Heidari Mahnaz, Darbandi Mahsa, Darbandi Sara, Sadeghi Mohammad Reza
Reproductive Biotechnology Research Center, Avicenna Research Institute (ARI), ACERCR, Tehran, Iran.
Turk J Urol. 2020 Sep;46(5):348-353. doi: 10.5152/tud.2020.19262. Epub 2020 May 27.
Sperm DNA fragmentation and maturation directly interferes with reproductive efficiency. Although there are several methods for assessing sperm DNA integrity, however, many of them are laborious and require high-precision equipment in the clinics. Thus, evaluating economic and reliable methods to prepare suitable sperm for assisted reproductive technologies without DNA damage is critical.
A total of 114 semen samples were collected and analyzed using computer-assisted semen analysis. The DNA fragmentation index (DFI) of all samples was evaluated by two methods of sperm chromatin dispersion (SCD) and sperm chromatin structure assay (SCSA). Besides, chromatin maturation index (CMI) was assessed by three methods including aniline blue (AB)-sperm chromatin maturation assay (SCMA), fluorescence microscopic chromomycin A3 (fmCMA3), and flow cytometric CMA3 (fcCMA3).
The result showed that the DFI had no statistically significant differences (p>0.05) between SCSA (26.98%±1.28%) and SCD (27.88%±1.278%), although SCD demonstrated a strong correlation with DNA maturity (p<0.001), which had not been seen in SCSA. Besides, the CMI demonstrated significant differences (p<0.001) when assessed by AB-SCMA (14.86%±0.65%), fmCMA3 (29.18%±1.01%), and fcCMA3 (22.45%±0.62%). Among these, only the fmCMA3 showed a significant correlation with semen parameters (p<0.01) and embryo development (p<0.001).
It seems that SCD and fmCMA3 were more accessible, affordable, and reliable tests for assessing DFI and CMI. It appeared these two methods may be the best choices for evaluating sperm DNA integrity in clinics.
精子DNA碎片化和成熟度直接影响生殖效率。尽管有多种评估精子DNA完整性的方法,但其中许多方法操作繁琐,且在临床中需要高精度设备。因此,评估经济可靠的方法来制备适用于辅助生殖技术且无DNA损伤的精子至关重要。
共收集114份精液样本,并使用计算机辅助精液分析进行分析。通过精子染色质扩散(SCD)和精子染色质结构分析(SCSA)两种方法评估所有样本的DNA碎片化指数(DFI)。此外,通过三种方法评估染色质成熟指数(CMI),包括苯胺蓝(AB)-精子染色质成熟分析(SCMA)、荧光显微镜下的放线菌素A3(fmCMA3)和流式细胞术CMA3(fcCMA3)。
结果显示,SCSA(26.98%±1.28%)和SCD(27.88%±1.278%)之间的DFI无统计学显著差异(p>0.05),尽管SCD与DNA成熟度呈强相关(p<0.001),而SCSA未观察到这种相关性。此外,通过AB-SCMA(14.86%±0.65%)、fmCMA3(29.18%±1.01%)和fcCMA3(22.45%±0.62%)评估时,CMI存在显著差异(p<0.001)。其中,只有fmCMA3与精液参数(p<0.01)和胚胎发育(p<0.001)呈显著相关。
似乎SCD和fmCMA3是评估DFI和CMI更易获得、更经济且更可靠的检测方法。看来这两种方法可能是临床评估精子DNA完整性的最佳选择。