Muratori Monica, Tarozzi Nicoletta, Cambi Marta, Boni Luca, Iorio Anna Lisa, Passaro Claudia, Luppino Benedetta, Nadalini Marco, Marchiani Sara, Tamburrino Lara, Forti Gianni, Maggi Mario, Baldi Elisabetta, Borini Andrea
From the Department of Experimental, Clinical and Biomedical Sciences, Unit of Sexual Medicine and Andrology, Center of Excellence DeNothe, University of Florence (MM, MC, ALI, CP, BL, SM, LT, GF, MM, EB); Tecnobios Procreazione, Centre for Reproductive Health, Bologna (NT, MN, AB); and Clinical Trials Coordinating Center, AOU Careggi, Istituto Toscano Tumori, Florence (LB), Italy.
Medicine (Baltimore). 2016 May;95(20):e3624. doi: 10.1097/MD.0000000000003624.
Predicting the outcome of in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) is one main goal of the present research on assisted reproduction. To understand whether density gradient centrifugation (DGC), used to select sperm, can affect sperm DNA integrity and impact pregnancy rate (PR), we prospectively evaluated sperm DNA fragmentation (sDF) by TUNEL/PI, before and after DGC. sDF was studied in a cohort of 90 infertile couples the same day of IVF/ICSI treatment. After DGC, sDF increased in 41 samples (Group A, median sDF value: 29.25% [interquartile range, IQR: 16.01-41.63] in pre- and 60.40% [IQR: 32.92-93.53] in post-DGC) and decreased in 49 (Group B, median sDF value: 18.84% [IQR: 13.70-35.47] in pre- and 8.98% [IQR: 6.24-15.58] in post-DGC). PR was 17.1% and 34.4% in Group A and B, respectively (odds ratio [OR]: 2.58, 95% confidence interval [CI]: 0.95-7.04, P = 0.056). After adjustment for female factor, female and male age and female BMI, the estimated OR increased to 3.12 (95% CI: 1.05-9.27, P = 0.041). According to the subgroup analysis for presence/absence of female factor, heterogeneity in the association between the Group A and B and PR emerged (OR: 4.22, 95% CI: 1.16-15.30 and OR: 1.53, 95% CI: 0.23-10.40, respectively, for couples without, n = 59, and with, n = 31, female factor).This study provides the first evidence that the DGC procedure produces an increase in sDF in about half of the subjects undergoing IVF/ICSI, who then show a much lower probability of pregnancy, raising concerns about the safety of this selection procedure. Evaluation of sDF before and after DGC configures as a possible new prognostic parameter of pregnancy outcome in IVF/ICSI. Alternative sperm selection strategies are recommended for those subjects who undergo the damage after DGC.
预测体外受精（IVF）/卵胞浆内单精子注射（ICSI）的结果是目前辅助生殖研究的一个主要目标。为了解用于选择精子的密度梯度离心（DGC）是否会影响精子DNA完整性并影响妊娠率（PR），我们在DGC前后通过TUNEL/PI法对精子DNA片段化（sDF）进行了前瞻性评估。在90对不育夫妇进行IVF/ICSI治疗的同一天对sDF进行了研究。DGC后，41个样本中的sDF增加（A组，sDF中位数：DGC前为29.25%[四分位间距，IQR：16.01 - 41.63]，DGC后为60.40%[IQR：32.92 - 93.53]），49个样本中的sDF减少（B组，sDF中位数：DGC前为18.84%[IQR：13.70 - 35.47]，DGC后为8.98%[IQR：6.24 - 15.58]）。A组和B组的PR分别为17.1%和34.4%（优势比[OR]：2.58，95%置信区间[CI]：0.95 - 7.04，P = 0.056）。在对女性因素、男女年龄和女性BMI进行调整后，估计的OR增加到3.12（95%CI：1.05 - 9.27，P = 0.041）。根据有无女性因素的亚组分析，A组和B组与PR之间的关联出现了异质性（对于无女性因素的夫妇，n = 59，OR：4.22，95%CI：1.16 - 15.30；对于有女性因素的夫妇，n = 31，OR：1.53，95%CI：0.23 - 10.40）。本研究提供了首个证据，即DGC程序会使约一半接受IVF/ICSI的受试者的sDF增加，这些受试者随后的妊娠概率要低得多，这引发了对该选择程序安全性的担忧。评估DGC前后sDF可作为IVF/ICSI妊娠结局可能的新预后参数。对于那些在DGC后受到损害的受试者，建议采用替代的精子选择策略。
