Sperm DNA Fragmentation Index Research Articles

Single sperm vitrification with permeable cryoprotectant-free medium is more effective in patients with severe oligozoospermia and azoospermia

Testicular sperm extraction (TESE) is an invasive surgery for achieving the spermatozoa in cases with azoospermia. In these patients, the number of retrieved spermatozoa is limited and the optimal cryo-storage is very critical for their fertility preservation. Therefore, single sperm vitrification has been introduced for preservation of low number of spermatozoa.The goal was to assess the efficacy of sperm freezing medium (SFM) and sucrose medium as cryoprotectants for single sperm vitrification in cases with severe oligozoospermia and azoospermia.A total of 20 ejaculates from severe oligozoospermia and 20 testicular samples from azoospermia were processed. Twenty-five sperm cells were collected using ICSI injection pipette and transferred to a cryoprotectant droplet placed on the Cryotech, then vitrified by plunging in liquid nitrogen. Sperm motility, viability, fine-morphology, mitochondrial activity and DNA fragmentation index (DFI) were assessed before and after vitrification.Sperm motility, viability and the percentage of cells with mitochondrial activity were significantly decreased after vitrification in both severe oligozoospermic and testicular samples in either cryoprotectants. However, the rates of post-warm sperm motility and the cells with mitochondrial activity increased significantly in sucrose medium in both severe oligozoospermic and testicular samples compared to SFM. In testicular samples, the DFI of spermatozoa vitrified in SFM was significantly higher than those vitrified with sucrose medium.Sperm motility, viability, mitochondrial activity, and DNA integrity were better preserved in sucrose medium than SFM after single cell vitrification. The presented method may be a useful candidate for successful freezing of individual sperm cells in clinical setting.

Rapid cryopreservation of small quantities of human spermatozoa by a self-prepared cryoprotectant without animal component.

Cryopreservation of small quantities of human spermatozoa whilst maintaining adequate post‐thawing motility has been found an essential challenge for male fertility preservation. Therefore, the study used an effective, and convenient rapid‐freezing method to freeze small amounts of human spermatozoa by adding self‐prepared cryoprotectant (SPC) without animal component. In the feasibility experiment, no significant differences in progressive motility, normal sperm morphology, vitality or DNA fragmentation index between the conventional slow freezing and rapid freezing were realised. The present study prospectively analysed the effects of sperm freezing and resuscitation in 175 patients with severe oligozoospermia (sperm concentration <1 × 106/ml). We observed the 120 severe oligozoospermia specimens had a mean recovery rate of 60.19% ± 10.43% and a mean cryosurvival rate of 68.0% ± 9.17%. In addition, 55 cryptozoospermia specimens were analysed. The small‐volume cryopreservation showed advantages. The total sperm recovery, motility recovery and sperm loss rates were 98.48%, 50.17% and 1.52% respectively. In short, the SPC is safe and effective, and can be used to rapidly freeze severe oligozoospermia specimens. That is useful for successful sperm freezing whilst avoiding the risk of azoospermia in the later stages and promoting comprehensive fertility preservation.

Metabolic enzyme gene polymorphisms predict the effects of antioxidant treatment on idiopathic male infertility.

To explore the relationship between genetic polymorphisms of metabolic enzymes such as CYP1A1, CYP2D6, GSTM1, GSTT1, and GSTP1 and idiopathic male infertility. By observing the efficacy of antioxidants in the treatment of idiopathic male infertility, the effect of metabolic enzyme gene polymorphisms on antioxidant therapy in patients with idiopathic male infertility was prospectively studied. This case–control study included 310 men with idiopathic infertility and 170 healthy controls. The cytochrome P450 1A1 (CYP1A1), cytochrome P450 2D6 (CYP2D6), glutathione S-transferase M1 (GSTM1), glutathione S-transferase T1 (GSTT1), and glutathione S-transferase P1 (GSTP1) genotypes in peripheral blood samples were analyzed by polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP). The idiopathic male infertility group was treated with vitamin C, vitamin E, and coenzyme Q10 for 3 months and followed up for 6 months. GSTM1(−), GSTT1(−), and GSTM1/T1(−/−) in the idiopathic male infertility groups were more common than those in the control group. The sperm concentration, motility, viability, mitochondrial membrane potential (MMP), and seminal plasma total antioxidant capacity (T-AOC) level in patients with GSTM1(−), GSTT1(−), and GSTM1/T1(−/−) were lower than those in wild-type carriers, and the sperm DNA fragmentation index (DFI), 8-hydroxy-2’-deoxyguanosine (8-OH-dG), and malondialdehyde (MDA) and nitric oxide (NO) levels were higher. Therefore, oxidative damage may play an important role in the occurrence and development of idiopathic male infertility, but antioxidant therapy is not effective in male infertility patients with GSTM1 and GSTT1 gene deletions.

Evaluating the effect of BESTFertil antioxidant complex on semen parameters and severity of asthenic syndrome in men with a recent history of novel coronavirus infection (COVID-19)

Introduction. Direct influence of novel coronavirus infection (COVID-19) on male fertility is still unproven. However, through prolonged fever, hypoxia, systemic inflammation and other yet incompletely described factors this disease may apparently cause not only “long COVID” syndrome, but also transitory non-specific impairment of spermatogenesis. The study objective is to evaluate the effectiveness of administration of Russian complex with unique composition BESTFertil in case of decreased fertility and asthenic syndrome in males after new coronavirus infection.Materials and methods. We performed a non-blinded randomized prospective controlled trial which included 60 male patients with a history of COVID-19 not earlier than 6 months prior to enrollment from couples planning to have children. Patients were included if they had pre-disease semen analysis results available. They were divided into two equal groups. Group 1 (main group) received BESTFertil complex for 12 weeks, while Group 2 (comparison group) received no additional treatment. At baseline and 12-week follow-up all patients underwent semen analysis, MAR test, sperm DNA fragmentation test, serum sex hormone profiling and completed the “Asthenic condition scale” questionnaire. Student's t-test, Wilcoxon test, Mann-Whitney U-test and McNemar test were used for statistical analysis.Results. When comparing retrospective and baseline values we found no critical influence of COVID-19 on semen parameters, though a moderate decrease of percentage of sperm with progressive motility could be observed (40.5 % vs 30 %, p = 0.008). Group 1 had a stronger tendency toward recovery of semen parameters. Patients receiving BESTFertil had median total motile sperm count increased from 48.5 million to 76.8 million (p = 0.032), while in comparison group it increased from 39.8 million to 49.2 million (p = 0.317). BESTFertil antioxidant complex had no negative influence on endocrine profile. Moreover, there was a statistically significant decrease of sperm DNA fragmentation index from 21.2 to 13.9 % (p = 0.007) in Group 1. Lower rate of asthenic symptoms was observed in Group 1 which received BESTFertil.Conclusion. Men with a recent history of COVID-19 had a moderate decrease of surrogate fertility markers, most probably due to non-specific mechanisms. Patients receiving BESTFertil had a more pronounced recovery of semen parameters and improvement in post-COVID asthenic syndrome.

The effect of age and abstinence time on semen quality: a retrospective study.

This study analyzed the effects of male age and abstinence time on semen quality and explored the best abstinence time for Chinese males among different age groups. Semen parameters, including sperm kinetics, morphology, and DNA fragmentation index (DFI), were reviewed from 2952 men. Samples were divided into six age groups (≤25 years, 26–30 years, 31–35 years, 36–40 years, 41–45 years, and >45 years) and were divided into six groups according to different abstinence time (2 days, 3 days, 4 days, 5 days, 6 days, and 7 days). The differences in semen quality between the groups were compared, and the effect of age and abstinence time on semen quality was analyzed. Significant differences were observed in semen volume, progressive motility (PR), and DFI among the age groups (all P < 0.05), and no significant differences were observed in sperm morphological parameters (all P > 0.05). There were significant differences in semen volume, PR, and DFI among different abstinence time groups (all P < 0.05) and no significant differences in sperm morphological parameters (all P > 0.05). Pearson analysis showed that male age and abstinence time were both significantly correlated with sperm kinetics and DFI (both P < 0.05), while no significant correlation was found with sperm morphological parameters (all P > 0.05). The box plots and histograms of men's age, abstinence time, and semen quality show that most semen quality parameters differ significantly between the 2 days and 7 days abstinence groups and other groups at different ages. Except for the sperm morphology parameters, sperm kinetic parameters and sperm DFI are linearly related to male age and abstinence time.

Magnetic-activated cell sorting of nonapoptotic spermatozoa with a high DNA fragmentation index improves the live birth rate and decreases transfer cycles of IVF/ICSI.

The present study aimed to evaluate the clinical outcomes of magnetic-activated cell sorting (MACS) in sperm preparation for male subjects with a sperm DNA fragmentation index (DFI) ≥30%. A total of 86 patients who had undergone their first long-term long protocol were selected. The protocol involved in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) cycles, and the patients were divided into the MACS or control groups. The MACS group included sperm samples analyzed with MACS that were combined with density gradient centrifugation (DGC) and the swim-up (SU) technique (n = 39), and the control group included sperm samples prepared using standard techniques (DGC and SU; n = 41). No differences were noted with regard to basic clinical characteristics, number of oocytes retrieved, normal fertilization rate, cleavage rate, or transplantable embryo rate between the two groups in IVF/ICSI. In addition, the clinical pregnancy and implantation rates of the first embryo transfer cycles indicated no significant differences between the two groups. However, there was a tendency to improve the live birth rate (LBR) of the first embryo transfer cycle (63.2% vs 53.9%) and the cumulative LBR (79.5% vs 70.7%) in the MACS group compared with the control group. Moreover, the number of transferred embryos (mean ± standard deviation [s.d.]: 1.7 ± 0.7 vs 2.3 ± 1.6) and the transfer number of each retrieved cycle (mean ± s.d.: 1.2 ± 0.5 vs 1.6 ± 0.8) were significantly lower in the MACS group than those in the control group. Thus, the selection of nonapoptotic spermatozoa by MACS for higher sperm DFI could improve assisted reproductive clinical outcomes.

Vitamin D3 Supplementation Effects on Spermatogram and Oxidative Stress Biomarkers in Asthenozoospermia Infertile Men: a Randomized, Triple-Blind, Placebo-Controlled Clinical Trial.

The objective is to evaluate the effects of vitamin D3 (VD3) on sperm parameters and seminal and serum oxidative stress (OS) biomarkers in asthenozoospermia infertile men. This randomized, triple-masking, placebo-controlled clinical trial conducted on 86 asthenozoospermia infertile men with serum 25 hydroxy vitamin D3 (25-OH-D3) < 30ng/ml in the infertility clinic of Ahvaz Jahad Daneshgahi, Iran. Patients were randomly allocated to groups A and B, who received daily 4000IU vitamin D3 (VD3) and matching placebo respectively for 3months. Demographic data, dietary intake, physical activity, sun exposure, anthropometric indices, serum and seminal levels of MDA (Malondialdehyde), 8-hydroxy-2- Dioxy Guanosine (8-OHDG), total antioxidant capacity (TAC) and calcium, sperm DNA fragmentation index (DFI), serum 25-OH-D3, parathyroid hormone (PTH), phosphorus, and sperm parameters were assessed. VD3 supplementation had no significant effects on body weight, body mass index (BMI), waist circumference (WC), body fat (BF), 8-OHDG, DFI, semen volume, sperm count, and normal sperm morphology, but increased post-intervention mean and mean change of serum 25-OH-D3 (P < 0.001, P < 0.001), PTH (P < 0.001, P < 0.001) and phosphorus (P = 0.009, P = 0.049) and seminal calcium (P = 0.035, P = 0.038) and serum calcium (P = 0.008, P = 0.009), seminal TAC (P < 0.001, P < 0.001), and serum TAC (P = 0.007, P = 005), total sperm motility (P < 0.001, P < 0.001) and progressive sperm motility (P < 0.001, P < 0.001) and decreased seminal MDA (P = 0.017, P = 0.004) and serum MDA (P = 006, P = 0.005) significantly compared to the baseline and placebo group respectively. VD3 supplementation may modulate OS and affect sperm motility in men with asthenozoospermia and serum 25-OH-D3 < 30ng/ml. Iran Clinical Trials Registry, ID: IRCT20151128025274N4, registered on 28 March 2018, URL of trial registry record: https://www.irct.ir/trial/29983.

Morphokinetic parameter comparison between embryos from couples with high or low sperm DNA fragmentation index

To study whether time-lapse imaging can identify morphokinetic events impacted by a high sperm DNA fragmentation index (DFI). Historical cohort study. Private university-affiliated invitro fertilization center. A total of 978 zygotes cultured until day 5 in a time-lapse imaging incubator between March 2019 and August 2020, derived from 118 patients undergoing intracytoplasmic sperm injection as a result of idiopathic male factor infertility. Kinetic markers from the point of insemination were recorded. Generalized linear mixed models adjusted for potential confounders followed by the Bonferroni post hoc test were used to compare the timing of specific events in patients with a low (<30%) or high (≥30%) sperm DFI. The recorded kinetic markers were the following: timing to pronuclei appearance and fading; timing to 2, 3, 4, 5, 6, 7, and 8 cells; and timing to start blastulation and blastulation. Timing to blastulation. Embryos derived from sperm samples with ≥30% DFI showed significantly slower divisions compared with those with <30% DFI (mean differences of 0.7 hours in timing to pronuclei appearance, 1.2 hours in timing to pronuclei fading, 1.5 hours in timing to 2 cells, 2.5 hours in timing to 3 cells, 1.8 hours in timing to 4 cells, 3.3 hours in timing to 5 cells, 3.1 hours in timing to 6 cells, 3.2 hours in timing to 7 cells, 2.7 hours in timing to 8 cells, 8.4 hours in timing to start blastulation, and 3.8 hours in timing to blastulation). The incidences of reverse or direct cleavages (9.3% vs. 4.4%; odds ratio [OR], 2.24; 95% confidence interval [CI], 1.32-3.77) and multinucleation at 2-cell (18.9% vs. 12.0%; OR, 1.70; 95% CI, 1.12-2.58) and 4-cell (14.2% vs. 6.4%; OR, 2.42; 95% CI, 1.57-3.74) stages were significantly higher in embryos deriving from ≥30% DFI than from <30% DFI. The KIDScore ranked significantly different between embryos derived from samples with <30% and ≥30% DFI. Continuous DFI was positively correlated with all timings of specific events and with the incidences of abnormal cleavage patterns (OR, 1.042; 95% CI, 1.025-1.059) and multinucleation at 2-cell stage (OR, 1.053; 95% CI, 1.030-1.076) and inversely correlated with the KIDScore rank (B, -0.218; 95% CI, -0.044 to -0.007). No significant differences were observed in clinical outcomes between the groups. Embryo morphokinetic parameters are negatively impacted by high sperm DFI, resulting in delayed cell cleavage and blastulation.

SPERM DNA FRAGMENTATION AND THE RELATIONSHIP WITH SEMEN PARAMETERS

Objective: To evaluate the level of sperm DNA fragmentation and its association with semen parameters. Material and methods: this cross-sectional study recruited 289 semen samples from the husband of infertile couples who were examined at the Center for Reproductive Endocrinology &amp; Infertility, Hue University of Medicine and Pharmacy Hospital. Evaluated values include semen parameters, sperm DNA fragmentation index (DFI). Results: The mean of DFI was 28.37 ± 20.38 (4 - 91)%, the rate of DFI &lt; 30% group was 64.0% and DFI ≥ 30% was 36.0%: Progressive motility in DFI &lt;30% group was higher than DFI ≥ 30% group, statistically significant, (29.87 ± 11.33 vs 26.85 ± 12.61, p = 0.038). DFI was positive correlated with progressive motility (r = - 0.132, p = 0.025), and tends to be positively correlated with the abnormality head sperm (r = 0.111, p = 0.061). Conclusion: High levels of sperm DNA fragmentation are observed in infertile couples. The sperm DNA fragmentation index is not strongly correlated with semen parameters. Sperm DNA fragmentation test should be performed as an additional step in evaluating male fertility Key words: semen parameters, sperm, sperm DNA fragmentation index - DFI, big halo

Effect of Sperm Dna Fragmentation Index on Clinical Outcomes of Intra-Uterine Insemination Patients

Objective: To study the effect of sperm chromatin condensation (DNA fragmentation index (DFI)) using aniline blue-eosin (AB-E) staining on pregnancy outcomes in patients facing unexplained infertility undergoing intra- uterine insemination (IUI). Our initial hypothesis states that if DNA fragmentation is high then chances of pregnancy are low/NIL hence these patients should be recommended with advanced ART procedures like IVF and ICSI. Design: Prospective study Setting: Tertiary care infertility centre Method: A total of 185 patients with age less than 40 years, non-smokers and without history of any pathogenic infection in the past 2months facing unexplained infertility i.e., males with normal semen analysis reports and females with normal ovulation and hysterosalpingography (HSG) reports were selected for the study. Patients were undergoing their first or second IUI treatment cycle between the period of June 2016 to December 2016. DNA fragmentation index (DFI) using aniline blue- eosin staining method was studied in semen samples provided on the day of IUI procedure. The patients were separated into 3 groups: low DFI (DFI&lt;= 10%), medium DFI (DFI=11 % - 20%), and high DFI (DFI &gt;= 21%) and clinical pregnancy outcomes of IUI were recorded. Statistical analysis was performed using Pearson correlation co-efficient, ANOVA and Shapiro Wilk Test on the above groups. Main Outcome Measures: DNA fragmentation index (DFI) (%), Clinical pregnancy rate (%) Result: The overall clinical pregnancy rate for the selected patient pool was 21.08% with an average DFI of 8.84% in the pregnant female group and 14.65% in the non-pregnant female group. Sperm DFI % and clinical outcomes in IUI treated patients were statistically significant and negatively correlated with correlation coefficient (r) of -0.1, -0.3 and -0.3 in low DFI%, medium DFI and high DFI% groups respectively. Conclusion: Our study demonstrated that DFI (%) and clinical pregnancy rate (%) are significantly and negatively correlated in patients with normal semen parameters undergoing IUI. The higher the DFI% the chances of clinical pregnancy become very low, therefore, these patients should not be recommended IUI but with advanced ART procedures like IVF and ICSI.

Correlation between Sperm Dna Fragmentation and Conventional Semen Parameters among Different Age Groups

Objective: To study the relationship between conventional semen parameters and sperm chromatin condensation (DNA fragmentation index) using aniline blue-eosin staining method among patients of different age groups visiting the In-vitro fertilization (IVF) clinic.Design: Retrospective study Setting: Tertiary care infertility centre Method: A total of 240 patient semen samples were studied between the period of May 2015 to May 2016 for conventional semen parameters (WHO criteria) and DNA fragmentation index (DFI) using aniline blue- eosin staining method. Patients were separated into three groups: &lt;=30 years, 31-35 years and 36 years &amp; above. Statistical analysis was performed using Pearson correlation co-efficient and regression tests on the groups. Main Outcome Measures: Sperm concentration (Millions /ml), motility(%), normal morphology(%), DFI (%). Result: In each age group, i.e., &lt;=30years, 31-35 years and 36 years &amp; above, there was a significant and negative correlation between DFI and sperm concentration (r= -0.50, r= -0.34, r= -0.49 respectively; P&lt;0.05), motility(r= -0.69,r= -0.66, r= -0.54 respectively; P&lt;0.05) and normal morphology (r= -0.86,r= -0.80, r= -0.75 respectively; P&lt;0.05). Sperm DNA fragmentation index among the age groups was not statistically significantly (P&gt;0.05). Conclusion: Our study demonstrated that age is not a predictor of DFI. Whereas, sperm concentration, sperm motility and normal sperm morphology showed a significant association with DFI in all the age groups i.e., better the conventional semen parameters, lower the DFI.

Comprehensive analysis of male ejaculate from infertile couples in the age aspect

Introduction. According to Russian Association of Urology a number of infertile couples ranges from 8 to 17.2 % in various regions of Russia. One of the causes of which is the male factor. Raising the age of the father reduces the chance of conception, increases the risk of early termination of pregnancy and the risk of certain diseases of the child. In this regard the existing knowledge of age-related changes in ejaculate parameters needs to be de veloped. This is particularly true for men of infertile couples.The study objective – to investigate the age patterns of change in the spermological status of infertile men and identify the most age-related variations.Materials and methods. Research design – observation, retrospective, crossection. Analysis of the medical records of 517 Novosibirsk male residents from couples with infertility in marriage was made. The sampling is divided into 5 age groups: 1st group (n = 108) – men of ≤29.9 years old; 2nd group (n = 171) – 30.0–34.9 years old; 3rd group (n = 130) – 35.0–39.9 years old; 4th group (n = 68) – 40.0–44.9 years old; 5th group (n = 40) – ≥45.0 years old.Results. Median of male age in total sample – 34 years, frequency of persons over 40 years – 21 %. Overall sample shows a decline in reference values of proportion of morphologically normal forms – 3.0 (1.5–5.0) % and increase in sperm DNA fragmentation index (SDFI) – 16.0 (11.0–22.2) %. In groups from 1st to 5th there is an age-specific decrease in ejaculate volume (3.5 (2.5–4.8) ml; 3.3 (2.4–4.2) ml; 3.2 (2.4–4.5) ml; 3.0 (1.7–4.9) ml; 2.7 (1.9–3.5) ml, р1–5 &lt;0.005) and sperm count (127.1 (58.8–264.0) million; 122.5 (62.4–214.0) million; 118.5 (52.8–217.5) million; 98.0 (58.5–199.5) million; 81.0 (36.5–137.9) million, р1–5, 2–5 &lt;0.005). SDFI has an age-associated increase in the value of the indicator (14.4 (10.8–19.5) %; 15.0 (10.0–21.0) %; 17.2 (11.0–22.0) %; 18.5 (14.0–24.2) %; 19.2 (13.2–29.6) %, р1–4 &lt;0.005).Conclusion. It was first time shown for the male population of Novosibirsk of Siberian Federal District that a reduction in the proportion of morphologically normal sperm below the reference values observed in the total sample and it did not depend on the age of the men examined. Only three of the examined indicators of sperm status studied infertile males had a significant age dependency: ejaculate volume decrease, reduction of total sperm, increased DNA fragmentation of sperm. The lack of a reliable influence of age on HBA-test indicators was first shown. However the trends identified require further study. The results show that age-associated increase in DNA fragmentation of sperm starts with young men under 30, and underscores the need to define SDFI in actual clinical practice. And negative processes of age influence on the quantitative, morphological and ultrastructural characteristics of sperm occur most frequently in men after 40 years.

Improvement roles of zinc supplementation in low dose lead induced testicular damage and glycolytic inhibition in mice

Lead (Pb) is a toxic metal that affects the male reproductive system. This study aimed to investigate the effects of zinc (Zn) intake between recommended dietary allowances (RDAs) and tolerable upper intake levels (ULs) in preventing male testis damage induced by low-dose Pb. Forty-five mice were randomly divided into control, Pb, and Pb + Zn groups. They were given distilled water ad libitum with 0, 200 mg/L Pb2+, or 15 mg/L Zn2+ mixed with 200 mg/L Pb2+ for 90 consecutive days. The Zn levels in the blood and testis of the Pb group were significantly lower than those of the control group. The Pb levels in the blood and testis of the Pb + Zn group were significantly lower than those of the Pb group. Additionally, a significant decrease in sperm density and viability, with a significant increase in sperm abnormality rate and DNA fragmentation index, was observed in the Pb group. Zn supplementation significantly improved the above sperm parameters. Moreover, Zn supplementation decreased low-dose Pb-induced lipid peroxidation and increased glutathione, total superoxide dismutase (SOD), and copper/Zn-SOD levels. Furthermore, Zn treatment improved glycolysis products and lactate transporters in Pb-treated mouse testes. Our findings suggest that Zn intake between RDAs and UL can act as a therapeutic agent in protecting against the reproductive impairments associated with Pb exposure.

PD29-09 TRIGLYCERIDE-GLUCOSE INDEX (TYG) IS ASSOCIATED WITH SPERM PARAMETERS AND SPERM DNA FRAGMENTATION IN PRIMARY INFERTILE MEN: A CROSS-SECTIONAL STUDY

You have accessJournal of UrologyInfertility: Epidemiology & Evaluation II (PD29)1 Sep 2021PD29-09 TRIGLYCERIDE-GLUCOSE INDEX (TYG) IS ASSOCIATED WITH SPERM PARAMETERS AND SPERM DNA FRAGMENTATION IN PRIMARY INFERTILE MEN: A CROSS-SECTIONAL STUDY Luca Boeri, Paolo Capogrosso, Edoardo Pozzi, Luigi Candela, Federico Belladelli, Walter Cazzaniga, Giuseppe Fallara, Nicolò Schifano, Eugenio Ventimiglia, Daniele Cignoli, Gianmarco Colandrea, Massimo Alfano, Costantino Abbate, Francesco Montorsi, and Andrea Salonia Luca BoeriLuca Boeri More articles by this author , Paolo CapogrossoPaolo Capogrosso More articles by this author , Edoardo PozziEdoardo Pozzi More articles by this author , Luigi CandelaLuigi Candela More articles by this author , Federico BelladelliFederico Belladelli More articles by this author , Walter CazzanigaWalter Cazzaniga More articles by this author , Giuseppe FallaraGiuseppe Fallara More articles by this author , Nicolò SchifanoNicolò Schifano More articles by this author , Eugenio VentimigliaEugenio Ventimiglia More articles by this author , Daniele CignoliDaniele Cignoli More articles by this author , Gianmarco ColandreaGianmarco Colandrea More articles by this author , Massimo AlfanoMassimo Alfano More articles by this author , Costantino AbbateCostantino Abbate More articles by this author , Francesco MontorsiFrancesco Montorsi More articles by this author , and Andrea SaloniaAndrea Salonia More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002030.09AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Male factor infertility (MFI) is often associated with metabolic disorders such as diabetes mellitus and metabolic syndrome, where insulin resistance (IR) plays a relevant pathological role. The Homeostatic Model Assessment (HOMA) and triglycerides/glucose index (TyG) are a good marker of IR. We aimed to investigate the relationship between the TyG and both semen and hormonal characteristics in a cohort of primary infertile men. METHODS: Complete demographic, clinical and laboratory data from 726 consecutive primary infertile men (infertility defined according to the WHO definition) were considered. Health-significant comorbidities were scored with the Charlson Comorbidity Index (CCI). Serum hormone, semen analyses (based on 2010 WHO reference criteria) and sperm DNA fragmentation index (DFI) were measured in every patient. Glucose, insulin and triglycerides levels were measured after a 12-h overnight fast, and HOMA-IR and TyG indices were calculated and classified using a 2.6 threshold (as from Ascaso et al. 2003) and a 8.1 threshold (as from Kang et al. 2017), respectively. Descriptive statistics and logistic regression models tested the association between TyG with hormonal milieu and semen parameters. RESULTS: Median (IQR) age was 39 (35-43) years. A TyG and HOMA suggestive for IR was found in 339 (46.6%) and 154 (21.2%) patients, respectively and 121 (16.7%) had both. At Spearman’s test, TyG was correlated with HOMA-IR score (rho = 0.46, p<0.001). Compared to men with normal TyG, men with TyG>8.1 were older [37 (34-41) vs. 39 (35-43) years], had greater BMI [24.5 (23-27) vs. 26 (24-28) kg/m2] and CCI scores (0 vs 1), lower total testosterone [4.5 (3.4-5.6) vs. 4.2 (3.1-5.5)] and sperm concentration [8.0 (1.5-25.0) vs. 12 (2.8-30)], but higher DFI [38.2 (23-54) vs. 28.0 (18-48)] and presented a greater proportion of NOA (24.8% vs. 16.3%) (all p<0.01). At multivariable logistic regression analysis, men with TyG>8.1 were at higher risk of NOA [OR 1.78 (CI: 1.1-2.8)] and DFI>30 [OR 1.92 (CI: 1.2-2.9)], after accounting for patient age, CCI, FSH values, and testicular volume. CONCLUSIONS: Patients with TyG suggestive for IR showed worse clinical, hormonal and semen parameters compared to those with lower TyG values. TyG index represents a reliable marker of IR for primary infertile men in the real-life setting, thus supporting an easier long-term follow-up of patients at higher risk of chronic dysmetabolic comorbidities. Source of Funding: None © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e530-e530 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Luca Boeri More articles by this author Paolo Capogrosso More articles by this author Edoardo Pozzi More articles by this author Luigi Candela More articles by this author Federico Belladelli More articles by this author Walter Cazzaniga More articles by this author Giuseppe Fallara More articles by this author Nicolò Schifano More articles by this author Eugenio Ventimiglia More articles by this author Daniele Cignoli More articles by this author Gianmarco Colandrea More articles by this author Massimo Alfano More articles by this author Costantino Abbate More articles by this author Francesco Montorsi More articles by this author Andrea Salonia More articles by this author Expand All Advertisement Loading ...

SPERM DNA FRAGMENTATION TEST IN ADDITION TO GENERAL SEMEN TEST MAY LEAD TO ELUCIDATION OF POTENTIAL FACTORS FOR PREDICTING IVF OUTCOMES

Female factors are very important in determining the success of in vitro fertilization (IVF). However, many reports have shown that male factors also affect the success or failure of IVF. Therefore, the sperm evaluation is important, semen examination criteria based on conventional methods alone is not a primary factor of consideration for determining IVF embryonic potential, pregnancy rates, and miscarriage rates after transplantation. We performed and analyzed the sperm DNA fragmentation index (DFI) and general semen test based on the WHO criteria and compared the two using semen factors. This study was conducted from June 2020 to December 2020. A total of 134 male patients with the main complaint of infertility who requested semen testing were included. We evaluate sperm motility using an automatic sperm analyzer based on the WHO criteria and sperm morphology via the Kruger test (general semen tests). DFI and general semen test were conducted in accordance with the WHO criteria, and correlations between the two tests were investigated. The WHO criteria were set as the cutoff value for each of the following factors: fluid volume, concentration, motility, and normal morphology, and compared with the DFI results. The mean age of the 134 patients at the time of the sperm tests was 38.8±6.0 years (range=26-52 years). The subjects had a mean DFI of 15.6±11.5% and the DFI increased with age (r=0.2319, p<0.01). In contrast, the concentration, motility, and normal morphology decreased as the DFI increased (concentration; r=-0.1992, p<0.05; motility; r=-0.2672, p<0.01; normal morphology; r=-0.2881, p<0.001). Patients who satisfied the WHO criteria in terms of sperm concentration, motility, and normal morphology had a significantly lower DFI than the group that did not satisfy the criteria (concentration p<0.05; motility p<0.01; normal morphology p<0.01). Therefore, evaluation with a general semen test based on the WHO criteria should be regarded as a qualitative evaluation of all factors other than fluid volume. Male infertility due to DFI should be suspected when in vitro fertilization results are poor despite normal semen findings based on the WHO criteria. The results of this study suggest that the DFI test may be an important evaluation in follow-up semen analyses.

Low-dose Methotrexate Therapy Does Not Affect Semen Parameters and Sperm DNA.

Methotrexate is widely used in inflammatory diseases during the patients' reproductive years. The effect on male fertility and sperm DNA integrity is largely unknown. We evaluated sperm DNA integrity and basic semen parameters according to the World Health Organization (WHO) in male patients with inflammatory diseases treated with methotrexate. Semen samples from 14 patients on low-dose maintenance methotrexate were compared with samples from 40 healthy volunteers. Further, 5 patients delivered samples on and off methotrexate therapy for paired comparison. Sperm DNA fragmentation index (DFI), concentration, motility, and morphology were evaluated. Blood sex hormones and methotrexate levels were measured in blood and semen. DNA fragmentation index in methotrexate-treated patients was comparable with that in healthy volunteers (DFI, 11.5 vs 15.0; P = .06), and DFI did not change significantly on and off methotrexate in the paired samples (DFI, 12.0 vs 14.0; P = 0.35). Sperm concentration, motility, and morphology did not differ between men treated with methotrexate and healthy volunteers. Sperm progressive motility increased off therapy compared with on therapy (65.0% vs 45.0%, P = .04), but all fluctuations in progressive motility were within the WHO reference interval. All methotrexate polyglutamates1-5 were detected in blood, but only methotrexate polyglutamate1 in semen. Serum testosterone was unaffected by methotrexate therapy. Patients treated with low-dose methotrexate have a sperm quality comparable with that of healthy volunteers, and methotrexate treatment does not increase sperm DNA fragmentation. This study does not support cryopreservation of semen before treatment initiation nor a 3-month methotrexate-free interval prior to conception.

Sperm DNA fragmentation index and cumulative live birth rate in a cohort of 2,713 couples undergoing assisted reproduction treatment

To study how the choice of the first assisted reproductive technology treatment type affects the cumulative live birth rate (CLBR) in couples with high sperm DNA fragmentation index (DFI). Longitudinal cohort study. University-affiliated fertility clinic. A total of 2,713 infertile couples who underwent assisted reproductive technology treatment between 2007 and 2017 were included in the study. All invitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) treatments (up to three fresh treatments and all associated frozen-thawed embryo transfers) offered to the couples by the public health care system were included, in total 5,422 cycles. None. The primary outcome was the CLBR. The secondary outcomes were the fertilization rate and the miscarriage rate. The IVF and ICSI groups were defined according to the method applied in the first treatment cycle. In the IVF group, the CLBR values were higher for couples with normal DFI compared with those for couples with high DFI (≥20%) (48.1% vs. 41.6% for conservative CLBR estimate and 55.6% vs. 51.4% for optimal CLBR estimate after adjustment for female age, respectively). No DFI-dependent difference was seen in the ICSI group. Our results demonstrated that a high DFI predicts a statistically significantly lower CLBR if IVF and not ICSI is applied in the first cycle of assisted reproduction.

O-209 The impact of sperm DNA fragmentation on pregnancy outcomes depends on oocyte dimorphisms

Abstract Study question Does the impact of sperm DNA fragmentation (SDF) on Intracytoplasmic sperm injection (ICSI) outcomes depend on the presence of oocyte dimorphisms? Summary answer There is a significant influence of oocyte quality on the impact of SDF on pregnancy outcomes. What is known already Sperm DNA fragmentation has been associated with ICSI outcomes. DNA damage is commonly encountered in human spermatozoa and it has been widely accepted that the oocyte assumes responsibility for the repair and remodelling of both the maternal and paternal genomes during the oocyte-embryo transition. Indeed, spermatozoa with DNA damage can fertilise oocytes and still lead to embryo development due to the oocyte DNA repair capacity. Considering the vital role played by the oocyte in the developmental process, it could be hypostatised that the oocyte quality, translated as oocyte morphology, would influence the machinery responsible for sperm DNA repair after fertilization. Study design, size, duration This cohort study included 3,035 oocytes from 525 patients undergoing ICSI cycles in a university-affiliated IVF-center, between June/2016 and July/2019. Oocytes were split into groups according to the SDF index of the sample used for ICSI: low-fragmentation (&amp;lt;30% SDF, n = 2,277) and high-fragmentation (≥30% SDF, n = 758). Oocytes were evaluated before sperm injection and the dimorphisms were recorded. The influence of SDF index on ICSI outcomes, depending on the presence of oocytes dimorphisms was evaluated. Participants/materials, setting, methods Data was evaluated using generalized linear models (GZLM) followed by Bonferroni post hoc. The results are expressed as mean ± standard error for continuous variables or percentages for dichotomous variables, and p-values. The sample size calculation suggested that a sample of at least 504 subjects had 95% power to detect a 20% effect with a significance level of 5% (α). The study was performed in a private university–affiliated in vitro fertilization (IVF) center. Main results and the role of chance The association of both factors: the presence of oocyte dimorphisms (dark cytoplasm, vacuoles in the ooplasm, and resistant membrane) and high SDF index resulted in the lowest fertilization rate among groups, while oocytes free of these dimorphisms injected with samples with &amp;lt;30% SDF had the highest fertilization rate (p = 0.05, p &amp;lt; 0.01 and p &amp;lt; 0.01 for dark cytoplasm, vacuoles in the ooplasm and resistant membrane respectively). The impact of SDF index on high quality embryos rate on cleavage stage was also influence by the presence smooth endoplasmic reticulum clusters and resistant membrane oocytes (p = 0.013 and p = 0.018). As for the clinical outcomes, the impact of SDF index on the implantation rate was influenced by the presence of vacuoles in the ooplasm (p &amp;lt; 0.01), smooth endoplasmic reticulum clusters (p &amp;lt; 0.01), large perivitelline space (p &amp;lt; 0.01), resistant membrane (p &amp;lt; 0.01), and non-resistant membrane (p &amp;lt; 0.01), while the influence of SDF index on the pregnancy rate was influenced by the presence large perivitelline space (p &amp;lt; 0.01), resistant membrane (p = 0.018) and non-resistant membrane (p &amp;lt; 0.01). The effect of SDF on the miscarriage rate was also increased in the presence of large perivitelline space (p = 0.045), non-resistant membrane (0.037) and centrally located cytoplasmic granular area (p = 0.025). Limitations, reasons for caution The retrospective nature is a limitation. It could be argued that using samples with high SDF index does not necessarily mean that a sperm cell with a fragmented DNA was injected, however, the higher the SDF index, the higher the chance of selecting one with fragmented DNA. Wider implications of the findings The findings presented here highlight the crucial role of male and female factors when facing assisted reproduction. The association of low oocyte quality and high SDF index may lead to impaired results. As the oocyte defect cannot be modified, in vivo upgrading of spermatozoa before the treatment should be encouraged. Trial registration number Not applicable

O-151 Detailed characterization of infertile men with idiopathic versus unexplained infertility: a single-center experience

Abstract Study question We aimed to investigate the rate of and the clinical characteristics of men with idiopathic versus unexplained infertility from a cohort of white-European men. Summary answer Approximately 20% and 5% of men evaluated for primary couple’s infertility depicted characteristics suggestive for idiopathic and unexplained infertility, respectively. What is known already Male factor infertility (MFI) can be associated with clinical, hormonal and genetic diseases, but MFI is idiopathic in almost 30% of cases. Study design, size, duration Data from 3098 infertile men (according to WHO definition) consecutively evaluated between 2003-2020 at a single academic centre were analysed and compared with those of 103 fertile controls. Health-significant comorbidities were scored with the Charlson Comorbidity Index (CCI). Idiopathic infertility was defined for pathological semen analysis but normal physical examination and endocrine, genetic and biochemical laboratory testing. Unexplained infertility is defined as infertility of unknown origin with normal sperm parameters. Participants/materials, setting, methods Testicular volume (TV) was assessed with a Prader’s orchidometer. Serum hormones and sperm DNA fragmentation index (SDF) were measured in every patient. Vitamin D3 (VitD) deficiency was considered for vitD levels &amp;lt;20 ng/mL. Semen analyses were based on the 2010 WHO reference criteria. Health-significant comorbidities were scored with the Charlson Comorbidity Index (CCI). Main results and the role of chance Overall, 570 (18.5%) and 154 (5.0%) patients depicted criteria suggestive for either idiopathic or unexplained primary infertility, respectively. Groups were similar in terms of age, BMI, CCI, recreational habits, circulating serum hormones and SDF. Testicular volume was lower in men with idiopathic vs. unexplained infertility [median (IQR) 20 (15-25) vs. 20 (17-25); p &amp;lt; 0.001]; more idiopathic than unexplained infertile men depicted TV &amp;lt; 15ml (23.4% vs. 12%; p &amp;lt; 0.01). Similarly, vitD levels were lower [22 (17-28) vs. 27 (21-42) ng/mL; p &amp;lt; 0.001] in idiopathic vs. unexplained infertile men, with a higher rate of pathologic VitD levels in the same group (42.1% vs. 10%; p = 0.04). When compared to fertile controls, groups were similar in terms of age, BMI, CCI and serum hormones values. TV was larger in fertile controls than idiopathic and unexplained infertile men (all p &amp;lt; 0.01). At multivariable logistic regression analysis only vitD deficiency (OR 8.1, p = 0.03) was found to be associated with idiopathic infertility after accounting for age, BMI, testosterone values and TV. Limitations, reasons for caution The small number of fertile controls may raise the possibility of biases. Wider implications of the findings Idiopathic and unexplained infertility were identified in approximately 20% and 5% of men evaluated, respectively. Idiopathic infertile men showed lower TV and lower vitD values compared to men with unexplained infertility. Future studies are needed to develop a more tailored management to these difficult MFI cases. Trial registration number .

O-212 MACS vs TESA for raised sperm DNA fragmentation index – a RCT

Abstract Study question In Individuals with raised Sperm DNA Fragmentation Index (SDF), will sperm selection by magnetic activated cell sorting (MACS) or surgical retrieval of testicular sperms (TESA) optimize the reproductive outcomes? Summary answer Couples with failed implantation raised SDF, TESA /MACS offer similar results. This RCT doesn’t prove superiority or added benefit with any of the above interventions. What is known already It is evident that raised SDF negatively affects the reproductive outcomes. Management for raised SDF to optimize reproductive outcomes is still elusive. Study design, size, duration This was a Randomized Control Trial (RCT) with prior approval from institutional Ethical Committee and trial registration. Couples undergoing stimulation with raised SDF were randomized to MACS (n = 75) and TESA (n = 75) for sperm selection between April2019 &amp; February2020. Participants/materials, setting, methods Couples with history of one failed IVF had SDF testing and SDF&amp;gt;30% were recruited. SDF test done with SCSA method and randomized using software. ICSI was the method of insemination. Extended embryo culture till blastocyst was done and freeze all policy was opted. Two Blastocysts that showed 100% survival were transferred in a Frozen Embryo transfer (FET) cycle. Embryonic and Reproductive outcomes were compared between both groups. Live birth and Miscarriage were the primary outcomes. Main results and the role of chance Reproductive Outcomes of MACS Vs TESA were: Average Blastocyst conversion - 32% Vs 39% (RR 1.22, CI1.00 to 1.50) Implantation rate (IR) - 50% Vs 35% (RR - 0.71, CI 0.51 to 0.98) Miscarriage rate (MR) - 5.3% Vs 11% (RR1.6333, CI 0.5227 to 5.1039) Multiple Pregnancy rate (MPR) - 8% Vs 4% Live birth Rate (LBR) per Intention to treat (ITT) - 41.3% Vs 44% (RR 0.95, 95% CI 0.72 to 1.26) LBR per ET cycle - 63% Vs 56% (RR 1.23, 95% CI 0.77 to 1.94) Our preliminary results suggest that despite greater availability of blastocysts for transfer in the TESA group, no difference in ART outcomes was observed between the groups. Though the IR was statistically low with TESA, our primary outcomes LBR and MR were comparable. TESA or MACS seem to offer similar outcomes. Considering the invasiveness with TESA, MACS can be offered for better sperm selection for couples with raised sperm DFI &amp; failed implantation. Limitations, reasons for caution Small sample size. TESA is a surgical intervention Wider implications of the findings Optimal intervention for management of SDF still needs further research. Trial registration number CTRI/2019/07/020140