Sperm DNA Fragmentation Index Research Articles

P-053 Sperm chromatin integrity in men from infertile couples with teratozoospermia and normozoospermia

Abstract Study question What is the relationship between sperm morphology and sperm chromatin integrity (SCI) in patients with normal sperm concentration (SC) and motility (SM)? Summary answer SCI violations in form of sperm DNA fragmentation (SDF) and incomplete protamination (IP) are more common in teratozoospermia, even when SC and SM are normal What is known already Deterioration of sperm chromatin integrity in the form of SDF and IP are the significant factors affecting male reproductive potential (WHO, 2021). Study design, size, duration This multicenter retrospective cross-sectional study included 479 men aged 18-45 years who were observed for infertility for more than 1 year. Participants/materials, setting, methods Only men: 1) with sperm concentration and motility that were not lower than the 5% centile according to WHO-2021 (16 million/ml and 30%, respectively), 2) without any inflammatory processes in genitourinary organs, 3) with obtained tests for SCI were included in the study. The men were divided into 3 groups (gr.) depending on the percentage of morphologically abnormal forms: I - 86-95% (n = 229), II - 96-99% (n = 228), III - 100% (n = 22). Main results and the role of chance The mean SDF in groups I, II, III were 9.0% (6-15), 12.0% (8-16) and 16.5% (13-20) (pI-II = 0.009; pII-III = 0.002; pI-III = 0.001), respectively. SDF index (Halosperm® Assay) in groups I, II, and III were 1.3 (1.2-1.5), 1.4 (1.3-1.5), and 1.5 (1.4-1.7), respectively (pI-II = 0.004; pII-III = 0.001; pI-III = 0.009). In 23% (I), 27% (II), and 55% of cases (III) the SDF value exceeded the threshold value (>15%) (pI-II > 0.05; pII-III <0.02; pI-III <0.005). Histone-positive sperm were revealed in 17% (12-23), 19.5% (13-27), and 31% (19-42) of cases in groups I, II, and III, respectively (pI-II > 0.05; pII-III = 0.014; pI-III = 0.02); The 30% of SDF threshold value was exceeded in groups I, II, and III, in 12%, 20%, and 52% of cases (pI-II < 0.05; pII-III < 0.005; pI-III < 0.001). Limitations, reasons for caution The reference values of SDF and the proportion of histone-positive sperm have not yet been determined properly, even in WHO 2021, therefore, the prevalence of SCI abnormalities in various spermiological diseases needs to be clarified in the future. Wider implications of the findings SCI and IP can be applied more widely in the investigation of men from infertile couples, especially, men with teratozoospermia. It should also be considered in men from infertile couples with normal semen parameters. Trial registration number Not applicable

P-082 Microfluidic-based device selects sperm with less DNA damage and higher motility, what else?

Abstract Study question Does the microfluidic-based sperm selection device ZyMōt improve sperm parameters and other laboratory key performance indicator (KPI) values compared to the conventional swim-up method? Summary answer The microfluidic-based sperm selection device ZyMōt selects sperm with lower DNA fragmentation and higher motility than conventional swim-up method in intracytoplasmic sperm injection (ICSI) cycles. What is known already Elevated levels of sperm DNA fragmentation (SDF) in semen samples have been associated with poor embryo development and low pregnancy rates. SDF refers to breaks in the sperm's genetic material, mainly due to defects during spermatogenesis and other factors such as reactive oxygen species that are favored by centrifugation. Conventional sperm selection methods, by integrating centrifugation into their protocols, have become ineffective in selecting sperm with low SDF. In order to solve this problem and improve reproductive outcomes, microfluidic-based devices such as ZyMōt have been designed to avoid centrifugation and select spermatozoa with low SDF. Study design, size, duration Prospective, experimental, single-center study conducted from June to December 2021. A total of 14 couples with ≥ 10 retrieved oocytes were recruited for an intra-patient comparison. Semen sample was split and processed for ICSI by the conventional swim-up method or by the microfluidic-based device ZyMōt. Each fraction was used to fertilize half of the total number of oocytes retrieved. SDF index, semen parameters, useful blastocyst rate, fertilization rate and morphokinetic variables were observed. Participants/materials, setting, methods Oocytes retrieved were from own (n = 96) and donation cycles (n = 93). From each patient, the cohort of oocytes was divided into two groups: 1) inseminated with spermatozoa selected by swim-up and 2) inseminated with spermatozoa selected by ZyMōt 850 µL device. Embryo evaluation and development were then followed by time-lapse monitoring using EmbryoScope. Sperm chromatin dispersion (SCD) assay was used to measure SDF, analyzed by ImageJ. Each treatment followed routine protocol established in the clinical practice. Main results and the role of chance SDF index was significantly lower in ZyMōt group in comparison with swim-up group (10% vs 20%), indicating a better selection by the ZyMōt of sperm with less DNA breaks. Additionally, ZyMōt group also presented a significantly greater number of spermatozoa with progressive motility (96.9% vs 95.4%). In contrast, useful blastocyst rate showed a slightly, but not significantly, increment in ZyMōt group compared to swim-up group (53.2% vs 46.6%). No significant differences in fertilization rate or sperm recovery rate were observed between groups. Regarding morphokinetic parameters, timing variables from first cell division to blastocyst stage (t2-tB) showed no significant correlation with ZyMōt group contrasted with swim-up. Blastocysts were evaluated and a value was assigned with respect to their quality (A to D). There was a higher number of embryos with A grade in ZyMōt group and a higher number of embryos with D grade in swim-up group. The annotated variables were assessed using paired t-test and P value <0.05 was considered statistically significant. Limitations, reasons for caution The impact on reproductive outcomes may vary depending on whether the breakage is single- or double-stranded, however, SCD is not able to distinguish between them. These, together with the oocyte's ability to repair sperm damage, lead us to the explanation for the non-significance on embryo quality. Wider implications of the findings This study provides further insights into the use of ZyMōt for the selection of low SDF spermatozoa in a simple manner. Its use could be indicated in patients with a high SDF value in their semen samples. Benefits on reproductive outcome will be confirmed with larger sample size. Trial registration number 2102-VLC-007-MD

P-549 Paraclinical and spermogram evaluation of infertile males with balanced chromosomal rearrangements referred to Royan institute from 2009-2021

Abstract Study question Whether balanced chromosomal rearrangements are associated with altered semen parameters/increased DNA Fragmentation Index (DFI), are the common breakpoints related to male infertility in patients ? Summary answer Balanced chromosomal rearrangements is associated with abnormal semen parameters and increased DFI in carriers. Chromosomes 9,1,3 showed the highest incidence of chromosomal rearrangement What is known already Among many factors, balanced chromosomal rearrangements are thought to be one of the major genetic factors influencing male infertility which can cause disorders related to spermogram, DNA fragmentation, increased risk of miscarriage and ART failure. Finding common chromosomal breakpoints in infertile men carrying balanced chromosomal translocations is significant since these patients usually possess normal clinical features. Numerous studies have been performed on the various chromosome translocations. However, there has not been any report on the common breakpoints in balanced chromosomal abnormalities related to male infertility in Iran. Study design, size, duration This cross-sectional study performed on 25000 patients’ files admitted to the Royan institute during the period 2009- 2021.Infertile men carrying balanced chromosomal rearrangements were included as case group. Sperm parameters, DFI and common chromosomal break points were evaluated Participants/materials, setting, methods 25000 files of patients were reviewed. 252 infertile men carrying balanced chromosomal rearrangements and 252 men with normal karyotype were included (case and control group, respectively). Sperm concentration, motility (CASA), sperm morphology (Papanicolaou Staining),DFI (SCSA) and cytogenetic analysis of peripheral blood lymphocytes (G-banding) were evaluated. Numerical chromosomal abnormalities, deletions, marker chromosomes, alcohol or drug addiction, smoking, chemotherapy and radiation, varicocele and exposure to environmental pollution were excluded. The common chromosomal breakpoints in translocations were investigated. Main results and the role of chance Sperm concentration, total motility and normal morphology were significantly lower in balanced chromosomal rearrangements carriers compared to control group(p < 0.05). DFI in patient was significantly higher in comparison to control group(p < 0.05). Among different kind of chromosomal rearrangement carriers’, sperm concentration and total motility were lower in Robertsonian translocation carriers comparing to the Reciprocal translocation and inversion ones. The reciprocal translocation and inversion carriers did not differ regarding to sperm concentration and total motility. Meanwhile, normal morphology was lower in the translocation compared to inversion carriers. No difference was observed in DFI among carriers of balanced chromosomal rearrangements. Among case group, inversion accounted for the largest share, followed by Reciprocal, Robertsonian translocation and insertion. Group C had the highest frequency in reciprocal translocation, inversion and insertion among the chromosomal groups. The highest balanced translocation prevalence was related to three chromosomes 9,3, and1. 9q12,1p36,3p11 were the most frequent breakpoints among them. Chromosome 9 had the largest contribution to inversions,22 to reciprocal translocations and14 to Robertsonian translocations. In the Robertsonian translocation, group D showed the highest frequency. Interestingly, patients carrying normal variations such as pericentric inversion of chromosome 9showed a significant alteration in sperm parameters, therefore normal variations showed a possible relationship to infertility. Limitations, reasons for caution In order to achieve stronger statistical results on DFI, a larger sample size would be more helpful. Wider implications of the findings The results may be useful to identify the most common breakpoints and consequently genes related to male infertility and can suggest appropriate molecular mechanism for their infertility. Trial registration number not applicable

O-132 Is there an association between sDF index and serum testosterone in human follicle stimulating hormone alfa (r-hFSH alfa) treated males with idiopathic infertility?

Abstract Study question Is there correlation between testosterone serum levels and sperm DNA fragmentation (sDF) index before and after follicle-stimulating hormone (FSH) administration in male idiopathic infertility? Summary answer Correlation between testosterone and sDF was highlighted for the first time in FSH-treated men. Significant amelioration of conventional semen parameters and sDF index were confirmed. What is known already Testicular overstimulation is the yearned therapeutic goal when exogenous FSH is empirically administered in male idiopathic infertility. Although a robust physiological rationale theoretically supports the FSH use in male idiopathic infertility, useful markers to evaluate its efficacy are still far to be identified. Whether pregnancy outcomes remain the expected valuable endpoint for couple infertility treatment, the identification of reliable, and possibly early, markers of therapeutic response to FSH in males is mandatory. Randomized controlled clinical trials (RCTs) on this topic focused, as main outcomes of FSH treatment effectiveness, on testicular seminiferous component, not considering involvement of testosterone-secreting compartment. Study design, size, duration A retrospective post-hoc analysis of raw data extracted from two RCTs in which 148 idiopathic infertile men were treated with FSH. Participants/materials, setting, methods A retrospective post-hoc re-analysis was performed on raw data of RCTs in which idiopathic infertile men were treated with FSH and both testosterone serum levels and sDF were reported among primary and/or secondary endpoints. Additional data regarding couple infertility history, age, anthropometric variables, FSH treatment scheme and semen variables were included in a single dataset. Data were evaluated by logistic regression analyses. Main results and the role of chance Two RCTs were included accounting for 148 patients (median age 37, 25-52 years). After three months of FSH administration, a significant increase was observed in FSH levels (p < 0.001), inhibin B (p = 0.012), sperm concentration (p = 0.003), total sperm number (p = 0.021), progressive motility (p < 0.001) and normal sperm morphology (p < 0.001). Moreover, an overall sDF index reduction was confirmed after treatment (19.3±9.4 versus 16.4±6.8, p = 0.002). SDF was found significantly inversely related to sperm concentration both at baseline and after FSH treatment (Rho -0.325, p < 0.001 and Rho -0.316, p = 0.001, respectively). Interestingly, sDF index after treatment showed a significant inverse correlation with testosterone serum levels (Rho -0.327, p = 0.002). Multivariate stepwise linear regression analyses using sDF index as dependent variable identified testosterone as a predictor for sDF index change (p = 0.005). Similarly, logistic regression analysis highlighted testosterone and SHBG levels as predictive of sDF reduction after FSH administration (p = 0.043 and p = 0.005, respectively). Combining raw data of published RCTs investigating FSH administration to idiopathic infertile men, a significant amelioration of conventional semen parameters together with a reduction in sDF were confirmed. Intriguingly, a potential correlation between serum testosterone and sDF was highlighted for the first time. Limitations, reasons for caution Since the analysis was performed on raw data obtained in previous RCTs, heterogeneity among patients enrolled and evaluated should be considered interpreting these results. Moreover, a real and accurate evaluation of intratesticular testosterone levels, rather than serum levels should be more informative to the objective of the study. Wider implications of the findings The highlighted significant correlation between testosterone serum levels and sDF is opening a completely unexplored field in the possibility to identify early predictors of FSH therapy response in male idiopathic infertility. Trial registration number Not applicable

P-430 Sperm genomic integrity, telomere dynamics and gene expression in recurrent implantation failure

Abstract Study question Does sperm affect the implantation and early embryogenesis? Summary answer Deranged sperm genomic integrity, limited repair mechanisms, aberrant gene expression have the potential to be transmitted to developing embryo and affect implantation and embryonic development. What is known already The terminally differentiated, transcriptionally quiscent sperm cells are vulnerable to a host of factors which might affect its function. The spermatozoa with truncated repair mechanisms have the potential to fertilize the oocyte, but eventually result in high rates of pre and post implantation losses. Oxidative stress, deranged genomic integrity, aberrant gene expression in the sperm cell has the potential to affect fertilisation potential, implantation and embryonic viability. This has been seen to be correlated with decreased fertilisation, clevage rates and blastocyst development and thus impaired implantation rates. Study design, size, duration A case control study of 75 men from infertile couples who had recurrent implantation failure (RIF) in IVF cycles and 75 fertile controls at AIIMS, New Delhi, India. Study duration was 2 years. Participants/materials, setting, methods Semen samples from men from couples who had RIF were analyzed as per WHO 2010. Sperm reactive oxygen species (ROS) and DNA fragmentation index (DFI) was assessed by chemiluminiscence and sperm chromatin structure assay (SCSA) respectively. Relative sperm telomere length was evaluated from sperm DNA by q-PCR analysis. The expression of genes pertinent for early embryonic development was done by qPCR. The relative quantification was done after normalizing with GAPDH and β-actin by 2-ΔΔCt method. Main results and the role of chance Seminal ROS levels (RLU/sec/million sperm) were seen to be significantly higher [57.75 (10.1-1186.9)] in cases with respect to controls [16.7(1.15-53.9)] (p < 0.001***). The mean DFI of men undergoing ART was significantly higher (37.7 ± 5.7) vs 23.2 ± 4.6%; p < 0.001***) in cases as compared to controls. ROS and DFI levels correlated negatively with sperm concentration and progressive motility (p < 0.001***). We analyzed the expression of FOXG1, SOX3, STAT4, RPS6, RBM9, RPL10A, RPS17, RPL29, WNT5A, HSP90, TOMM7, EIF5A genes. The expression of SOX3, RBM9, WNT5A, HSP90, TOMM7, and EIF5A showed a significant difference from control levels. The relative sperm telomere length was found to be significantly lower in RIF patients as compared to controls (p < 0.001***). Limitations, reasons for caution The current study was a case control study and lacked randomization and also is limited by low sample size. There is a lack of stratification in the enrollment of participants due to difference in specific risk of occurrence and varied clinical history. Wider implications of the findings The analysis of spermatozoal gene expression is important for understanding the sperm differentiation, fertilization and early embryonic events. Correlation with oxidative stress, genomic integrity and telomere length may help in regulationof gene expression. It may help in establishing sperm gene expression as a potential biomarker. Trial registration number Not applicable

P-070 Comparison of Microfluidic Sperm Sorting (MFSS) versus Physiological Intracytoplasmic Sperm Injection (PICSI) versus Density Gradient versus Swim Up in high DNA fragmentation index sperm samples

Abstract Study question To evaluate the effectiveness of using Microfluidic Sperm Sorting (MFSS) over other technique in patient with high DNA fragmentation index (DFI) sperm samples? Summary answer Microfluidic is not only correlated with better DNA integrity but also with the better reproductive outcome. What is known already DNA damage is unrecognisable in living sperm prior to insemination and an increased sperm DNA fragmentation index has been associated with lower fertilization rates, impaired embryo development and reduced pregnancy rates. Standard semen processing techniques are associated with centrifugation, which may induce reactive oxygen species and DNA damage. In strategies to minimize sperm DNA fragmentation, Physiological ICSI can relatively reduce sperm DNA fragmentation by 67.9% (Parmegiani et al., 2010) while new technique Microfluidic sperm sorter technique also demonstrate sperm selection with significantly reduced DNA damage Study design, size, duration A prospective randomised study was conducted from 1st January 2019 to 1st December 2021. Four hundred patients were randomised by computer generated list and divided into 4 groups. Group A (n = 100) , in which sperm were processed by microfluidic sperm sorter (MFSS) while in group B (n = 100), sperm were selected by Physiological Intracytoplasmic Sperm Injection (PICSI), Group C (n = 100), sperm were processed by density gradient and Group D (n = 100), sperm were processed by swim-up technique. Participants/materials, setting, methods The study period included all normozoospermic patients with high DNA fragmentation index (>25%) while oligospermic, asthenozoospermic samples, patients with poor ovarian reserve and advanced age were excluded from the study. All A grade embryos were vitrified and transferred in frozen embryo replacement cycle. All the 3 groups were compared on the basis of fertilisation rate, day 3 grade A embryo development rate, clinical pregnancy rate and miscarriage rate. Main results and the role of chance Cycle characteristics (female age, length of stimulation, gonadotrophin dose, number of oocytes and number of transferred embryos) were similar among all 4 groups. Among 4 groups, There was a statistically significant increase observed in Group A (Microfluidic Sperm Sorting), day 3 grade A embryo development rate (60% vs. 42% vs. 38% vs. 40%, p-0.016) and clinical pregnancy rate (62% vs. 46% vs. 41% vs 43%, p-0.049), while no statistical significant difference observed in fertilisation rate (82% vs. 78% vs. 76% vs. 78%, p-0.80) comparing group B, group C and group D. But significantly higher miscarriage rate (12% vs. 11% vs. 25%, 12%) observed in density gradient (Group C) technique. Limitations, reasons for caution Larger randomised control studies are needed to strengthen these results. Wider implications of the findings We have demonstrated that sperm sorted by microfluidic helps in selection of sperm with better DNA integrity over Physiological ICSI, Density Gradient and Swim-up techniues. Using it in routine practice can help in reducing the negative effect of reactive oxygen species and thus improve pregnancy rate and live birth rate. Trial registration number MCDH/2019/35

P-046 Effect of different sperm chromatin dispersion type on IVF/ICSI outcome and offspring profile

Abstract Study question Whether the percentage of different sperm chromatin dispersion type are associated with the IVF/ICSI outcome and offspring profile? Summary answer Percentage of different sperm chromatin dispersion type are significantly associated with the embryo cleavage rate, embryo quality, live birth rate and offspring gender. What is known already The sperm DNA fragmentation index (DFI) is increasingly recognized as a key sperm functional parameter for assessing male fertility. Sperm chromatin dispersion (SCD) test is widely used in clinical andrology lab to measure DFI, and according to the size of sperm halo it divides sperm into three categories: big halo sperm, middle halo sperm, small or no halo sperm. Previous study reported that big halo sperm are significantly associated with embryo quality and pregnancy rate. The average halo size of Y-chromosome bearing sperm are significantly smaller than X - chromosome bearing sperm. Study design, size, duration This is a multi-center retrospective study recruited 200 couples underwent IVF/ICSI treatment in Nanfang Hospital and Guangdong Women's and Children's Hospital from May 2018 to May 2019. Participants/materials, setting, methods Patients meet the followed inclusion criteria were recruited: (1) The male partners received sperm chromatin dispersion test; (2) The female partners have normal ovarian reserve (female age < 35 years old, FSH ≤10 IU/L and AFC ≥ 5); (3) Couples received fresh single embryo transfer. The association between sperm chromatin dispersion type and IVF/ICSI outcome and the offspring profile were retrospectively analyzed. Main results and the role of chance Regression analysis showed that the embryo cleavage rate is positively associated with the percentage of sperm with big halo (Adjusted ?????(95% CI) 0.09 (0.01, 0.17), P<0.05), but negatively associated with the percentage of sperm with middle halo (Adjusted ????? (95% CI)- 0.22 (-0.35, -0.09), P<0.001). The good embryo rate is negatively associated with the percentage of sperm with middle halo (Adjusted ????? (95% CI) -0.60 (-1.04, -0.16), P<0.05). Live birth is significantly associated with the percentage of sperm with small halo (Adjusted OR (95% CI) 0.89 (0.80, 1.00), P<0.05). Offspring gender is significantly associated with the percentage of sperm with big halo (Adjusted OR (95% CI) 0.95 (0.91, 0.99), P<0.05) and the percentage of sperm with middle halo (Adjusted OR (95% CI) 1.09 (1.02, 1.17), P<0.05). The area under the receiver operator characteristic (ROC) curve (AUC) of the big halo sperm percentage to predict male offspring is 0.68 (95%CI (0.54, 0.81), P < 0.05). The AUC of middle halo sperm percentage to predict female offspring is 0.66 (95%CI (0.52, 0.80), P < 0.05). Limitations, reasons for caution This is a retrospective study which may have the bias caused by the nature of this type of study. The sample size is relatively small. Wider implications of the findings The percentage of big halo sperm are significantly associated with embryo cleavage rate and male offspring. The percentage of middle halo sperm are significantly associated with embryo cleavage rate, lower good embryo rate and female offspring. The percentage of small halo sperm is significantly associated with live birth. Trial registration number 82101683

Sperm DNA Fragmentation Index: A Comparison Study of Success Rates among Natural, Intrauterine Insemination (IUI) and In Vitro Fertilization (IVF)-Intra Cytoplasmic Sperm Injection (ICSI) Pregnancy Programs

Background: Some studies have reported a relationship between the sperm DNA fragmentation index (DFI) and the rate of fertilization and pregnancy. This study was designed to assess the mean sperm DNA fragmentation in pregnancies that occur in infertile couples, whether in natural pregnancy, intrauterine insemination (IUI), and in vitro fertilization (IVF) – intra cytoplasmic injection (ICSI).
 Methods: This research is an observational analytic study with a cross-sectional design. The sample taken in this study were infertile patients that underwent natural pregnancy or IUI or IVF-ICSI at Yasmin Infertility Clinic of Dr. Ciptomangunkusumo Hospital period 2018-2020 with a consecutive sampling technique. The research data was processed and analyzed by the Mann-Whitney test using the SPSS application.
 Results: The mean DFI of sperm in men with infertile couples who successfully conceived naturally was 10.7% (mild), while IUI was 20.4% (moderate), and IVF-ICSI was 30.5% (poor). The mean DFI in semen samples of men from infertile couples who underwent a natural program was significantly lower in those who successfully conceived compared to those who did not. Similar results were also shown in the IUI and IVF-ICSI programs, which showed a significantly lower DFI compared to non-pregnant women.
 Conclusion: DFI can be applied as a marker for selecting the type of pregnancy program in infertility management.

Влияние гиперкапнической гипоксии на репродуктивную и копулятивную функции мужчин в комплексном лечении хронического абактериального простатита.

Introduction. The relationship between chronic abacterial prostatitis/chronic pelvic pain syndrome (CP/CPPS) and male infertility is the subject of numerous studies, and therefore various methods for correcting pathospermia have been proposed. The main concept is that in one way or another it is necessary to create conditions for a more perfect penetration of drugs into the zone of the prostate-vesicular complex. In this regard, the search for new and pathogenetically substantiated methods of therapy for such a patient seems to be very relevant Aim of study. To evaluate the effect of hypercapnic hypoxia on reproductive and copulatory functions of men in the complex treatment of CP/CPPS. Materials and methods. The study involved 24 patients who applied to a urologist with complaints about the absence of conception in marriage for 1 year or more. The examination revealed pathospermia and symptoms of CP/CPPS in these patients. Assessment of the severity of symptoms of CP/CPPS was assessed using a questionnaire on the symptoms of chronic prostatitis using the NIH-CPSI questionnaire. In order to identify and monitor violations of the male copulatory function, ICF questionnaires created by O.B. Loran and A.S. Segal and the International Index of Erectile Function (MIEF-5). Reproductive function analysis was monitored using a semen analysis and sperm DNA fragmentation index test. The patients were divided into 2 groups of 12 people. Both groups received phenotypically directed pathogenetic and symptomatic therapy. For patients of the 2nd group, the complex treatment included breathing training procedures using the carbonic simulator, which ensures blood saturation with CO2, leading to the effect of hypercapnic hypoxia. This phenomenon contributes to angiogenesis and improvement of blood circulation in organs and systems. The effectiveness of the treatment was evaluated after 1 and 3 months. Results. According to the results of the NIH-CPSI questionnaire, 3 months after treatment, therapy in patients of group number 2 was more effective. According to the results of the MIEF 5 and ICF questionnaires in patients with CP/CPPS, a violation of erectile and ejaculatory function was revealed, and according to the results of the spermogram and the sperm DNA fragmentation test (IFD), pathospermia was noted, which was expressed in a decrease in the concentration of spermatozoa (group number 1 – 15.3 ± 1.6 million / ml, group number 2 – 16.8±2.2 million/ml), their progressive mobility (group number 1 – 24 ± 2.6%, and in group number 2 23.25 ± 2.9%.), normal morphology (group number 1 – 2.1 ± 0.17%, group number 2 – 2.5 ± 0.29%) and an increase in IFD (group number 1 – 17.9 ± 2.06%, group number 2 – 19.75± 1.9%). 3 months after treatment, the concentration of spermatozoa was 21.1±2.16 million/ml in group number 1, and 31.8±2.5 million/ml in group number 2. Progressive mobility in group number 1 – 31.4±2.2%, in group number 2 – 38.4±1.9%. Normal morphology in group number 1 is 3.1 ± 0.32%, in group number 2 – 4.6 ± 0.6%. IFD in group number 1 – 12.25 ± 0.6%, in group number 2 – 7.8 ± 1.3%. Conclusion. Patients of the group number 2, who trained on the breathing device «carbonic», achieved a more significant clinical effect from the therapy. Hypercapnic hypoxia, in the complex treatment of CP/CPPS, can contribute not only to the relief of pain and dysuric symptoms, but also to improve both the copulative and reproductive functions of men.

Efficacy of combination of multinutrients in reduction of sperm DNA fragmentation

High Sperm DNA fragmentation can cause decreased fertility & implantation rate or can cause Recurrent implantation failure and if fetus born then can lead to Genetic abnormality in the fetus such as Down Syndrome, Edward syndrome or Patau’s syndrome. The objective of the study is to combine the peripheral data of the patients to evaluate the safety and efficacy of Combination tablet in reduction of Sperm DNA fragmentation index. The result shows that the factors affecting the DFI .The Institutional Ethics Committee approved the study procedure (Ref No: SRMC/RP/4505).

Efficacy of combination of multinutrients in reduction of sperm DNA fragmentation

High Sperm DNA fragmentation can cause decreased fertility & implantation rate or can cause Recurrent implantation failure and if fetus born then can lead to Genetic abnormality in the fetus such as Down Syndrome, Edward syndrome or Patau’s syndrome. The objective of the study is to combine the peripheral data of the patients to evaluate the safety and efficacy of Combination tablet in reduction of Sperm DNA fragmentation index. The result shows that the factors affecting the DFI .The Institutional Ethics Committee approved the study procedure (Ref No: SRMC/RP/4505).

Comparison of zeta potential and physiological intracytoplasmic sperm injection in obtaining sperms with a lower DNA fragmentation index: A cross-sectional study.

BackgroundThe sperm DNA fragmentation index (DFI) is one of the men's reproductive health criteria that affects assisted reproductive technique outcomes. Efforts in obtaining high-quality mature sperms seem to be necessary. Advanced sperm selection techniques (including physiological intracytoplasmic sperm injection [PICSI], zeta potential, microfluidic, etc.) have gained popularity in this regard.ObjectiveThe study aimed to compare the efficacy of zeta potential and PICSI sperm selection in obtaining sperms with better DNA integrity.Materials and MethodsIn this cross-sectional study, 48 couples were enrolled where the male partner had increased sperm DFI in his ejaculated sample and the female was in normal reproductive health. For each male partner, the semen sample was processed with zeta potential and PICSI techniques, then the sperm DFI of neat semen was compared to zeta and PICSI samples by the sperm chromatin dispersion test.ResultsData showed that both the zeta potential and PICSI technique decreased sperm DFI in comparison with the neat semen sample (p 0.001 for both). In addition, there was a statistically significant difference in sperm DFI between the PICSI and zeta potential samples (p 0.01).ConclusionThe current study showed that both zeta potential and PICSI could result in sperm with a lower DFI. However, PICSI seems to be superior to zeta potential in this regard.

The role of sperm DNA integrity in couples with recurrent implantation failure following IVF treatment.

There is increasing evidence from the literature that indicates the association between impaired sperm DNA integrity and male infertility. However, the data is insufficient regarding recurrent implantation failure (RIF) and sperm DNA damage. This study aimed to investigate the association between sperm DNA fragmentation and RIF cases. Basic semen parameters and sperm DNA fragmentation index (DFI) of men whose partner was suffering from RIF were compared with men whose partner was diagnosed with unexplained infertility (UEI) but had clinical pregnancies following IVF treatment. A retrospective analysis from a large-volume IVF center has been performed, and a total of 197 couples underwent analysis. Two groups were formed, couples with RIF and couples diagnosed with UEI but had clinical pregnancies (controls) following IVF cycles. The mean number of cycles showed significant differences between the groups. However, no statistical difference was observed between RIF and the control group regarding patient characteristics, semen parameters, and sperm DNA fragmentation index (DFI). Also, no statistically significant correlation was found between sperm DFI and clinical pregnancies in the unexplained infertility cohort. Our results show that sperm DNA fragmentation may not be an important contributing factor to RIF cases.

The Effect of Sperm DNA Fragmentation on In Vitro Fertilization Outcomes for Women With Polycystic Ovary Syndrome

BackgroundPolycystic ovary syndrome (PCOS) is a prevalent endocrine disease in reproductive women associated with poor pregnancy outcomes. In modern society, people pay more attention to the female factor, but it is uncertain whether sperm quality is another factor affecting pregnancy outcomes of patients with PCOS.MethodThe effect of sperm DNA fragmentation (SDF) on oocyte fertilization, embryonic development, and pregnancy outcomes for patients with PCOS who underwent in vitro fertilization (IVF) treatment was studied. A total of 141 PCOS patients and 332 control patients undergoing IVF treatment were recruited from January 2017 to December 2019. All female patients were designated into two groups according to the Rotterdam criteria. Each group was divided into two sets, DNA fragmentation index (DFI) ≤15% and DFI > 15%, on the basis of sperm DFI.ResultThere were no differences in basic clinical characteristics between couples with a sperm DFI ≤ 15% or DFI > 15%. For control patients, no differences were observed in IVF outcomes. However, for PCOS patients, although there were no differences in the fertilization (60.4% vs. 59.9%, p = 0.831), high-quality embryo (68.5% vs. 67.9% p = 0.832), clinical pregnancy (78.4% vs. 66.7%, p = 0.148), and abortion (12.5% vs. 11.5%, p = 1.000) rates, a significantly lower high-quality blastocyst formation rate (26.3% vs. 16.3%, p = 0.023) was observed for couples with a sperm DFI > 15% compared with a sperm DFI ≤ 15%.ConclusionFor PCOS patients undergoing IVF, oocytes fertilized using sperm with higher DFI led to a lower high-quality blastocyst formation rate but had no influence on fertilization, high-quality embryo, clinical pregnancy, and miscarriage rates.

The FSH administration to men with idiopathic infertility improves sperm DNA fragmentation index together with testosterone serum levels increase

Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

PD36-02 HISTOLOGICAL EVIDENCE OF SPERMATOGENIC FAILURE IN MEN WITH HIGH SPERM DNA FRAGMENTATION INDEX

You have accessJournal of UrologyCME1 May 2022PD36-02 HISTOLOGICAL EVIDENCE OF SPERMATOGENIC FAILURE IN MEN WITH HIGH SPERM DNA FRAGMENTATION INDEX Taras Shatylko, Inessa Telezhnikova, Safar Gamidov, Alikhan Tambiev, Alina Popova, and Vlada Kometova Taras ShatylkoTaras Shatylko More articles by this author , Inessa TelezhnikovaInessa Telezhnikova More articles by this author , Safar GamidovSafar Gamidov More articles by this author , Alikhan TambievAlikhan Tambiev More articles by this author , Alina PopovaAlina Popova More articles by this author , and Vlada KometovaVlada Kometova More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002594.02AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Recurrent pregnancy loss and IVF failures may be attributed, among other factors, to sperm DNA fragmentation (SDF). Many conditions and lifestyle factors potentially cause sperm DNA damage, but it is currently unknown where exactly this damage occurs – in testis, epididymis or other portion of seminal tract. Various studies have demonstrated lower SDF index in testicular sperm when compared to ejaculated sperm. Intracytoplasmic testicular sperm injection (Testi-ICSI) is a controversial technique using surgically retrieved sperm from non-azoospermic men to improve IVF outcomes. We sought to evaluate histopathology of seminiferous tubules obtained during sperm retrieval for Testi-ICSI. METHODS: This is an ongoing prospective study. The inclusion criteria are: recurrent pregnancy loss or at least 2 IVF failures, elevated DNA fragmentation index (TUNEL >15%). The exclusion criteria are: known female factor causing pregnancy loss or IVF failures, abnormal karyotype, oligozoospermia (concentration <15 × 106; total sperm count <39 × 106), treatable causes of increased SDF index, history of hormone therapy. Eighteen patients underwent testicular sperm extraction (TESE) for Testi-ICSI. A portion of extracted tissue was sent to pathology. Specimens taken from 5 patients undergoing TESE for obstructive azoospermia (duration of obstruction less than 5 years) were used as controls. Bergmann-Kliesch score (BKS) was used to assess seminiferous tubules histology. Fisher’s exact test and Mann-Whitney test were used as a part of statistical analysis. RESULTS: Median DNA fragmentation index for ejaculated sperm was 26% (interquartile range [IQR]: 23-30). Median percentage of seminiferous tubules containing elongated spermatids was 74% (IQR: 61-89), while in control specimens it was 92% (IQR: 90-95), which was a statistically significant difference (p=0.0251). Median BKS in Testi-ICSI specimens was 7 (IQR: 6-9), which was significantly lower than in control specimens (median: 9; IQR: 9-10; p=0.0271). Hypospermatogenesis was a dominant histological pattern in 10 of 18 Testi-ICSI specimens (55.5%), while all control specimens had features of normal spermatogenesis (p=0.0457). CONCLUSIONS: High prevalence of hypospermatogenesis in non-oligozoospermic patients was an unexpected finding. In theory, Testi-ICSI should work better in cases of post-testicular sperm DNA damage. However, we have found histological evidence of spermatogenic failure in some patients, which hints at probable testicular origin of SDF, which is yet to be correlated with Testi-ICSI outcomes. Source of Funding: N/A © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e633 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Taras Shatylko More articles by this author Inessa Telezhnikova More articles by this author Safar Gamidov More articles by this author Alikhan Tambiev More articles by this author Alina Popova More articles by this author Vlada Kometova More articles by this author Expand All Advertisement PDF DownloadLoading ...

Prenatal exposure to airborne polychlorinated biphenyl congeners and male reproductive health.

Is fetal exposure to lower-chlorinated polychlorinated biphenyls (LC-PCBs) in indoor air of private homes built with PCB-containing materials associated with semen characteristics and testicular volume in adult men? We observed only marginal and inconsistent associations between maternal exposure to PCBs in indoor air and semen quality, testicular size and reproductive hormones in the adult offspring. Recent studies have shown LC-PCBs to exhibit endocrine-disrupting properties and increase the risk of cryptorchidism. Although exposure to LC-PCBs in indoor air is relatively common, the long-term impact of prenatal exposure on male reproductive health has not yet been investigated. In this cohort study, participants were men (18+ years) whose mothers carried them while living in one of two residential areas where indoor air had been contaminated by LC-PCB evaporating from building materials in subsets of the apartments. Men were considered prenatally exposed if their mother had lived in a PCB-contaminated apartment and unexposed if their mother had lived in an uncontaminated apartment for a minimum of 1 year during the 3.6 years before conception or during the first trimester. Mothers of prenatally unexposed men could not have lived in a contaminated apartment at any point. Recruitment lasted from 2017 to 2019. In total, 73 exposed and 111 unexposed men gave a blood and semen sample. Percentage differences in semen volume, sperm concentration, total sperm count, morphologically normal spermatozoa, progressively motile spermatozoa and DNA fragmentation index (DFI) between prenatally exposed and unexposed men were estimated using negative binomial regression. Associations with total and calculated free testosterone (CFT), LH and FSH were modeled using the linear regression. Odds of small testicular volume was estimated with logistic regression. Overall, the results of this study were conflicting. No differences in semen volume, sperm concentration, testosterone and CFT were observed between the groups, but there were slight indications of lower total sperm count, increased FSH and risk of small testicles, alongside lower sperm DFI and a higher proportion of normal spermatozoa in men exposed to LCB-PCBs from indoor air during fetal life. There is no apparent biologically plausible explanation for the apparently improved measures of DNA fragmentation and morphology, and these findings may have occurred purely by chance. Owing to the indirect measure of exposure, lack of adjustment for paternal factors, the potential for self-selection due to known exposure status and fertility issues, inability to take time spent away from the residence, limited statistical power and lack of comparable literature, independent replication of the study in larger cohorts is warranted. While our findings may appear reassuring for the large number of people residing and/or working in buildings with indoor air contaminated with LC-PCBs, further efforts to understand the full range of health consequences of fetal LC-PCB exposure are needed. The study was supported by the Independent Research Fund Denmark (ref no. 6110-00085B), Bispebjerg Hospital, Landsbyggefonden, Realdania (ref. no. PRJ-2017-00176), Grundejernes Investeringsfond (ref. no. 18-58) and Helsefonden (ref. no. 16-B-01-22 and 21-B-0412). K.S.H. was supported by FFIKA, Focused Research Effort on Chemicals in the Working Environment, from the Danish Government. The authors declare that they have no financial, personal or professional competing interests. Not applicable.

Does sperm DNA fragmentation affect clinical outcomes during vitrified-warmed single-blastocyst transfer cycles? A retrospective analysis of 2034 vitrified-warmed single-blastocyst transfer cycles.

Sperm DNA is essential in embryo development. The sperm DNA fragmentation index (DFI), which reflects the degree of sperm DNA fragmentation (SDF), is a crucial biomarker in evaluating the sperm quality. However, whether SDF influences the clinical outcomes after in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) remains controversial. This study aimed to investigate the relationship between sperm DNA SDF and clinical outcomes of vitrified-warmed single-blastocyst transfer cycles. A total of 2034 vitrified-warmed single-blastocyst transfer cycles (536 from ICSI and 1498 from IVF) were included in this analysis. According to the sperm DFI, all cycles were divided into two groups (DFI < 27.3% group and DFI ≥ 27.3% group). The Mann-Whitney and chi-squared tests were used to compare patient characteristics and clinical outcomes between the two groups. Furthermore, logistic regression analysis was performed to analyze the association between SDF and clinical outcomes. The chi-squared test showed no differences in positive human chorionic gonadotropin (HCG) rate, clinical pregnancy rate, miscarriage rates, and live birth rate between the two groups. Logistic regression analysis indicated that SDF was not a prognostic predictor of positive HCG, clinical pregnancy, miscarriage, and live birth. SDF was not associated with clinical outcomes either in ICSI or IVF cycles during vitrified-warmed single-blastocyst transfer cycles.

Associations of Sperm mtDNA Copy Number, DNA Fragmentation Index, and Reactive Oxygen Species With Clinical Outcomes in ART Treatments

Recent studies have suggested that sperm mitochondrial DNA copy number (mtDNA-CN), DNA fragmentation index (DFI), and reactive oxygen species (ROS) content are crucial to sperm function. However, the associations between these measurements and embryo development and pregnancy outcomes in assisted reproductive technology (ART) remain unclear. Semen samples were collected from 401 participants, and seminal quality, parameters of sperm concentration, motility, and morphology were analyzed by a computer-assisted sperm analysis system. DFI, mtDNA-CN, and ROS levels were measured using sperm chromatin structure assay, real-time quantitative polymerase chain reaction, and ROS assay, respectively. Among the participants, 126 couples underwent ART treatments, including in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI), and 79 of the couples had embryos transferred. In 401 semen samples, elevated mtDNA-CN and DFI were associated with poor seminal quality. In 126 ART couples, only mtDNA-CN was negatively correlated with the fertilization rate, but this correlation was not significant after adjusting for male age, female age, seminal quality, ART strategy, number of retrieved oocytes, controlled stimulation protocols, and cycle rank. Regarding pregnancy outcomes, sperm mtDNA-CN, ROS, and DFI were not associated with the clinical pregnancy rate or live birth rate in 79 transferred cases. In conclusion, increased mtDNA-CN and DFI in sperm jointly contributed to poor seminal quality, but sperm mtDNA-CN, ROS, and DFI were not associated with clinical outcomes in ART.

Exposure to polycyclic aromatic hydrocarbons and nicotine, and associations with sperm DNA fragmentation.

BackgroundTobacco smoking has been reported to cause DNA fragmentation and has been suggested to cause mutations in spermatozoa. These effects have been ascribed to the action of polycyclic aromatic hydrocarbons (PAH) present in the smoke. Simultaneously, DNA fragmentation has been associated with mutagenesis.ObjectiveThe aim of this study was to investigate whether levels of urinary biomarkers of PAH and nicotine exposure were associated with sperm DNA fragmentation.MethodsIn the urine of 381 men recruited from two cohorts of young men (17–21 years old) from the general Swedish population, the PAH metabolites 1‐hydroxypyrene and 2‐hydroxyphenanthrene, as well as the nicotine metabolite cotinine, were measured. The sperm DNA fragmentation index (DFI) was analysed using the sperm chromatin structure assay. Associations between the DFI, and PAH metabolite levels as continuous variables as well as in quartiles, were studied by general linear models adjusted for abstinence time. A similar analysis was carried out for cotinine levels, according to which the men were categorised as “non‐smoking” (n = 216) and “smoking” (n = 165).ResultsNo association was found between levels of any of the three biomarkers and DFI, either as a continuous variable (p = 0.87–0.99), or when comparing the lowest and the highest quartiles (p = 0.11–0.61). The same was true for comparison of men categorised as non‐smoking or smoking (DFI 11.1% vs. 11.8%, p = 0.31).DiscussionWe found no evidence of PAH or nicotine exposure to be associated with DFI, which does not exclude that these exposures may have other effects on sperm DNA.ConclusionIn these young men, levels of biomarkers of nicotine and PAH exposure were not associated with DFI.