Sperm DNA Fragmentation Testing Research Articles

Evaluation of sperm DNA fragmentation index among infertile individuals: a comprehensive analysis of the associated factors—a cross-sectional study

BackgroundEnsuring the integrity of sperm DNA is of paramount importance for the production of healthy offspring. The process of sperm formation involves intricate molecular adjustments that condense and safeguard its genetic material. However, the risk posed by sperm DNA fragmentation (SDF) can impede natural and assisted fertility and may arise from various testicular and post-testicular factors. While there are recommendations against routine SDF testing, recent research has sparked debate by demonstrating the positive impact of such testing on fertility outcomes.ObjectiveTo identify the factors associated with SDF among infertile individuals.Methods and materialsThe medical records of infertile men referred to our hospital were comprehensively reviewed and included in the study as they met the inclusion criteria. Patients dissatisfied with participation in the study or those with incomplete medical records were excluded. The association of SDF, sperm analysis, and demographic characteristics were evaluated for further investigation.ResultsThe average age of participants was 36.1 ± 8.1 years. The average semen volume was 2.5 ± 0.3 cc with a semen pH of 7.4 ± 0.2. The average sperm count was 10.9 ± 0.7 million, sperm motility was 36.9 ± 2.4%, and normal sperm morphology was 41.7 ± 2.7%. SDF levels below 15% were seen in 27.9%, between 15 and 30% in 32.4%, and above 30% in 39.7% of patients. Significant associations were found between the SDF and various factors in sperm analysis, including sperm count (P < 0.001), semen volume (P < 0.001), semen pH (P < 0.001), sperm motility (P < 0.001), normal sperm morphology (P < 0.001), and age (P < 0.001). Moreover, alcohol consumption (P = 0.04), smoking (P = 0.01), and the presence of varicocele (P = 0.03) were significantly associated with.ConclusionThis study emphasizes the significance of SDF among individuals experiencing infertility and investigates its correlation with various sperm test parameters. It also sheds light on the vital roles played by factors such as age, smoking, alcohol consumption, and the presence of varicocele, which are highly likely to cause variations in SDF levels. The results of our research will add to the existing pool of knowledge on infertility and potentially have implications for clinical practice and the care of patients.

One out of two idiopathic infertile men has pathologic sperm DNA fragmentation values: Potential implications for clinical practice.

To investigate the distribution of sperm DNA fragmentation (SDF) values and their association with clinical and seminal parameters in idiopathic infertile men. Data from 3224 primary infertile men (belonging to couples having failed to conceive a pregnancy within 12 months) who underwent a thorough diagnostic work-up were analysed. A SDF value ≥ 30% (according to Sperm Chromatin Structure Assay) was considered pathologic. We excluded: (1) men with genetic abnormalities; (2) men with history of cryptorchidism; (3) men with biochemical hypogonadism; (4) men with clinical varicocele; and (5) men with other possible known aetiological factors. Descriptive statistics and logistic regression analyses were used to describe the whole cohort. Of all, 792 (23%) men with at least one abnormal WHO semen parameter but without any identified aetiologic factor for infertility, were considered as idiopathic infertile men. Of 792, 418 (52.7%) men had SDF≥30%. Men with pathologic SDF were older (p = .02), had higher Follicle-stimulating hormone(FSH) (p = .04) but lower total testosterone (p = .03) values than those with SDF<30%. The homoeostatic model assessment index for insulin resistance (HOMA-IR) was higher in men with SDF≥30% (p = .01). Idiopathic infertile men with SDF≥30% presented with lower sperm concentration (p < .001) and lower progressive sperm motility (p < .01) than those with SDF < 30%. Logistic regression analysis revealed that older age (OR: 1.1, p = .02) and higher HOMA-IR score (OR: 1.8, p = .03) were associated with SDF ≥ 30%, after accounting for FSH and sperm concentration values. Approximately half of infertile men categorized as idiopathic had pathologic SDF values. Idiopathic infertile men with pathologic SDF showed worse clinical, hormonaland semen parameters than those with normal SDF values. These results suggest that including SDF testing could be clinically relevant over the real-life management work-up of infertile men.

Sperm DNA fragmentation and infertility: a narrative review.

The purpose of this narrative review is to provide a practical understanding of sperm DNA fragmentation (SDF) in the management of male infertility. A search for systematic reviews and meta-analyses (SRMA) on SDF between April 1st, 2018 and April 1st, 2023 was performed using PubMed and articles were selected as per their relevance to the topic. Guidelines from major societies were also reviewed. Three clinical cases are reported and discussed. The search initially identified 80 articles. We selected 13 SRMAs based on their relevance to the topic. Of the 13 SRMAs, 7 evaluated the effect of SDF on assisted reproductive technology (ART) outcomes and recurrent pregnancy loss, 3 studied the effect of varicocele repair on SDF, and 3 evaluated the role of SDF involving lifestyle and environmental health factors including body mass index and male factor treatment strategies. Evidence suggests that increased SDF has a negative impact on natural pregnancy and ART outcomes. SDF testing may be particularly important in the infertility evaluation of men with varicoceles, idiopathic or unexplained infertility, recurrent pregnancy loss, or previous ART failure. Further studies are needed on SDF testing and the implications it can have on male factor infertility and pregnancy outcomes as well as its implementation in the setting of ART.

P-717 Add-ons availability and quality of online information provided by Brazilian IVF clinics

Abstract Study question How do Brazilian fertility clinic websites advertise four IVF add-ons: assistedhatching (AH), preimplantation genetic testing for aneuploidies (PGT-A), ERA and sperm DNA fragmentation test. Summary answer Available information, about ivf add-ons in Brazil, on researched websites is heterogeneous and is not evidence-based according to the last published recommendations. What is known already Over 1 billion people worldwide use online resources to obtain health-related information. People struggling with infertility are increasingly turning to online resources for infertility-related content and social support. “Add-on” procedures are actively promoted on some fertility clinic websites as means to improve IVF success rates. However, the actual contribution of these interventions to IVF success rates remains debatable. ESHRE has recently issued 42 recommendations were formulated on the use of add-ons in IVF. At present, no study has evaluated the type and quality of the information provided on the Brazilian IVF clinics’ websites regarding the merits of “add-ons. Study design, size, duration Prospective evaluation of the availability and quality of information on four common add-ons provided by IVF clinics in the southeast of Brazil between October-November 2023. Clinics were identified using the national registry (SisEmbrio) as well as through manual online search. As only public online available information was used no ethical approval was necessary. Statistical analyses were restricted to descriptive of the frequency of each add-on and frequency of claims made in relation to their use. Participants/materials, setting, methods One reviewer took screenshots of the included clinic pages and claims and a second reviewer double-checked everything. The folowing information was colletced: availability of of the four add-ons; information on the effectiveness on: implanation rate(IR), pregnancy rate (PR) or live-birth rate (LBR); information on the scientifc uncertainty of the use of add-on and possible negative impact; for PGT-A if the clinic informed that the data obtained should be confirmed by prenatal diagnostic testing. Main results and the role of chance The websites of 122 IVF clinics were included. Nine clinics had no website or were offline. PGT-A was the most frequent available add-on (n = 77; 68%) but only two of them (2,5%) informed on PR. There was no information on IR nor LBR. Thirteen clinics (16,8%) acknowledged it could have adeverse effects but only one mentioned the scientifc uncertainty related to its use. Forty (35%) websites displayed AH with 7 reporting on possible negative effects but none disclosed success rates (PR, IR or LBR). Eight clinics admitted AH had uncertain benefits and seven noted possible negative impact. The sperm DNA fragmentation test was presented in 37 (33%) websites but none reported on potential adverse effects nor success rates in terms of PR, IR nor LBR. No website provided information on the uncertainty of the benefit of the test. The ERA-test was advertised in 28 (25%) websites with no information on its benefits on sucess rates. Three clinics advised it could have a negative impact and six agreed its effectiveness was unclear. The cost was never pesented for any of the add-ons researched in accordance to national regulations. Of note, 36 websites advertised other add-ond such as MSOME, ZYMOT and endometrial scrathcing. Limitations, reasons for caution The study was restricted to 122 clinics websites in the southeast of Brazil. Evalaution was restricted to four commons add-ons and the quality of the information provided could not be assessed as the clinics follow no specific regulation on how to advertise scientifc information to their patiens and general public. Wider implications of the findings The information provided by Brazilian IVF clinics could be misleading. The use of add-ons is surrounded by scientific uncertainly related to their benefits and cost-effectiveness therfeore it is of utmost importance that clinics reviewt heir websites and offer sound scientific data to patients and the general public as well. Trial registration number Not applicable

P-016 Prevalence of high oxidative stress and poor sperm DNA fragmentation levels in different groups of patients

Abstract Study question What is the prevalence of patients with high oxidation-reduction potential (ORP) and high sperm DNA fragmentation (SDF) in different patient groups with and without varicocele? Summary answer A large percentage of normozoospermic patients and patients who are normally considered of manageable in assisted reproduction show high SDF and/or high ORP. What is known already Standard semen analysis, even according to WHO criteria, is very limited in predicting male fertility potential. Oxidative stress is widely recognized as a major contributor to male infertility. Therefore, sperm DNA fragmentation (SDF) is increasingly recommended as additional test in diagnostics. High levels of oxidative stress and SDF have also been reported in patients with varicocele. Yet, SDF and oxidative stress tests are not routinely tested in ART laboratories. Apart from the cost and/or complexity of these tests, a reason for the lack of usage is the lack of knowledge of the prevalence. Study design, size, duration This is a retrospective analysis of data obtained from 2,946 patients attending the Department of Urology, Hamad Medical Corporation, Doha, Qatar, for fertility diagnosis from January to December 2018. For this analysis only patients (n = 773) who had a full data set for sperm DNA fragmentation (SDF) and seminal oxidative stress as measured by means of the oxidation-reduction potential (ORP) were included. Participants/materials, setting, methods Apart from SDF and ORP, standard semen parameters (ejaculate volume, sperm concentration, total and progressive motility, and normal sperm morphology), and the presence of varicocele were determined. Semen parameters were determined according to WHO 2010 criteria. SDF was tested using the Halosperm G2 test (Halotech DNA SL, Madrid, Spain), while ORP was determined using the MiOXSYS® (Aytu BioScience, Englewood, CO). Patients were categorized as normo-, oligo-, astheno-, and teratozoospermic. Main results and the role of chance The mean sperm concentration of the patients was 33.5 × 106 sperm/mL with a mean total motility of 50.3%. The overall percentage of patients with high (&amp;gt;20% SDF) SDF was 57.4% and those with high (≥1.34 mV/106 sperm/mL) ORP 63.4%. Varicocele was diagnosed in 14.0% of the patients. In normozoospermic patients, 26.8% had high SDF, whereas in the non-normozoospermic group this was 59.8% (P = 0.0108). In non-oligozoospermic (53.7%), non-asthenozoospermic (34.7%) and non-teratozoospermic (43.4%) groups, SDF was significantly higher. High ORP values were recorded in 35.7% of normozoospermic, 53.4% non-oligozoospermic, 47.2% of non-asthenozoospermic, and 49.7% of teratozoospermic patients. The difference between the prevalence of high ORP in the normozoospermic (35.7%) and the non-normozoospermic group is significant (P = 0.0063). When both tests (high SDF or high ORP) were considered, 50.0% of normozoospermic, 76.9% of non-oligozoospermic, 59.7% of non-asthenozoospermic, and 71.2% of non-teratozoospermic patients showed seminal pathology. High SDF was detected in 59.3% of varicocele patients, while 50.0% of varicocele patients had high ORP values. In patients without varicocele, 57.1%, 65.6%, and 82.7% had high SDF, high ORP, and high SDF or high ORP, respectively. These percentages were significantly (P &amp;lt; 0.0003) higher than for the corresponding normal groups. Limitations, reasons for caution Although the overall sample size of 773 is reasonably high, after sub-grouping the sample size is much smaller. Since this study only analyzed the SDF and ORP in patients attending and andrological outpatient clinic, the results could be different if patients in IVF and/or ICSI treatment cycles were analyzed. Wider implications of the findings This study highlights that complementation of semen analysis with SDF and oxidative stress testing will identify a large percentage of patients who would benefit from andrological treatment before assisted reproductive techniques (ART). This approach would result in better chances of these patients for successful ART. Trial registration number not applicable

Contribution of sperm DNA fragmentation testing by flow cytometry in medically assisted reproduction procedures

Contribution of sperm DNA fragmentation testing by flow cytometry in medically assisted reproduction procedures

SPERMATIC DNA FRAGMENTATION INDEX OF INFERTILE MEN: INTEREST IN THE CHOICE OF PMA TECHNIQUE PRELIMINARY STUDY

Introduction: Infertility constitutes a real public health problem. In men, the examinations involved in the choice of the PMA technique are sometimes insufficient in relation to the real procreation potential of the spermatozoa which could be detected by the sperm DNA fragmentation test. This preliminary study aimed to highlight the benefit of taking into account the sperm DNA fragmentation test in the choice of the proposed PMA technique. Material and Methods: This was a preliminary descriptive cross-sectional study carried out in a private spermiology laboratory in Abidjan. It included 52 patients who came for consultation for infertility and who gave their written consent to participate in the study. Spermogram, spermocytogram, migration survival (TMS) test and DNA fragmentation index were studied. The DNA fragmentation index (DFI) was determined by the Sperm Chromatin Dispersion (SCD) technique. Results: The majority of patients were aged between 35 and 45 years (47%) with an average age of 45 years (+/-8). The most common spermogram abnormalities were mobility abnormalities. 52% of patients had a high fragmentation index. A high fragmentation index was found in a patient with normal sperm standard parameters. Likewise, 8 patients for whom intrauterine insemination had been recommended according to the TMS result had a fragmentation index greater than 30%. Conclusion: The first-line request for the fragmentation index in the exploration of male infertility must be studied since it could take into account the integrity of the genetic material, a limiting factor in the current tests used for the choice of the PMA technique.

Male Infertility: New Developments, Current Challenges, and Future Directions.

There have been many significant scientific advances in the diagnostics and treatment modalities in the field of male infertility in recent decades. Examples of these include assisted reproductive technologies, sperm selection techniques for intracytoplasmic sperm injection, surgical procedures for sperm retrieval, and novel tests of sperm function. However, there is certainly a need for new developments in this field. In this review, we discuss advances in the management of male infertility, such as seminal oxidative stress testing, sperm DNA fragmentation testing, genetic and epigenetic tests, genetic manipulations, artificial intelligence, personalized medicine, and telemedicine. The role of the reproductive urologist will continue to expand in future years to address different topzics related to diverse questions and controversies of pathophysiology, diagnosis, and therapy of male infertility, training researchers and physicians in medical and scientific research in reproductive urology/andrology, and further development of andrology as an independent specialty.

Changing Trends of Tubercular Assault on Reproduction

Background and Objectives. Genital tuberculosis (GTB) is a rare disease in developed countries but is an important cause of subclinical chronic pelvic inflammatory state and infertility in 1–18% of women, being as high as 18 % in India. What we are observing nowadays GTB is changing its way of assault to reproductive process less by infection but more by infestation of the genital tract and thereby leading to harmful molecular changes or may be immunomodulation locally. Reduction of infection may be due to increased surveillance, treatment or use of antibiotics for other reasons indiscriminately, which may make the bacteria dormant. It is being observed as a silent invader of the genital tract of both the female and male partners. It is to be kept in mind that mycobacteria, non- pathogenic to humans, may also cause molecular assault on reproduction. At present, our cohort of patients can be categorized in three groups a) Frank tuberculosis b) LGTB infection c) LGTB infestation There has been extensive research about the involvement of genital tuberculosis in female infertility. Various studies have been undertaken to screen and detect latent genital tuberculosis in apparently normal infertile couples intervening fertility at molecular level. The adverse effects of latent genital tuberculosis, as found in current researches, on the female and male reproductive process has been addressed in the current review. Methods Endometrial aspirates at mid-luteal phase and samples of menstrual blood were collected aseptically from each of those women enrolled in this study and stored at - 80°C for DNA-PCR (Polymerase chain reaction test) and cytokine study. Multiplex polymerase chain reaction (PCR) were performed. DNA-PCR test and sperm DNA fragmentation tests were carried out in semen samples, collected according to WHO guidelines, from male partners in question. Observation. It is being observed that pro-inflammatory cytokines showed significantly different values in TB-PCR- positive and TB-PCR-negative groups. Sperm DNA fragmentation index also appears to be quite high in TB- PCR positive males.Conclusion. LGTB infestation has been found to be responsible for unfavourable molecular changes in reproductive tract and thereby causing a negative impact on fertility. Keywords: latent genital tuberculosis, infertility, cytokine, tubercular infestation, tuberculosis- polymerase chain reaction,

Effects of Varicocele Repair on Sperm DNA Fragmentation and Seminal Malondialdehyde Levels in Infertile Men with Clinical Varicocele: A Systematic Review and Meta-Analysis.

Varicoceles can be a source of elevated seminal oxidative stress (OS) and sperm DNA fragmentation (SDF). However, it remains unclear whether varicocele repair (VR) could reduce these parameters. This systematic review and meta-analysis (SRMA) aims to investigate the impact of VR on SDF and seminal malondialdehyde (MDA). A literature search was performed in Scopus, PubMed, Ovid, Embase, and Cochrane databases. This SRMA included randomized controlled trials and observational studies reporting the pre- and postoperative levels of SDF and seminal OS in infertile men with clinical varicocele that underwent VR. Subgroup analyses included techniques of VR and SDF testing. The effect size was expressed as standardized mean difference (SMD). Out of 1,632 abstracts assessed for eligibility, 29 studies with 1,491 infertile men were included. The analysis showed a significant reduction in SDF after VR, compared to preoperative values (SMD -1.125, 95% confidence interval [CI] -1.410, -0.840; p<0.0001) with high inter-study heterogeneity (I²=90.965%). Reduction in SDF was evident with microsurgical technique and non-microsurgical inguinal approaches (SMD -1.014, 95% CI -1.263, -0.765; p<0.0001, and SMD -1.495, 95% CI -2.116, -0.873; p<0.0001), respectively. Reduction in SDF was significant irrespective of testing was done by sperm chromatin dispersion (SMD -2.197, 95% CI -3.187, -1.207; p<0.0001), sperm chromatin structure assay (SMD -0.857, 95% CI -1.156, -0.559; p<0.0001) or TUNEL (SMD -1.599, 95% CI -2.478, -0.719; p<0.0001). A significant decrease in seminal MDA levels was observed following VR (SMD -2.450, 95% CI -3.903 to -0.997, p=0.001) with high inter-study heterogeneity (I²=93.7%). Using pre- and post-intervention data, this SRMA indicates a significant reduction in SDF and seminal MDA levels in infertile men with clinical varicocele treated with VR. These findings may have important implications for the future management of this selected group of infertile patients.

The Importance of Discordant Follicle Stimulating Hormone and Inhibin B Levels in Primary Infertile Men: Findings from a Cross-Sectional Study.

We aimed to investigate the relationship between follicle stimulating hormone (FSH) and inhibin B (InhB). Data from 1,230 consecutive men presenting for primary couple's infertility were analyzed. Health-significant comorbidities were scored with Charlson comorbidity index. Quartiles of FSH and InhB were considered to determine threshold values. Descriptive statistics and logistic regression models tested association between FSH and InhB values. Overall, 1,080 (87.8%) men had concordant FSH and InhB values. Conversely, 150 patients (12.2%) had discrepancies in FSH and InhB, with 78 (6.3%) and 72 (5.9%) men reporting both low and high FSH and InhB values, respectively. Infertile men with discordant values were younger (median [interquartile range] 38.0 years [34-41 years] vs. 36.0 years [31-40 years]); had smaller testicular volume (TV) (12 mL [10-15 mL] vs. 15 mL [12-20 mL]); and, had more frequently a sperm DNA fragmentation test >30% (179 [59.1%] vs. 40 [78.4%]) than those with concordant values (all p<0.05). Moreover, a higher frequency of previous cryptorchidism (27.3% vs. 11.9%), lower sperm concentration (3.0 million/mL [0.9-11.0 million/mL] vs. 13.8 million/mL [3.1-36.0 million/mL]), lower progressive sperm motility rates (12.0% [5.0%-25.3%] vs. 20.0% [7.0%-36.0%]), and greater rates of non-obstructive azoospermia (36.4% vs. 23.9%) were found in men with discordant FSH and InhB values (all p≤0.005). At multivariable logistic regression analysis, higher body mass index (odds ratio [OR], 1.08; p=0.001), smaller TV (OR, 0.91; p<0.001), and a history of cryptorchidism (OR, 2.49; p<0.001) were associated with discordant FSH and InhB values. More than one out of ten infertile men had discordant FSH and InhB values in the real-life setting showing worse clinical profiles than those with concordant levels. Smaller TV and history of cryptorchidism could be used as clinical markers to better tailor the need to test InhB.

#108 : Methods of Sperm DNA Fragmentation (SDF) Testing and Interpretation: Results from the Global Andrology Forum Survey

Background and Aims: Sperm DNA fragmentation (SDF) is caused by apoptosis, oxidative stress or incomplete chromatin maturation and can result in male infertility and poor reproductive outcomes. Despite growing trends, there are no specific recommendations on specific diagnostic tests used, assay cut-offs or abstinence periods prior to SDF testing. Therefore, we conducted a global survey to evaluate the approaches used to diagnose SDF, towards a consensus in the field of SDF research and clinical management. Method: The survey collected cross-sectional global data from 55 countries following the CHERRIES protocol. 436 respondents included clinicians, researchers and reproductive biologists from academic, public and private health sectors, all of whom utilize SDF testing in their routine patient care and practice (Fig. 1). Results: Survey results showed that TUNEL (28.6%), SCSA (24.1%), SCD (19.1%) and Comet (5.3%) were most frequently used to diagnose SD (Fig. 2A). Assays were chosen primarily due to their availability (70.0%), cost (44.05%) and accuracy (35.48%). Published evidence on assays (25.71%), training of staff (25.48%), cost to patients (21.67%), assay time (18.81%), ease of interpretation (18.33%) and performance (17.38%) were taken lesser into consideration when selecting SDF diagnostic tests (Fig. 2B). Assay cut-offs of “30% or higher” (33.7%) were most frequently used to diagnose SDF (Fig. 2C). Finally, an abstinence period of 3-5 days was mostly recommended (53.6%; Fig. 2D). The results of this survey provide valuable data on SDF assays used in clinical practice as well as factors affecting assay choices such as availability of resources, personnel and complexity in assay processing. As decreased abstinence period is associated with lower SDF, testing is recommended with an abstinence period of less than five days. We observed that there was a lack of standard cut-off values, and every laboratory should establish its own reference values based on predictive values for fertility outcomes. Conclusion: We present new data associated with SDF testing in real-world practice, on a global scale. The trends and conclusions derived from the study are recommended for inclusion into the 6[Formula: see text] Edition WHO Manual, towards optimized patient care.

Unraveling the Impact of Sperm DNA Fragmentation on Reproductive Outcomes.

In recent years, there has been a growing interest in identifying subcellular causes of male infertility, and sperm DNA fragmentation (SDF) research has been at the forefront of this focus. DNA damage can occur during spermatogenesis due to faulty chromatin compaction or excessive abortive apoptosis. It can also happen as sperm transit through the genital tract, often induced by oxidative stress. There are several methods for SDF testing, with the sperm chromatin structure assay, terminal deoxynucleotidyl transferase d-UTI nick end labeling (TUNEL) assay, comet assay, and sperm chromatin dispersion test being the most commonly used. Numerous studies strongly support the negative impact of SDF on male fertility potential. DNA damage has been linked to various morphological and functional sperm abnormalities, ultimately affecting natural conception and assisted reproductive technology outcomes. This evidence-based review aims to explore how SDF influences male reproduction and provide insights into available therapeutic options to minimize its detrimental impact.

P-059 Sperm DNA fragmentation (SDF) after cryopreservation and sperm selection: implications for clinical pregnancies and live births after intrauterine insemination with donor sperm

Abstract Study question Can SDF assessment pre-cryopreservation, post-thaw and after post-thaw sperm selection predict clinical outcome in a heterologous intra-uterine donor insemination program (IUI)? Summary answer SDF increased post-cryopreservation in donor sperm and after post-thaw density gradient without affecting clinical pregnancy, live birth and miscarriage rates. What is known already Sperm cryopreservation is effective for preservation of male fertility and facilitates the storage of donor semen, while infectious disease screening can be completed and confirmed negative. Sperm viability and motility are the most vulnerable parameters during the freeze-thaw process. Inherent reactive oxygen species may induce SDF via oxidative stress. Controversial results are reported regarding induction of SDF after cryopreservation due to differences in cryo tolerance, methods of cryopreservation and methods used to evaluate SDF. Sperm selection post-thaw is indispensable to select a normal motile fraction for insemination. Again, results are controversial regarding influence on SDF levels after sperm selection post-thaw. Study design, size, duration This was a prospective, observational study. Between March 2015 and March 2019, eighteen potential sperm donors were screened via a three step plan – semen assessment, medical assessment and serological blood tests. In accordance with the Belgian legislation (2007), a sperm donor was matched to obtain a maximum of 6 pregnancies in acceptor women/couples, allowing for more than one offspring/woman or couple. A total of 106 acceptors were matched for heterologous intra-uterine insemination. Participants/materials, setting, methods Semen samples were collected after 2-7 days abstinence and standard semen parameters analyzed within 60 mins after ejaculation. Within the same time frame SDF test was performed using TUNEL assay both before and after cryopreservation and after a two-step discontinuous density gradient centrifugation post-thaw. Samples were cryopreserved by the conventional slow freezing method using sperm freeze solution. A yield of ≥ 2 M progressive spermatozoa post-thaw after selection was sufficient to contemplate IUI. Main results and the role of chance Mean female age at first cycle was 33.9 ± 4.1 years and donor age 28.5 ± 5.6 years. Out of 429 cycles, 100 (23.3%) resulted in clinical pregnancy. Excluding three ongoing pregnancies, we counted 75 live births (17.6% of cycles or 77.3% of pregnancies), while 21 pregnancies ended in miscarriage (4.9% of cycles or 21.6% of pregnancies) and 1 resulted in stillbirth. Progressive motility (59.3 ± 12.5%) decreased significantly after cryopreservation (30.0 ± 13.2%; p &amp;lt; 0.001), but increased post-thaw after density gradient centrifugation (61.6 ± 16.0%; p &amp;lt; 0.001). On the contrary, SDF (12.0 ± 5.9%) increased post-cryopreservation (26.3 ± 14.5%; p &amp;lt; 0.001) and further increased after sperm selection (34.9 ± 22.1%; p = 0.04). Using multilevel mixed-effects logistic regression (Odds ratio [95%CI]), we found that female age significantly influenced clinical pregnancy (0.911 [0.847-0.981]; p = 0.01), live birth (0.894 [0.834-0.959]; p = 0.002) and miscarriage rates (1.180 [1.033-1.347]; p = 0.015). There was no significant age-adjusted effect of SDF post-thaw after density gradient selection on clinical pregnancy (1.007 [0.994-1.021]; p = 0.30), live birth (1.001 [0.988-1.014]; p = 0.84) or miscarriage (1.021 [0.997-1.046]; p = 0.08). Median time-to-live birth was 4 cycles. Analyses of time-to-live birth with mixed-effects Cox models revealed no significant association with progressive motility; SDF before or after cryopreservation and after sperm selection post-thaw. Limitations, reasons for caution Results concern a small donor population, with high normal semen parameters and should therefore be extrapolated cautiously to subfertile men with normal and subnormal semen parameters. Wider implications of the findings In a donor IUI program with strict selection on conventional sperm parameters, differences in SDF before and after cryopreservation and gradient centrifugation do not change IUI outcomes, although cryopreservation and centrifugation increase SDF. Findings could be useful for the optimization of sperm freezing and selection in different assisted reproductive procedures. Trial registration number not applicable

Performing a sperm DNA fragmentation test in addition to semen examination based on the WHO criteria can be a more accurate diagnosis of IVF outcomes

BackgroundWe analyzed the sperm DNA fragmentation index (DFI) and general semen test based on the World Health Organization (WHO) criteria and compared the two tests using semen factors. In addition, we examined whether DFI is a reliable parameter associated with in vitro fertilization (IVF) outcomes.MethodsSperm chromatin dispersion (SCD) and general semen tests were conducted in accordance with the WHO 2010 guidelines, and correlations between the two tests were investigated. The WHO criteria were set as the cutoff values for each of the following factors: semen volume, concentration, total sperm count, motility, and normal morphology, and compared with the DFI results.ResultsThe subjects had a mean sperm DFI of 15.3% ± 12.6%, and the DFI increased with age. In contrast, motility and normal morphology decreased as the DFI increased. Patients who satisfied the WHO criteria in terms of concentration, total sperm count, and motility had a significantly lower DFI than those who did not satisfy the criteria. Therefore, evaluation with a general semen test based on the WHO criteria should be regarded as a qualitative evaluation of all factors other than semen volume and normal morphology.ConclusionsHigh DFI (≥ 30%) caused a low blastocyst development rate following intracytoplasmic sperm injection. Male infertility due to DFI should be suspected when IVF results are poor despite normal semen findings based on the WHO criteria. The results of this study suggest that the SCD test may more accurately evaluate the correlation between IVF clinical outcomes and male infertility. Therefore, it is important to focus on DFI measurements.

Novel sperm chromatin dispersion test with artificial intelligence-aided halo evaluation: A comparison study with existing modalities.

Sperm chromatin dispersion test is a common and inexpensive technique to assess sperm DNA fragmentation, but its subjectivity in assessing a small number of spermatozoa is a disadvantage. To study the efficacy of a new sperm chromatin dispersion test kit (R10) combined with an artificial intelligence-aided halo-evaluation platform (X12) and compare the results to those of existing sperm DNA fragmentation testing methods. Semen samples from normozoospermic donors (n=10) and infertile men with abnormal semen parameters (n=10) were enrolled. DNA fragmentation indices were examined by multiple assays, including R10, Halosperm G2 (G2), sperm chromatin structure assay, and terminal deoxynucleotidyl transferase deoxynucleotidyl transferase nick end labeling. In R10 assay, the DNA fragmentation indices were obtained both manually (manual R10) and by X12 (AI-R10). The obtained DNA fragmentation indices were analyzed by agreement analyses. The DNA fragmentation indices obtained by manual R10 and those obtained by AI-R10 showed a strong significant correlation (r=0.97, p<0.001) and agreement. The number of spermatozoa evaluated by AI-R10 was 2078 (680-5831). The DNA fragmentation indices obtained by manual R10 and AI-R10 both correlated with those of G2 (r=0.90, p<0.001; r=0.88, p<0.001). Between the AI-R10 and G2 results, Passing-Bablok regression showed no systematic or proportional difference, and Bland-Altman plots revealed overall agreement and a mean bias of 6.3% with an SD of 6.9% (95% limit of agreement: -7.2% to 19.9%). AI-R10 and sperm chromatin structure assays showed systematic differences with a mean bias of -1.9%, while AI-R10 and terminal deoxynucleotidyl transferase deoxynucleotidyl transferase nick end labeling revealed proportional differences with a mean bias of -10.7%. The novel sperm chromatin dispersion kit and artificial intelligence-aided platform demonstrated significant correlation and agreement with existing sperm chromatin dispersion methods by assessing greater number of spermatozoa. This technique has the potential to provide a rapid and accurate assessment of sperm DNA fragmentation without technical expertise or flow cytometry.

Controversy and Consensus on Indications for Sperm DNA Fragmentation Testing in Male Infertility: A Global Survey, Current Guidelines, and Expert Recommendations.

Sperm DNA fragmentation (SDF) testing was recently added to the sixth edition of the World Health Organization laboratory manual for the examination and processing of human semen. Many conditions and risk factors have been associated with elevated SDF; therefore, it is important to identify the population of infertile men who might benefit from this test. The purpose of this study was to investigate global practices related to indications for SDF testing, compare the relevant professional society guideline recommendations, and provide expert recommendations. Clinicians managing male infertility were invited to take part in a global online survey on SDF clinical practices. This was conducted following the CHERRIES checklist criteria. The responses were compared to professional society guideline recommendations related to SDF and the appropriate available evidence. Expert recommendations on indications for SDF testing were then formulated, and the Delphi method was used to reach consensus. The survey was completed by 436 experts from 55 countries. Almost 75% of respondents test for SDF in all or some men with unexplained or idiopathic infertility, 39% order it routinely in the work-up of recurrent pregnancy loss (RPL), and 62.2% investigate SDF in smokers. While 47% of reproductive urologists test SDF to support the decision for varicocele repair surgery when conventional semen parameters are normal, significantly fewer general urologists (23%; p=0.008) do the same. Nearly 70% would assess SDF before assisted reproductive technologies (ART), either always or for certain conditions. Recurrent ART failure is a common indication for SDF testing. Very few society recommendations were found regarding SDF testing. This article presents the largest global survey on the indications for SDF testing in infertile men, and demonstrates diverse practices. Furthermore, it highlights the paucity of professional society guideline recommendations. Expert recommendations are proposed to help guide clinicians.

Technical Aspects and Clinical Limitations of Sperm DNA Fragmentation Testing in Male Infertility: A Global Survey, Current Guidelines, and Expert Recommendations.

Sperm DNA fragmentation (SDF) is a functional sperm abnormality that can impact reproductive potential, for which four assays have been described in the recently published sixth edition of the WHO laboratory manual for the examination and processing of human semen. The purpose of this study was to examine the global practices related to the use of SDF assays and investigate the barriers and limitations that clinicians face in incorporating these tests into their practice. Clinicians managing male infertility were invited to complete an online survey on practices related to SDF diagnostic and treatment approaches. Their responses related to the technical aspects of SDF testing, current professional society guidelines, and the literature were used to generate expert recommendations via the Delphi method. Finally, challenges related to SDF that the clinicians encounter in their daily practice were captured. The survey was completed by 436 reproductive clinicians. Overall, terminal deoxynucleotidyl transferase deoxyuridine triphosphate Nick-End Labeling (TUNEL) is the most commonly used assay chosen by 28.6%, followed by the sperm chromatin structure assay (24.1%), and the sperm chromatin dispersion (19.1%). The choice of the assay was largely influenced by availability (70% of respondents). A threshold of 30% was the most selected cut-off value for elevated SDF by 33.7% of clinicians. Of respondents, 53.6% recommend SDF testing after 3 to 5 days of abstinence. Although 75.3% believe SDF testing can provide an explanation for many unknown causes of infertility, the main limiting factors selected by respondents are a lack of professional society guideline recommendations (62.7%) and an absence of globally accepted references for SDF interpretation (50.3%). This study represents the largest global survey on the technical aspects of SDF testing as well as the barriers encountered by clinicians. Unified global recommendations regarding clinician implementation and standard laboratory interpretation of SDF testing are crucial.

Sperm DNA fragmentation testing in clinical management of reproductive medicine.

Approximately 8%-12% of couples worldwide face infertility, with infertility of individuals assigned male at birth (AMAB) contributing to at least 50% of cases. Conventional semen analysis commonly used to detect sperm abnormalities is insufficient, as 30% of AMAB patients experiencing infertility show normal results in this test. From a genetic perspective, the assessment of sperm DNA fragmentation (SDF) is important as a parameter of sperm quality. In this narrative study, we review and discuss pathophysiological causes, DNA repair mechanisms, and management of high SDF. We then summarize literature exploring the association between SDF and reproductive outcomes. Recent systematic reviews and meta-analyses have revealed a significant association between high SDF in AMAB individuals and adverse reproductive outcomes including embryo development, natural conception, intrauterine insemination, and invitro fertilization. However, the association with live birth rates and pregnancy rates following intracytoplasmic injection remains inconclusive. The disparities among quantitative assays, inconsistent reference range values, absent high-quality prospective clinical trials, and clinical heterogeneity in AMAB patients with elevated SDF represent the main limitations affecting SDF testing. The evaluation and management of SDF plays an important role in a subset of AMAB infertility, but widespread integration into clinical guidelines will require future high-quality clinical trials and assay standardization.

Is There a Relationship between Sperm DNA Fragmentation and Intra-Uterine Insemination Outcome in Couples with Unexplained or Mild Male Infertility? Results from the ID-Trial.

Sperm DNA fragmentation has been proposed as a candidate test for the assessment of sperm function on the premise that damage to the sperm chromatin is associated with a detrimental reproductive outcome. The objective of our study was to investigate whether sperm DNA fragmentation testing has a prognostic value, and thus can play a pivotal role in selecting future patients for intra-uterine insemination (IUI) therapy. This was a prospective cohort study conducted in a University Hospital setting. SDF was measured through TUNEL assay on the fresh semen sample presented at diagnosis and at insemination in couples with idiopathic/mild male infertility undergoing natural cycle IUI treatment. The generalized estimating equation (GEE)-model and multivariable model were used to analyze the probability of live birth and clinical pregnancy, respectively. ROC analysis was carried out to determine an SDF cut-off. There was an inverse relationship between SDF in the ejaculate of the diagnostic semen sample and CP (p = 0.02; OR 0.94 95% CI (0.90, 0.989)) as well as LB (p = 0.04; OR 0.95 95% CI (0.90, 0.9985)). No significant association was found between SDF after gradient and IUI outcome in the diagnostic sample nor between SDF (ejaculate/after gradient) in the IUI samples. The ROC analysis proposed a cutoff of 17.5% as the best compromise between sensitivity and specificity in the diagnostic SDF for live birth; however, the test diagnostics are low, with an AUC of 0.576. Overall, this study strengthens the hypothesis of an inverse relationship between SDF and CP/LB. Furthermore, SDF taken together with other clinical characteristics might provide more insight into male reproductive potential and predicting IUI outcome. Couples with SDF ≥ 17.5% in the diagnostic semen sample did not reach live birth. Further research is necessary to establish the diagnostic and prognostic potential of SDF as an add-on test.