma. This study aimed to characterize seminal plasma extracellular vesicles in NOA patients with different sperm retrieval outcomes.

Materials and Methods:

From January 2021 to February 2023, 106 infertile men diagnosed with non-obstructive azoospermia (NOA) in the NCKUH andrology clinic agreed to collect semen samples for this study. The Institutional Review Board approved this study. Seminal plasma is harvested and stored in liquid nitrogen for subsequent exosome extraction. First, SPEVs were isolated from seminal cell-free fluid by size exclusion chromatography column. Next, the concentration and size distribution of SPEVs were performed with nanoparticle tracking analysis (NTA) by the NANOSIGHT machine. The Mann-Whitney test was utilized for SPEVs differences between the successful sperm retrieval group versus failed sperm retrieval group. The Kruskal-Wallis test calculated comparison in different histopathologic groups, and Dunn’s multiple comparison tests accessed group-group comparison. A p-value less than 0.05 was set as statistical significance.

Result(s): 

Among 106 NOA semen samples, sixty NOA patients received subsequent mTESE procedures. The successful sperm retrieval group has 29 patients, and failed sperm retrieval group has 31 patients. Overall, SRR was 48.3%. The result showed no difference of SPEVs' average size(P=0.51) and concentration(P=0.98) between successful and failed sperm retrieval groups. Regarding testicular histopathology, 27 were hypospermatogenesis (HS), 13 were maturation arrest (MA), 17 were Sertoli-cell only (SCO), and 3 were fibrosis. The result showed no significant difference of SPEVs' average size between the four histopathology groups (P=0.48). However, the SPEV concentration of the fibrosis group was significantly lower than the other three groups (P=0.049). 

Conclusion(s):

The result of our cohort indicated that the SPEVs' size and concentration might not be used as a non-invasive biomarker to predict sperm retrieval outcomes in NOA patients. However, the discrepancy of SPEV concentration between different histopathology suggested the future research direction targeting bio-content of SPEV in NOA patients.   

Keywords: Seminal plasma extracellular vesicles (SPEV); exosome; semen; NOA; male infertility