Elevated Sperm DNA Fragmentation Research Articles

Clinically Meaningful Improvements in Sperm DNA Fragmentation Severity in Infertile Men Treated with Superoxide Dismutase Supplementation: A Single-Center Experience.

Antioxidants are commonly used for the treatment of idiopathic male infertility. Previous studies have shown that antioxidants are able to improve sperm quality, but little is known about their impact on sperm DNA fragmentation (SDF). Preliminary findings showed that superoxide-dismutase (SOD)-based antioxidant plus hydroxytyrosol and carnosol (FertiPlus® SOD) therapy was associated with SDF improvement in a small cohort of infertile men. Therefore, we aimed to assess rates of and predictors of semen parameters and SDF improvements in infertile men treated with FertiPlus® SOD therapy (SOD+) or with other antioxidants without SOD (SOD-) in the real-life setting. Data from 60 consecutive infertile men with baseline SDF ≥ 30% and treated with SOD+ or SOD- for at least three months were analyzed. Clinical parameters and serum hormones were collected. Sperm parameters and SDF were requested at baseline and after SOD+ or SOD- treatment. Clinically meaningful SDF change after treatment was defined as SDF improvement >20% compared to baseline. Propensity score matching was performed to adjust for baseline differences between groups. Descriptive statistics were used to compare clinical and hormonal characteristics between SOD+ and SOD- groups. Semen characteristics were compared before and after treatment. Logistic regression models investigated the association between clinical variables and SDF improvement. Groups were similar in terms of clinical, serum hormones and semen parameters at baseline after matching. Compared to baseline, sperm progressive motility (17 (10-36)% vs. 27 (12-41)%) and normal morphology (2 (1-6)% vs. 4 (2-6)%) significantly improved after SOD+ treatment (all p < 0.01), but not after SOD-. SDF values significantly improved after treatment in both groups, compared to the baseline evaluation (all p < 0.01). However, SDF values were lower after SOD+ than SOD- treatment (30 (22-36)% vs. 37 (31-42)%, p = 0.01). Similarly, a clinically meaningful improvement in SDF at follow-up was more frequently found after SOD+ than SOD- treatment (76.7% vs. 20.0%, p = 0.001). Multivariable logistic regression analysis showed that SOD+ treatment (OR 5.4, p < 0.001) was an independent predictor of clinically meaningful SDF improvement, after accounting for age and baseline FSH values. This cross-sectional study showed that, in a cohort of primary infertile men with SDF ≥ 30%, SOD-based treatment was significantly effective in improving SDF compared to antioxidants without SOD. Approximately 80% of men treated with SOD+ achieved clinically meaningful improvement in SDF after three months of treatment. Sperm progressive motility and normal morphology also improved after SOD+ therapy but not after SOD-. These results suggest that SOD+ treatment could be considered an effective option for the management of idiopathic infertile men with elevated SDF.

Effect of varicocele and varicocelectomy on sperm deoxyribonucleic acid fragmentation rates in infertile men with clinical varicocele.

Varicocele is the leading cause of male infertility. It can affect sperm quantity and quality through various non-mutually exclusive pathophysiological mechanisms, mainly oxidative stress. Excessive production of reactive oxygen species may overwhelm the sperm's defenses against oxidative stress and harm the sperm's DNA. Excessive sperm DNA breaks, so-called sperm DNA fragmentation, result from the oxidative stress cascade and are commonly found in the ejaculates of men with varicocele and fertility-related issues. Measuring sperm DNA fragmentation can provide valuable information on the extent of harm and might help select candidates for surgical treatment. Varicocelectomy is beneficial for alleviating oxidative stress-associated infertility and improving sperm DNA integrity. However, reproductive outcomes of infertile men with elevated sperm DNA fragmentation rates and surgically treated varicoceles remain poorly studied, and there is a need for well-designed trials to determine the impact of sperm DNA fragmentation reduction on natural and medically assisted reproduction.

Vitamin D levels and human sperm DNA fragmentation: a prospective, cohort study

BackgroundIntracytoplasmic sperm injection (ICSI) has revolutionized the treatment of couples with male factor infertility but results remain suboptimal and suggest the need for further investigation into the molecular biology of spermatozoa. Vitamin D has been implicated in spermatogenesis and sperm function. Hypovitaminosis D has been associated with abnormal testicular function, including elevated sperm DNA fragmentation in a murine model. This study’s objective was to evaluate if there is a correlation between Vitamin D sufficiency and human spermatozoa DNA fragmentation index % (DFI%) in infertile couples.ResultsA prospective cohort study using a consecutive, convenience sample of subjects with infertility. The primary endpoint was the effect of Vitamin D sufficiency on human spermatozoa DFI%, and secondary outcomes included Vitamin D’s effect on moderate DFI%, high DFI%, High DNA stainability % (HDS%), sperm density (million/mL), sperm total motility (% total) and sperm strict morphology (% total). Of the 111 participating, 9 were excluded, leaving 102subjects. The subjects were stratified by vitamin D levels: deficient (< 20 ng/mL; n = 24), insufficient (20–30 ng/mL; n = 43),, and sufficient (> 30 ng/mL; n = 35). There were no statistical difference between the categories of serum vitamin D levels and sperm DFI% as well as the secondary outcomes. An increased BMI was associated with low serum vitamin D levels (p = 0.0012).ConclusionVitamin D deficiency was not associated with sperm DFI% or routine sperm parameters. Previous animal and human studies have demonstrated conflicting results between sperm parameters and Vitamin D levels. Redundant pathways in Vitamin D and calcium homeostasis in the human male reproductive tract may maintain essential reproductive processes during Vitamin D insufficiency or deficiency.Trial registrationTrial Registration Number: MOD00002311 (ClinicalTrials.gov).

Influence of sperm DNA fragmentation on the clinical outcome of in vitro fertilization-embryo transfer (IVF-ET).

PurposeTo evaluate the effect of elevated sperm DNA fragmentation index (DFI) on fresh and frozen embryo transfer cycles.MethodsA retrospective study was performed with 549 fresh embryo transfer cycles and 1340 frozen embryo transfer cycles after in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) from 2016 to 2021.ResultsThe statistical results of 549 fresh embryo transfer cycles showed that the delivery rate in the normal sperm DFI group (43.9% vs. 27.1%, P = 0.014) was significantly higher than that in the abnormal sperm DFI group, and there were no significant differences in the biochemical pregnancy rate (59.0% vs. 50.8%, P = 0.232), clinical pregnancy rate (53.1% vs. 40.7%, P = 0.072), or miscarriage rate (17.3% vs. 33.3%, P = 0.098) between the two groups. The results of 1340 frozen embryo transfer cycles showed that the biochemical pregnancy rate (57.9% vs. 45.6%, P = 0.006) and clinical pregnancy rate (50.3% vs. 40.7%, P = 0.027) in the normal sperm DFI group were significantly higher than those in the abnormal sperm DFI group. The delivery rate (40.9% vs. 33.3%, P = 0.074) and miscarriage rate (18.6% vs. 18.0%, P = 0.919) were not significantly different between the two groups.ConclusionThe increase of sperm DFI significantly reduced the delivery rate of fresh embryo transfer cycles and the biochemical pregnancy rate and clinical pregnancy rate of frozen embryo transfer cycles.

P-064 Clinical outcomes of 127 patients with recurrent implantation failure treated with testicular sperm aspiration (TESA)

Abstract Study question Are the embryological, clinical and newborn outcomes using aspirated testicular sperm improved in cases with recurrent implantation failure previously treated with ejaculated sperm? Summary answer Aspirated testicular sperm enabled to obtain significant higher embryological, clinical and newborn outcomes in cases with recurrent implantation failure previously treated with ejaculated sperm. What is known already High levels of sperm DNA fragmentation (SDF) were associated to poor clinical outcomes (1-Simon et al., 2017). Testicular sperm display lower SDF than ejaculated sperm (2-Sakas and Alvarez, 2010), improving clinical outcomes in cases with abnormal semen parameters (3-Awaga et al., 2018; 4-Kang et al., 2018), recurrent implantation failure (RIF) and pregnancy loss (RPL) (5-Esteves et al., 2017), and elevated SDF (6-Ambar et al., 2021). As only a few studies are specifically dedicated to RIF, we expanded the number of cases and first provided full demographic, stimulation, embryological, clinical and newborn outcomes. References 1-(https://doi.org/10.4103/1008-682X.182822); 2-(https://doi.org/10.1016/j.fertnstert.2009.10.046); 3-(https://doi.org/10.1016/j.rbmo.2018.08.017); 4-(https://doi.org/10.1038/s41598-018-26280-0); 5-(https://doi.org/10.1016/j.fertnstert.2017.06.018); 6-(https://doi.org/10.5534/wjmh.200084 Study design, size, duration We retrospectively evaluated during consecutive years (2010-2020) 63 patients with recurrent implantation failure, which accepted to perform testicular sperm aspiration (TESA) as an alternative treatment. These patients presented a long history of failed treatments (153 cycles) using ejaculated sperm. From these cycles, no pregnancy ensued. The present study compares 127 treatment cycles, 80 with testicular sperm (17 cases repeated TESA) and 47 with ejaculated sperm from the same patients performed at the present IVF clinic. Participants/materials, setting, methods Patients were screened for karyotype abnormalities, for Y-chromosome microdeletions (7-Gonçalves et al., 2016), and for SDF with the TUNEL assay (8-Sá et al., 2015). Conventional semen analysis was performed according to World Health Organization guidelines (9-WHO, 2010). Male evaluation and TESA was performed by the same experienced urologist (LF) according to established protocols (10-Madureira et al 2014). The procedure was performed entirely on an outpatient basis, with no complications reported. References 7-(https://doi.org/10.4103/1008-682X.172827); 8-(https://doi.org/10.1016/j.rbmo.2015.06.019); 9-(https://apps.who.int/iris/handle/10665/44261); 10-(https://doi.org/10.1111/j.2047-2927.2014.00231.x). Main results and the role of chance The mean ages were 35.5±3.4 (26-42)-female and 38.1±5.7 (29-59)-male. There were 4 abnormal karyotypes (3-female, 1-male), all without known relevance. Most cases had asthenozoospermia and teratozoospermia (65.1%), or oligoasthenoteratozoospermia (41.8%). Of the 19 cases with &amp;lt;5M/ml, none presented Y-chromosome microdeletions. Although we do not routinely perform SDF testing, 15 patients had previous SDF values (12, &amp;gt;20%; 8, &amp;gt;36%). Female basal characteristics and testicular evaluation were under normal values. The TESA procedure took about 15-20 min, and the time of laboratorial search around 30-60 min. Cases using testicular sperm showed significant higher rates of fertilization (64% vs 73%-p=0.005), blastocyst development (47% vs 62%-p=0.010), implantation (6% vs 27%-p=0.000), clinical pregnancy (10% vs 39%-p=0.001), live birth delivery (5% vs 28%-p=0.005) and newborn (5% vs 32%-p=0.000) than ejaculated sperm. No significant differences were observed regarding the rates of embryo cleavage (95% vs 94.8%) and high quality embryos (89.4% vs 94%), in the mean number of transferred embryos (1.8±0.4 vs 1.9±0.4), or in the abortion rate (2 cases-50% vs 7 cases-25.9%). Cases using testicular sperm had 22 frozen-thawed embryo transfer cycles, enabling per initiated cycle a cumulative pregnancy rate of 45%, live birth delivery rate of 31.3% and newborn rate of 37.5% (32 newborn). Limitations, reasons for caution Although presenting the higher number of cycles using TESA in the treatment of RIF, this number needs to be increased for drawing more definitive conclusions, as these women present a diversity of conditions, rendering subgrouping difficult. In the future, it would also be important to evaluate SDF in all cases. Wider implications of the findings In conclusion, the present results gave further evidence for the superiority of using testicular sperm instead of ejaculated sperm in cases with recurrent implantation failure. Data also evidences the security of using testicular sperm aspiration, as there were no pregnancy or delivery complications, or congenital anomalies among the 32 newborn. Trial registration number Not Applicable

Microfluidic preparation of spermatozoa for ICSI produces similar embryo quality to density-gradient centrifugation: a pragmatic, randomized controlled trial.

Does processing of spermatozoa for IVF with ICSI by a microfluidic sperm separation device improve embryo quality compared with density-gradient centrifugation? Patients randomized to microfluidic sperm preparation had similar cleavage- and blastocyst-stage embryo quality and clinical and ongoing pregnancy rates to those who underwent standard sperm processing for IVF with ICSI. Microfluidic sperm preparation can isolate spermatozoa for clinical use with minimal DNA fragmentation but with unclear impact on clinical outcomes. A prospective randomized controlled trial of 386 patients planning IVF from June 2017 through September 2021 was carried out. One hundred and ninety-two patients were allocated to sperm processing with a microfluidic sperm separation device for ICSI, while 194 patients were allocated to clinical standard density-gradient centrifugation (control) at an academic medical centre. In an intention to treat analysis, there were no differences in high-quality cleavage-stage embryo fraction [66.0 (25.8)% control versus 68.0 (30.3) microfluidic sperm preparation, P = 0.541, absolute difference -2.0, 95% CI (-8.5, 4.5)], or high-quality blastocyst fraction [37.4 (25.4) control versus 37.4 (26.2) microfluidic sperm preparation, P = 0.985, absolute difference -0.6 95% CI (-6, 5.9)] between groups. There were no differences in the clinical pregnancy or ongoing pregnancy rates between groups. The population studied was inclusive and did not attempt to isolate male factor infertility cases or patients with a history of elevated sperm DNA fragmentation. Microfluidic sperm separation performs similarly to density-gradient centrifugation in sperm preparation for IVF in an unselected population. No external funding to declare. M.P.R. is a member of the Clinical Advisory Board for ZyMōt® Fertility, Inc. NCT03085433. 21 March 2017. 16 June 2017.

O-210 Sperm DNA fragmentation (SDF) is not associated with adverse maternal and neonatal outcomes in IVF-ICSI cycles with autologous oocytes

Abstract Study question Does an elevated SDF (&amp;gt;15%) increase the odds of adverse maternal and neonatal outcomes in autologous oocyte IVF-ICSI cycles from unselected couples? Summary answer No adverse effects of high SDF on obstetric and neonatal outcomes have been found in couples with sperm fragmentation undergoing IVF-ICSI cycles with own eggs. What is known already Sperm chromatin integrity assessment has been implemented as an additional tool in the clinical evaluation of sperm quality in infertile patients undergoing an assisted reproduction treatment. All of the published reports to date appraise its effect on clinical outcomes, and how it impacts embryo quality and the pregnancy chances after IVF and ICSI cycles. Sperm DNA integrity has also been hypothesized to affect offspring health but not many studies have reported in humans if an elevated SDF raises the risks of obstetric, delivery and neonatal outcomes. Study design, size, duration Multicentric retrospective cohort study of all IVF-ICSI cycles using autologous oocytes between January 2000-March 2019 at Spain IVIRMA clinics of couples with a SDF test on their ejaculated semen. The sperm fragmentation index was measured in all men with TUNEL assay. The database included 228 couples which had a delivery with at least a newborn. Subjects were divided into two study groups according to their level of SDF: ≤15% (low SDF) or &amp;gt; 15% (high SDF). Participants/materials, setting, methods Patients with missed information on maternal and neonatal outcomes were not counted for the analysis. The obstetric outcomes were gestational age, gestational diabetes, preeclampsia (hypertension with proteinuria after 20 weeks of gestation) and type of delivery. Neonatal outcomes were sex, birth weight, length, head circumference, Apgar score at 1, 5, 10 minutes, and neonatal intensive care unit (NICU) admission. Student’s t-test and Fisher’s test were used for statistical analysis. A p-value&amp;lt;0.05 was considered statistically significant. Main results and the role of chance Maternal age mean was 37.4 years (95%CI 36.9-38.0) in ≤ 15%SDF group and 37.2 years (95%CI 36.1-38.4) in &amp;gt; 15%SDF group (p = 0.8). Similar gestational age was found, 41.8 weeks (95%CI 41.3-42.2) in ≤ 15%SDF and 41.3 weeks (95%CI 40.4-42.3) in &amp;gt; 15%SDF. Gestational diabetes incidence was higher in &amp;gt; 15%SDF compared to ≤ 15%SDF group (3.5% versus 1.7% (OR = 2.0 (95%CI 0.03-39.8), p = 0.5). Equally, the incidence of preeclampsia was 3.6% in patients with high SDF versus 1.7% in couples with low SDF, OR = 2.1 (95%CI 0.03-41.3), p = 0.5. Type of delivery frequency was in the ≤15%SDF group 61.9% vaginal and 38.1% cesarean, while in the &amp;gt;15%SDF group 62.1% vaginal and 37.9% cesarean (OR = 1.0 (95%CI 0.4-2.6), p = 1.0). The overall proportion of singleton pregnancies was 87.2% (95%CI 82.4-91.2) and twins 12.8% (95%CI 8.8-17.6). There were no statistically differences between groups in the rate of delivery of twins and in the sex ratio of the newborns. When comparing the newborns of ≤ 15%SDF with &amp;gt;15%SDF group, the average of weight was 3011.7g (95%CI 2912.2-3111.2) versus 2986.4g (95%CI 2753.1-3219.7), of length was 49.2cm (95%CI 48.3-50.0) versus 49.5cm (95%CI 49.2-49.9), of head circumference was 34.9cm (95%CI 34.6-35.2) versus 34.3cm (95%CI 33.4-35.2). No statistically differences were observed for Apgar punctuation and for NICU admission. Limitations, reasons for caution Due to the retrospective nature of the study we have missing data from the lack of follow-up of many patients after the confirmation of the ongoing pregnancy. Although pregnancies of couples with elevated SDF have a higher incidence of gestational diabetes and preeclampsia, the sample size evaluated is a limitation. Wider implications of the findings This is one of the first reports to evaluate the relationship between paternal DNA damage and obstetric risks and neonatal health in couples with high SDF who underwent IVF-ICSI in our centers. Despite SDF did not jeopardize the maternal and neonatal outcomes, more studies are needed to confirm this conclusion. Trial registration number NA

P–030 A sperm chromatin damage &amp;gt;15% negatively impacts on the quality of embryos obtained from ovum donation ICSI cycles of unselected couples

Abstract Study question Is embryo quality downgraded in couples with elevated sperm DNA fragmentation (SDF) in the ejaculated semen of male partner using donated eggs? Summary answer The rate of good quality embryos at day 3 and blastocyst-stage is statistically inferior in males with SDF&amp;gt;15% undergoing ICSI cycles with donated oocytes. What is known already The effect of a damaged paternal chromatin will be shown from the 8-cell stage of embryo development, a time which the genome of the embryo is transcriptionally active. Fertilization with a spermatozoon with fragmented DNA may impair the quality of the embryos obtained per cycle, and therefore reduce the chances of pregnancy. The use of donated oocytes is an ideal model to evaluate the real effect of SDF on embryo quality by standardizing the female factor. In addition, we have a large cohort of ovum donation cases in our history, which allows a more proper evaluation of the effect. Study design, size, duration Retrospective multicentric study including the clinical data of 864 couples of ovum donation program who underwent 1903 ICSI cycles between January 2000 and March 2019. The DNA fragmentation of their ejaculated spermatozoa was measured by TUNEL assay (Terminal deoxynucleotidyl transferase dUTP nick end labeling). Two study groups were created according to the SDF level: ≤15% (low) (n = 1626) or &amp;gt; 15% (high) (n = 277). Participants/materials, setting, methods Embryos were evaluated throughout embryonic development according to classical morphological parameters at day 3 (D3), on cleavage-stage, and at day 5 (D5), on blastocyst-stage (trophectoderm (TE) and inner cell mass (ICM)), following ASEBIR guidelines, categorized from A to D. Embryos scored as A and B were considered to be good quality. The proportion of embryos was calculated according to the total number of correctly fertilized oocytes or zygotes. A p &amp;lt; 0.05 was considered significant. Main results and the role of chance A total of 6130 embryos were evaluated. The SDF average of ≤ 15% group was 5.9% (95%CI 5.7–6.1) and 24.3% (95%CI 23.2–25.3) in the &amp;gt;15% group. The cycle-related characteristics and the seminal parameters were comparable. The proportion of optimal cleavage-stage embryo (number of A+B embryos at D3) per cycle was 21.7% (95%CI 19.0–24.5) (8.1 average cells number, 0.8 embryo fragmentation average, symmetry 1, mononucleated cells) in ≤ 15% SDF group versus 21.1% (95%CI 13.9–28.3) (8.2 cells number average, 1.3 embryo fragmentation average, symmetry 1, mononucleated cells) (p &amp;lt; 0.001). The blastocyst-stage arrival rate (number of embryos at D5) per cycle was higher in the &amp;gt;15% SDF group (p &amp;lt; 0.001), 53.4% (95%CI 48.8–58.1) (TE quality A:20.5%, B:42.5%, C:22.7%, D:14.8%, and the ICM quality A:26.1%, B:52.1%, C:13.2%, D:6.2%) versus 49.9% (95%CI 48.1–51.6) (TE quality A:21.1%, B:42.8%, C:21.85, D:14.1% and ICM A:26.6%, B:55.5%, C:11.1%, D:4.7%) in the low SDF group. The rate of good quality blastocyst (number of quality A+B embryos in D5) per cycle was significantly higher in the couples with low SDF (24.8% (95%CI 23.6–25.9)) than in those with elevated SDF (23.5% (95%CI20.9–26.2)) (p &amp;lt; 0.001). Accordingly, the A+B blastocyst rate divided by the total number of blastocysts was 59.1% (95%CI 56.7–61.4) versus 55.9% (95%CI 49.9–62.0) (p &amp;lt; 0.001), respectively. Limitations, reasons for caution The main limitation is that retrospective design of the study may not eliminate the potential unaccounted-for bias derived from the clinical practice of multiple centers even though both groups were statistically comparable. Also, the assessment of embryo quality is still remaining highly subjective to embryologists. Wider implications of the findings: Although the effect size is small, it may be useful in clinical practice when an ICSI cycle yields no good-quality embryos, as one of the underlying causes of that fact. Knowing the SDF level can be a helpful tool in making subsequent clinical decisions aimed at improving outcomes for couples. Trial registration number Not applicable

P–065 Advanced paternal age does affect egg donation program outcomes?

Abstract Study question Does increasing paternal age has a negative impact in fertilization (FR), blastulation (BR), clinical pregnancy (CPR) and miscarriage (MR) rates in an egg donation program? Summary answer The increase paternal age in an egg donation program has not a negative impact in fertilization rate, blastulation rate, clinical pregnancy rates and miscarriage rates. What is known already It is well documented that semen quality is affected with increasing paternal age but there is no evidence-based definition of what is advanced paternal age. There is controversial information about if the increasing paternal age affects in vitro fertilization results, and when this negative impact could begin. Study design, size, duration This was a single center retrospective cohort study, involving 485 first single embryo transfer of an egg donation program, from January 2017 to December 2019. Participants/materials, setting, methods All first embryo transfer of egg donation cycles performed at CEGyR, Buenos Aires, Argentina were included. Elevated sperm DNA fragmentation (TUNEL &amp;gt;20), sperm bank, and testicular biopsy cycles were excluded. Patients were divided according to male partner age: (1) &amp;lt;41, (2) 41–44, (3) 45–50 and (4) &amp;gt;50 years old. Group (1) was considered the control group. Statistical analyses were performed for FR and BR with ANOVA and CPR and MR with chi-squared tests. Main results and the role of chance The number of patients in group (1) was 200, in (2) 130, in (3) 117 and in (4) 38. Male average age was 36,8 in group (1), 42,2 in (2) 47,1 in (3) and 54,2 in group (4). The FR in group (1) was 72,60%, in group (2) was 73%, in (3) was 75% and 73% in (4). ANOVA results for FR: F = 0,65 (p: 0,58). The BR, defined as the relation between the total number of blastocysts over the number of fertilized oocytes in a cycle, was in group (1) 46,35%, in group (2) was 45%, in group (3) 46%, and in group (4) 42%. ANOVA results for BR F = 0,36 (p:0,78). The CPR in group (1) was 42,19%. Comparing with the other groups: group (2) was 37,09% (chi-square statistic=0,64 p:0,43); group (3) 34,58% (2,32 p:0,13); and group (4) was 32,43% (1,48 p:0,22). The MR in group (1) was 12,49%. Comparing with the other groups: group (2) was 18,55% (chi-square statistic=2,31 p:0,12); group (3) 14,94% (1,01 p:0,32); and group (4) was 15,85% (0,91 p:0,33). For all results analyzed there were not a statistically difference between groups. Limitations, reasons for caution The main limitation of this study was its retrospective design based on data from a single center which may be subject of bias. Wider implications of the findings: Further large prospective studies are required to make meaningful comparisons. Our findings give no support for a general recommendation. Trial registration number Not applicable

Elevated sperm DNA fragmentation does not predict recurrent implantation failure.

In this study, we sought to determine whether sperm DNA fragmentation (DFI%) and high DNA stainability (HDS%) evaluated by sperm chromatin structure assay (SCSA) predict recurrent implantation failure (RIF) or pregnancy rate. A retrospective study was performed of consecutive cycles of ICSI treatment from 2009 to 2018. A total of 386 couples that underwent 1,216 frozen embryo transfer (FET) cycles were analysed. Mean female and male age was 34±3.6years and 37.3±6.6years, respectively, and a median total motile sperm count (TMSC) was 43.5 [9.9-105.5] million. Overall median DFI% and HDS% was 12 [7.1-18.9] and 9.6 [6.5-14.4] respectively. On multivariable analysis, DFI% and HDS% were not associated with RIF (DFI%: OR=1.01, 95% CI: 0.98-1.04, p=.414; HDS%: OR=0.97, 95% CI: 0.94-1.01, p=.107) or IVF success, defined as clinical pregnancy (DFI%: OR=1.00, 95% CI: 0.99-1.01, p=.641; HDS%: OR=1.01, 95% CI: 0.99-1.02, p=.565). We found that neither DFI% or HDS%, as assessed by SCSA, were predictive of RIF or pregnancy rate. This finding suggests that sperm DNA fragmentation does not predict RIF or pregnancy rate.

Efficacy of varicocelectomy for sperm DNA integrity improvement: A meta-analysis.

Varicocele is common among male adolescents and adults. Varicocelectomy is the major means of varicocele repair. There is evidence that varicocelectomy could decrease sperm DNA fragmentation. However, studies evaluating the efficacy of varicocelectomy for sperm DNA integrity usually have a small sample size, and there is no up-to-date meta-analysis in this area. The present meta-analysis is to evaluate the efficacy of varicocelectomy for sperm DNA integrity. A literature search was conducted to identify all relevant studies from Medline database (PubMed), Cochrane Library, and OVID Embase from the inception dates to 08 June 2020. A total of 11 prospective studies including 394 cases were included in this meta-analysis. All analysis was performed using Stata version 16.0. In the random-effect model for 11 studies, DNA fragmentation index (%) of clinical varicocele patients decreased by 5.79 (95% CI, -7.39 to -4.19) after varicocelectomy. While after excluding one study with high heterogeneity, from the results of fixed-effect model, DNA fragmentation index decreased by 6.14 (95% CI, -6.90 to -5.37) on average. Sperm DNA integrity of clinical varicocele patients was significantly improved following varicocelectomy. Therefore, it is necessary to include elevated sperm DNA fragmentation index as a molecular indicator for varicocelectomy among clinical varicocele cases.

Covid-19: A possible emerging impact on male fertility

SARS-CoV-2 is an emerging pandemic; main complementary protein and mediator for SARS-CoV-2 has been identified to be the angiotensin converting enzyme 2 (ACE2) that exists in various organs and highly expressed within the testicular tissue (spermatogonia, Leydig and Sertoli cells). SARS-CoV-2 binds to ACE2 and TMPRSS2 forming protein complexes mediating viral replication within the male gonads. Hence, an impact on testicular tissue could be observed post-infection. The structure of the sperm glycocalyx could serve as an adequate carrier for SARS-CoV-2. SARS-CoV-2 complex protein formation triggers an elevated immune response in a “cytokine storm” like action. Such leads to extensive tissue damage mediated by TFN-α, IL-1β, IL-18 and IFN-γ. ROS elevation is noted during the infection from the viral-induced cellular damage. Such aggressive immune response, ROS accumulation, complex proteins formation and renin system failure could also progress to intercellular oedema which could lead to intratesticular obstruction with forming consequences including sperm count reduction and elevated Sperm DNA Fragmentation Index (SDFI). Testicular ultrasonography, semen analysis and male hormone profiling are recommended. Post recovery repetition for the previously mentioned tests would provide a useful insight into the andrological observations. Semen cryopreservation is highly recommended for vulnerable patients. Semen purification techniques could be adopted prior to cryopreservation and handling.

SC24 - Clomiphene citrate and FSH treatment in men with elevated sperm DNA fragmentation index: Findings from a cross-sectional study

SC24 - Clomiphene citrate and FSH treatment in men with elevated sperm DNA fragmentation index: Findings from a cross-sectional study

Free circulating DNA and DNase activity in the ejaculates of men with spinal cord injury.

Retrospective descriptive study. To study the presence of cell-free DNA (cfDNA) and DNase activity in males with spinal cord injury (SCI) with elevated sperm DNA fragmentation. Hospital in Toledo, Spain; University-based Genetics laboratory in Madrid, Spain. Semen was collected from 15 males with spinal cord injury and elevated sperm DNA fragmentation (SDF). The presence and concentration of cfDNA was assessed using standard gel electrophoresis and microfluidic electrophoresis. DNase activity was evaluated in seminal plasma and under the presence of EDTA and EGTA to control the response of enzyme activity. cfDNA fragments were mapped on the sperm genome using FISH. All results were compared to 15 normozoospermic fertile donors. Standard gel electrophoresis revealed a cfDNA band of ~150 bp in all samples from males with SCI; this band was ocasionally accompanied by another band of ~90 bp. These bands were not observed in normozoospermic donors. Microfluidid electrophoresis only identified the equivalent band of 150 bp. No correlation was observed between the intensity of the two bands and the level of SDF in males with SCI. Although DNase activity was present in the seminal plasma of both normozoospermic donors and men with SCI it did not digest cfDNA. cfDNA fragments were found to be hybridized all over the sperm genome. SCI patients with elevated sperm DNA fragmentation showed a 150 bp DNA band of cfDNA in the seminal plasma, which appeared resistant to DNase activity.

Mitigating the Effects of Oxidative Sperm DNA Damage.

Sperm DNA damage is correlated with reduced embryo development and increased miscarriage risk, reducing successful conception. Given its links with oxidative stress, antioxidants have been investigated as a potential treatment, yet results are conflicting. Importantly, individual antioxidants are not identical in composition, and some compounds may be more effective than others. We investigated the use of the polyphenol-rich, high-antioxidant-capacity fruit acai as a treatment for elevated sperm DNA fragmentation (>16%), measured by terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL). Following ≥ 74 days of treatment, we observed a significant decrease in sperm DNA fragmentation (−17.0% ± 2.5%) to 11.9 ± 1.7% (0–37%), with a 68.6% success rate (defined as post-treatment TUNEL < 16%). Post-treatment decreases in DNA fragmentation and success rates were not significantly impacted by low motility and/or concentration, or exceptionally high (> 25%) TUNEL. Treatment significantly reduced concentration in men with normal semen parameters, but 88% remained normal. Overall, successful treatment was not associated with age, semen parameters or TUNEL result at baseline. However, body mass index was significantly higher in nonresponders at baseline. This study provides evidence of a low-cost, effective treatment for elevated sperm DNA damage using acai.

326 - Clomiphene citrate and FSH treatment in men with elevated sperm DNA fragmentation index: Findings from a cross-sectional study

326 - Clomiphene citrate and FSH treatment in men with elevated sperm DNA fragmentation index: Findings from a cross-sectional study

The Use of Testicular Sperm for Intracytoplasmic Sperm Injection in Patients with High Sperm DNA Damage: A Systematic Review.

The advent of intracytoplasmic sperm injection (ICSI) has changed the human reproduction landscape by overcoming several limitations related to both male and female infertility factors. However, despite the development of new technologies, the live-birth rate with ICSI has not exceeded 30%. In order to improve assisted reproductive technology outcomes, advanced sperm function analysis have gained increased attention and the effects of sperm DNA fragmentation (SDF) on assisted reproduction success are being extensively studied. Utilizing ejaculated sperm with an elevated SDF has been found to result in poor ICSI outcomes. Furthermore, studies have reported that testicular sperm has lower SDF level, when compared to ejaculated sperm. This has led a number of clinicians world-wide to offer testicular sperm retrieval for ICSI in non-azoospermic males with high SDF. This practice has remained controversial due to lack of high quality evidence.

Does Use of Testicular Sperm Improve Outcomes in Nonazoospermic Couples With Previous IVF Failure Using Ejaculated Sperm? [OP02-2B

INTRODUCTION: Due to controversial evidence that testicular sperm is associated with lower sperm DNA fragmentation (SDF) and improved outcomes compared to ejaculated sperm (ES), this study evaluates intracytoplasmic sperm injection (ICSI) outcomes using testicular sperm in nonazoospermic couples with prior IVF failure using ES. METHODS: From Jan 2015-Aug 2018, nonazoospermic couples with ≥1 prior failed ART cycles using ES underwent testicular sperm extraction (TESE) for IVF/ICSI. Outcomes using TESE sperm were compared to the mean values of couples’ prior cycles using ES. Primary outcomes: clinical pregnancy and live birth rates (CPR &amp; LBR). Secondary outcomes: fertilization and blastocyst conversion. RESULTS: 64 couples were evaluated with an average of 2.5 prior failed ART cycles using ES. There were 76 total transfers including fresh/frozen transfers and supernumerary embryo transfers. Compared to prior failed cycles with ES, TESE sperm resulted equivalent fertilization rates (61.4% vs. 59.0%, p=0.66), but significantly improved blastocyst conversion rates (61.9% vs. 42.9%, p&lt;0.001) and blastocysts available for vitrification (1.6 vs. 0.6, p&lt;0.003). CPR and LBR were improved (42.1% and 35.5%, respectively), but not statistically compared to the prior failed cycles due to regression of the mean. A comparison of TESE-ICSI cycles in couples with ≥2 prior failed ART cycles using ES did not change the findings. CONCLUSION: In nonazoospermic couples with failed ART using ES, ICSI using TESE sperm may improve blastocyst development, number of embryos available for vitrification, CPRs, and LBRs. Testicular sperm may bypass adverse effects of elevated SDF from ES and improve pregnancy outcomes.

PD25-01 DOES THE USE OF TESTICULAR SPERM IMPROVE OUTCOMES IN NON-AZOOSPERMIC COUPLES WITH PREVIOUS IN-VITRO FERTILIZATION (IVF) FAILURE USING EJACULATED SPERM?

You have accessJournal of UrologyInfertility: Epidemiology & Evaluation II (PD25)1 Apr 2020PD25-01 DOES THE USE OF TESTICULAR SPERM IMPROVE OUTCOMES IN NON-AZOOSPERMIC COUPLES WITH PREVIOUS IN-VITRO FERTILIZATION (IVF) FAILURE USING EJACULATED SPERM? M. Blake Evans, Jessica Marinaro*, Allison Eubanks, Kate Devine, Micah Hill, Alan DeCherney, Russell Hayden, Paul Shin, and Cigdem Tanrikut M. Blake EvansM. Blake Evans More articles by this author , Jessica Marinaro*Jessica Marinaro* More articles by this author , Allison EubanksAllison Eubanks More articles by this author , Kate DevineKate Devine More articles by this author , Micah HillMicah Hill More articles by this author , Alan DeCherneyAlan DeCherney More articles by this author , Russell HaydenRussell Hayden More articles by this author , Paul ShinPaul Shin More articles by this author , and Cigdem TanrikutCigdem Tanrikut More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000000882.01AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: The use of testicular sperm for prior in-vitro fertilization (IVF) failure has been shown to have higher clinical pregnancy rates (CPR), live birth rates (LBR), and reduced spontaneous abortion (SAB) rates compared to ejaculated sperm (ES). However, some argue that this treatment is an invasive surgical procedure without clear benefit. This study evaluates the outcomes of IVF-intracytoplasmic sperm injection (ICSI) cycles using testicular sperm in non-azoospermic couples with prior IVF failure(s) using ES. METHODS: From 1/2015 to 10/2019, 116 non-azoospermic couples with ≥1 prior IVF cycle using ES underwent testicular sperm extraction (TESE) for IVF/ICSI. Failed cycles with ES were defined as those not progressing to clinical pregnancy. Outcomes using TESE sperm were compared to the mean values of couples’ prior cycles using ES. Primary outcomes included clinical pregnancy rates (CPR) and live birth rates (LBR). Secondary outcomes included fertilization and blastocyst conversion. RESULTS: Among 116 couples, the average number of prior failed IVF cycles using ES was 2.3 (range: 1-8). A total of 140 IVF/ICSI cycles were performed using TESE sperm (116 fresh TESE sperm, 24 frozen/thawed TESE sperm). Of these, there were 71 fresh blastocyst transfers, 31 frozen blastocyst transfers (FET), and 38 cycles without transfer. There were also 9 FETs using supernumerary embryos, for a total of 111 embryo transfers. Compared to ES, IVF/ICSI using TESE sperm significantly improved blastocyst development (p=0.003), blastocyst conversion (p=0.003) and number of embryos available for vitrification (p=0.003). A subset of men (n= 66) had a sperm DNA fragmentation (SDF) assessment of ES. Mean SDF was 45% (7- 90%). Of these 66 men, 55 (83%) had an SDF >25%. The outcomes for patients with elevated SDF did not differ significantly from the overall cohort. CONCLUSIONS: In non-azoospermic couples with prior failed IVF cycles using ES, IVF/ICSI using TESE sperm may improve embryo development, CPR and LBR. Testicular sperm may avoid the adverse effects of elevated SDF from ES and improve pregnancy outcomes in some patients. Further randomized studies are needed to determine if such a benefit exists. Source of Funding: None © 2020 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 203Issue Supplement 4April 2020Page: e538-e538 Advertisement Copyright & Permissions© 2020 by American Urological Association Education and Research, Inc.MetricsAuthor Information M. Blake Evans More articles by this author Jessica Marinaro* More articles by this author Allison Eubanks More articles by this author Kate Devine More articles by this author Micah Hill More articles by this author Alan DeCherney More articles by this author Russell Hayden More articles by this author Paul Shin More articles by this author Cigdem Tanrikut More articles by this author Expand All Advertisement PDF downloadLoading ...

MICROFLUIDIC SPERM SEPARATION DEVICE DRAMATICALLY LOWERS DFI

Elevated sperm DNA Fragmentation Index(DFI) has been shown to be associated with decreased pregnancy rate, implantation rates, and increased miscarriage rates(1). Increasing DFI has been correlated with abnormal semen parameters, most notably morphology and linear progression(2). Standard semen processing which includes centrifugation may increase reactive oxygen species(ROS) and increased DNA fragmentation in sperm(3). Microfluidic based sperm separation allows for the selection of highly motile sperm with improved morphology and has not been shown to increase ROS.