Sperm Deoxyribonucleic Acid Integrity Research Articles

Improved semen parameters and sperm deoxyribonucleic acid fragmentation following microscopic varicocelectomy: short term results of a prospective study

Background: Assessment of sperm deoxyribonucleic acid (DNA) damage post varicocelectomy, has brought about new prospects in treating men with infertility. Improvements in spermatogenesis with decrease in sperm DNA damage has been demonstrated which has led to significant enhancement in fertility rates. Therefore, we evaluated the efficacy of microscopic varicocelectomy on the reduction in sperm DNA damage in concurrence to the conventional improvement of semen parameters. Methods: All patients with a diagnosis of varicocele leading to infertility and planned for microscopic varicocelectomy from January 2013 through September 2019, were included in the study. Sperm chromatin structure assay (SCSA) was done preoperatively and at 3 and 6 months postoperatively for sperm DNA integrity assessment and the results expressed as sperm percentage DFI were compared. A p value <0.05 was considered statistically significant. Results: 105 infertile men underwent microscopic varicocelectomy. Mean age and infertility period from the date of unprotected sex were 26.5±5.6 years and 30 months respectively. Baseline mean sperm concentration was 29 million/ml, mean progressive motility 24% and a percentage DFI ranged from 5–36% with a mean of 16%. Sperm DNA integrity improved significantly with a DFI decrease from 25±11% to 18±6%, 3 months post-surgery which was consistent at 6 months (13±5%). Sperm concentration and progressive motility increased with a mean sperm count improvement by 21.9 million/ml and mean motility by 20.3%. Conclusions: Microscopic varicocelectomy provides durable improvement in DNA integrity and semen parameters. Sperm DNA integrity assessment using SCSA is a useful tool to demonstrate change in the semen quality post treatment.

The Correlation Between Sperm DNA Integrity and Conventional Semen Parameters

The integrity of sperm deoxyribonucleic acid (DNA) and chromatin is very important for genetic material transmission into offspring. Therefore, the aim of the present study was to investigate the correlation between sperm deoxyribonucleic acid integrity and sperm parameters. The current study was conducted on 96 semen samples. This study was done at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University for the period between November 2020 and May 2021. The samples were collected, and seminal fluid analysis was performed according to World Health Organization guidelines. Sperm Chromatin Dispersion test was performed. The results showed that the deoxyribonucleic acid fragmentation of sperms in infertile men in this study was higher than reported in fertile men in previous studies indicating that deoxyribonucleic acid fragmentation (DNA) is actively contributing at least partially to male infertility. In terms of enrolled men, sperm deoxyribonucleic acid fragmentation (SDF) showed significant positive correlation with round cell count.

The Correlation Between Sperm DNA Integrity and Conventional Semen Parameters

The integrity of sperm deoxyribonucleic acid (DNA) and chromatin is very important for genetic material transmission into offspring. Therefore, the aim of the present study was to investigate the correlation between sperm deoxyribonucleic acid integrity and sperm parameters. The current study was conducted on 96 semen samples. This study was done at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University for the period between November 2020 and May 2021. The samples were collected, and seminal fluid analysis was performed according to World Health Organization guidelines. Sperm Chromatin Dispersion test was performed. The results showed that the deoxyribonucleic acid fragmentation of sperms in infertile men in this study was higher than reported in fertile men in previous studies indicating that deoxyribonucleic acid fragmentation (DNA) is actively contributing at least partially to male infertility. In terms of enrolled men, sperm deoxyribonucleic acid fragmentation (SDF) showed significant positive correlation with round cell count

Detection of Sperm DNA Integrity and Some Immunological Aspects in Infertile Males

Immunoinfertility caused by anti-sperm antibodies (ASAs) represents about 10–20% of infertility among couples, which interfere with sperm motility and ability to penetrate cervical mucus, sperm-oocyte binding, fertilization, and embryo development. In addition, deoxyribonucleic acid (DNA) damages are increasingly found with infertile cases affecting male reproduction potency and progeny. This study aims to assess the semen, presence of ASAs, and DNA fragmentation index in normozoospermic patients. A total number of 116 cases with an average age of 20–51 years old, and duration of infertility at 4.70 ± 2.77 are classified into 77 and 39 primary and secondary types of infertility, respectively. Mixed agglutination reaction test was used to estimate the ASAs in semen (direct method) and in seminal plasma and blood serum (indirect method), for both immunoglobulins IgG and IgA. Acridine orange test was used to detect DNA fragmentation index. The results showed a significant difference (P > 0.05) for those with a secondary type of infertility at means 24.37 and 31.48 for IgG, and 14.46 ± 1.76 and 6.86 ± 0.39 for IgA by both direct and indirect methods, respectively. The direct method showed a significant difference only for the sperm tail, while that for indirect method was in sperm mid-piece. The mean of DFI for all cases was 38.25 ± 2.08, at 41.61 ± 2.19 and 31.63 ± 4.29, for both primary and secondary cases, respectively. The percentage of ASAs revealed no significant difference with DFI, except in some parts of sperm.

P-074 Sperm deoxyribonucleic acid integrity decreases with age and exhibits a rapid decline beyond the age of 35: a retrospective evaluation of 3446 semen samples

Abstract Study question Does advancing paternal age correlate with sperm DNA fragmentation index (DFI) and is there a cut-off age beyond which sperm DFI increases significantly? Summary answer In infertile men, DFI correlate with advancing paternal age and should be routinely screened starting 35 years of age. What is known already In recent decades, birth rates have substantially increased for men older than 30 years because of advanced age of marriage, rising life expectancy at birth, modern societal norms, and accessibility to assisted reproductive technology (ART). Advanced paternal age has been associated with a decline in conventional semen parameters (volume, concentration, motility, DFI), as well as, reduced fertility, increased risk of miscarriage, structural chromosomal aberrations and complex epigenetic disorders. Several studies have recommended testing sperm DFI in infertile men with advanced age (≥40 years) as it may provide prognostic information for couple attempting natural and assisted reproduction. Study design, size, duration This is a retrospective study of 3446 semen samples from patients under investigation for infertility between April 2016 and January 2022. Semen samples were obtained after 2-3 days of sexual abstinence. Patients were stratified into seven groups based on their age: patients ≤ 29 years (n = 127; 3.7%), 30-35 years (n = 868; 25.2%), 36-39 years (n = 863; 25.0%), 40-45 years (n = 1017, 29.5%), 46-49 years (n = 321; 9.3%), 50-55 years (n = 179, 5.2%) and ≥ 56 years (n = 71, 2.1%). Participants/materials, setting, methods Conventional semen parameters were assessed according to the WHO criteria and DFI was evaluated by TUNEL assay using the APODirect Kit run on BDAccuriC6 flow cytometer. Pearson’s r was used for correlation analysis between sperm concentration, DFI and paternal age. DFI results for each stratified patient group were evaluated by one-way ANOVA, followed by Tukey pos-hoc multiple comparison test. Results are presented as the mean±standard error and a P-value of < 0.05 was considered statistically significant. Main results and the role of chance In this cohort of men with a mean age of 39.5 years ± 0.1 (range 23-76 years), sperm deoxyribonucleic acid (DNA) fragmentation (21.1% ± 0.2) was positively correlated with age (r = 0.23, p˂0.001). In contrast, the correlation between sperm concentration and age was non-significant (r = 0.03, p = 0.07). Mean DFI in patients segregated into seven age groups were: ≤29 years (15.7% ± 0.8), 30-35 years (17.7% ± 0.4), 36-39 years (19.7% ± 0.4), 40-45 years (22.6% ± 0.4), 46-49 years (26.2% ± 0.9), 50-55 years (26.7% ± 1.2) and ≥ 56 years (31.1% ± 2.0). Mean %DFI level in the 26-29 and 30-35 age groups were non-significantly different (p = 0.65). However, mean %DFI level in the 36-39 age group was significantly higher than in the 26-29 and 30-35 age groups (p = 0.02 and p = 0.03, respectively). Mean %DFI level in the older age groups (40-45, 46-49, 50-55 and ≥ 56 years) were all significantly higher than in the 26-29 or 30-35 age groups (p˂0.001). Using a %DFI threshold level of 16.9%, 46.0% of patients ˂36, 52.2% of men aged 36-39, 60.2% of men aged 40-45, 67.3% of men aged 46-49, 72.6% of men aged 50-55 and 74.7% of men aged ≥56 years had an elevated DFI. Limitations, reasons for caution This is a retrospective analysis that did not account for confounding variables (e.g., clinical diagnosis, gonadotoxin exposure, febrile illness) that may affect conventional sperm parameters and DFI. Wider implications of the findings Our results underline the relationship between paternal age and sperm DFI and demonstrate a significant decline in sperm DNA fragmentation in men over the age of 35 years. The data suggest that we may want to reconsider the age cut-off we traditionally use to define advanced paternal age. Trial registration number No trial registration

The effect of swim-up and gradient sperm preparation techniques on deoxyribonucleic acid (DNA) fragmentation in subfertile patients.

To compare the effect of two different sperm preparation techniques, including swim-up and gradient methods on sperm deoxyribonucleic acid (DNA) fragmentation status of semen samples from unexplained and mild male factor subfertile patients undergoing intrauterine insemination (IUI). A prospective randomized study was conducted in 65 subfertile patients, including 34 unexplained and 31 male factor infertility to compare basal and post-procedure DNA fragmentation rates in swim-up and gradient techniques. Sperm DNA fragmentation rates were evaluated by a sperm chromatin dispersion (SCD) test in two portions of each sample of semen that was prepared with either swim-up or gradient techniques. Sperm motility and morphology were also assessed based on WHO 2010 criteria. Swim-up but not gradient method yielded a statistically significant reduction in the DNA fragmented sperm rate after preparation as compared to basal rates, in the semen samples of both unexplained (41.85 ± 22.04 vs. 28.58 ± 21.93, p < 0.001 for swim-up; and 41.85 ± 22.04 vs. 38.79 ± 22.30, p = 0.160 for gradient) and mild male factor (46.61 ± 19.38 vs. 30.32 ± 18.20, p < 0.001 for swim-up and 46.61 ± 19.38 vs. 44.03 ± 20.87, p = 0.470 for gradient) subgroups. Swim-up method significantly reduces sperm DNA fragmentation rates and may have some prognostic value on intrauterine insemination in patients with decreased sperm DNA integrity.

Sperm quality and DNA integrity of coke oven workers exposed to polycyclic aromatic hydrocarbons.

The objective of this study was to assess sperm quality and deoxyribonucleic acid (DNA) integrity of coke oven workers exposed to polycyclic aromatic hydrocarbons (PAHs) as compared to control subjects. The coke oven workers (N = 52) and administrative staff (N = 35) of a steel plant served as the exposed and control groups, respectively. Exposure to PAHs was assessed by measuring 1-hydroxypyren. Analysis of sperm quality (concentration, motility, vitality, and morphology) was performed simultaneously with sperm DNA integrity analysis, including DNA fragmentation, denaturation, bulky DNA adducts, and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dGuo). A questionnaire was conducted to collect demographic and potential confounding data. The coke oven workers had lower percentages of sperm motility, vitality and normal morphology than the control group, but the difference was not significant. For DNA integrity, the coke oven workers had significantly higher concentrations of bulky DNA adducts and 8-oxo-dGuo than the control subjects (p = 0.009 and p = 0.048, respectively). However, DNA fragmentation percentages did not significantly increase as compared to those in the subjects from the control group (p = 0.232). There was no correlation between sperm quality parameters and DNA integrity indicators. Occupational exposure of the coke oven workers to PAHs was associated with decreased sperm DNA integrity. Int J Occup Med Environ Health 2016;29(6):915-926.

Sperm deoxyribonucleic acid (DNA) damage assessment in male partners of couples with recurrent pregnancy loss and unexplained infertility: a prospective cohort study

Sperm DNA integrity plays a role in embryo development and fetal well being and may adversely affect the outcome of both natural and assisted conception in patients with unexplained recurrent pregnancy loss (RPL) and unexplained infertility. Our goals are:1-To determine if sperm DNA integrity plays a role in idiopathic recurrent pregnancy loss (RPL) and unexplained infertility in male partners with normal semen parameters. 2- To determine if the method of sperm processing affects the amount of sperm DNA fragmentation.

Impact of chemotherapeutics and advanced testicular cancer or Hodgkin lymphoma on sperm deoxyribonucleic acid integrity

To determine the impact of combination chemotherapy on the quality of newly generated spermatozoa from patients with advanced testicular cancer and patients with Hodgkin lymphoma (HL). Prospective longitudinal study. Academic facility. Patients with newly diagnosed metastatic testicular cancer and with HL that required chemotherapy were compared with age-matched healthy community volunteers. None. Semen parameters, hormone levels, testis volume, and presence of sperm DNA strand breaks in patients with cancer and in healthy community volunteers were compared before and after the patients' chemotherapy at 6, 12, 18, and 24 months. Before chemotherapy, both cancer groups had poorer semen quality compared with community volunteers. Among patients with testicular cancer and HL, 67% and 60%, respectively, had <5x10(6) sperm/mL at 6 months after chemotherapy. At 24 months, 60% and 57% of patients with testicular cancer and HL, respectively, had normal sperm concentrations. Level of FSH was significantly higher in the cancer group compared with community volunteers at 6 to 12 months after chemotherapy. Before chemotherapy, sperm DNA damage was higher in the cancer group than in community volunteers; this damage was increased further at 6 months and remained elevated 24 months after treatment. Sperm generated after chemotherapy maintain a significant degree of chromatin damage. Therefore, survivors of testicular cancer and HL are at risk of having abnormal reproductive outcome. Proper counseling to these patients on reproductive risks and fertility preservation before chemotherapy is recommended.

Effects of cryopreservation on human sperm deoxyribonucleic acid integrity

To evaluate the effect of cryopreservation on sperm motility and viability and to assess sperm DNA fragmentation and oxidation in men undergoing infertility investigation before and after cryopreservation in liquid nitrogen. Analysis of cryopreservation effects on sperm DNA integrity. Laboratory of Histology-Embryology of medicine faculty, Sfax, Tunisia. Fifteen semen samples from men undergoing infertility investigation. Neat semen samples were cryopreserved in liquid nitrogen using a commercial freezing medium (SpermFreeze, Fertipro, Belgium) according to the manufacturer's instructions. Samples were thawed at room temperature. Sperm DNA fragmentation was assessed using terminal deoxynucleotidyl transferase (Tdt) mediated dUTP nick end labeling and sperm DNA oxidation was determined using a fluorescent assay (OxyDNA test) for the detection of 8-oxoguanine. Evaluation of DNA fragmentation and oxidation rates was carried out before and after cryopreservation using flow cytometry. A significant decrease in sperm motility and viability was observed after cryostorage. In addition, sperm DNA fragmentation and DNA oxidative damage increased significantly after cryopreservation/thaw. Cryopreservation has deleterious effects on sperm DNA by inducing DNA fragmentation and oxidation but the mechanisms underlying such damages need to be elucidated by further investigations.

Intracytoplasmic sperm injection (ICSI) is a better choice than conventional in vitro fertilization (IVF) in assisted reproduction cycles with elevated sperm DNA fragmentation

Many studies have shown a strong correlation between sperm deoxyribonucleic acid (DNA) fragmentation and cycle outcome in couples undergoing assisted reproduction technologies (ART). However, there is no agreement on whether IVF or ICSI provide a better chance for the couples when sperm DNA fragmentation is abnormally high (20%), although the other seminal parameters are normal. The aim of this study is to evaluate the sperm DNA integrity by comparing the rate of sperm DNA fragmentation in semen samples from infertile men and their fertilizing capacity and implantation rates in both conventional IVF and ICSI procedures. Prospective study of 45 couples from our laboratory IVF/ICSI program (from January 2006 to October 2006). We analyzed 45 couples that underwent a combined IVF/ICSI cycle after 4 failed intrauterine inseminations (IUI). Half of the retrieved oocytes from each couple were fertilized by ICSI and the other half by conventional IVF. After density gradient, sperm DNA fragmentation was measured by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-nick end labelling assay (TUNEL) with flow cytometry. Sperm DNA fragmentation was considered abnormal if values were >20%. Out of the 45 couples evaluated, 13 (29%) had an abnormal sperm DNA fragmentation test results. Fertilization rate with IVF was significantly reduced when compared with ICSI in these patients showing a high sperm DNA fragmentation (51% vs. 74%, P=0.027). Similarly, mean number of good quality embryos available for transfer in couples with high sperm DNA fragmentation was higher after ICSI (1.45 ± 0.7 vs. 0.85 ± 0.6, P=0.043). No significant differences were found in both implantation and pregnancy rates. Our results suggest that ICSI may be a better option than conventional IVF for those couples undergoing ART and showing an abnormally high sperm DNA fragmentation.

High risk of temporary alteration of semen parameters after recent acute febrile illness

To report parameters in semen samples and sperm deoxyribonucleic acid integrity in a fertile volunteer presenting a 2-day fever of 39 degrees -40 degrees C. Case report. University-affiliated teaching hospital. None. Semen samples from a fertile volunteer of proven fertility were obtained and analyzed before the febrile illness episode and at days 15, 37, 58, 79, and >180 after the fever. Semen parameters (total sperm count, motility a+b, and vitality), sperm protamination state, measured by sperm chromatin structure assay (SCSA) and apoptotic activities, measured by terminal uridine nick-end labeling (TUNEL) assay. Total sperm count significantly decreased at days 15, 37, and 58 after the fever and returned to normal by day 79 after the fever. The percentage of motility significantly decreased at days 15 and 37 after the fever and returned to normal by day 58. Vitality score also showed a slight, although not statistically significant, decrease after the fever. The DNA fragmentation index (DFI, a SCSA parameter), which defines abnormal chromatin structure, significantly increased by 24% and 36% at days 15 and 37 after the fever, respectively, and decreased to 15% and 8% when reaching days 58 and 79 after the fever. High DNA stainability (HDS, a SCSA parameter) also significantly increased at day 37 after the fever. On the other hand, sperm DNA fragmentation, as measured by TUNEL assay, increased up to 23% by day 15 after the fever but this was not statistically significant. This report demonstrates that a febrile episode can have marked effects on semen parameters and sperm DNA integrity. These results are particularly important for the counseling of infertile couples and in relation to assisted reproductive techniques (ART).

Leukocytospermia: relationship to sperm deoxyribonucleic acid integrity in patients evaluated for male factor infertility

The purpose of our study was to analyze the relationship of leukocytospermia to sperm DNA damage as assessed by the DNA integrity assay. Results indicate that leukocytospermia has a significant negative effect on the standard semen parameters of concentration, motility, and morphology but at most a very weak effect on DNA integrity as measured by the DNA fragmentation assay.

Ten-year variation in semen parameters and sperm deoxyribonucleic acid integrity in a healthy fertile man

To report parameters in semen samples and sperm DNA integrity in a healthy fertile volunteer over a 10-year period. Case report. University-affiliated teaching hospital. None. Semen samples from a nonsmoking healthy male volunteer of proven fertility aged from 40 to 50 years were collected and analyzed over a decade. Semen parameters (sperm count, total sperm count, percentage of progressive motility grades a+b, morphology, and percentage of living spermatozoa) and sperm DNA integrity, measured by sperm chromatin structure assay (SCSA) and terminal uridine nick-end labeling (TUNEL) assay. Median (min-max) value of total sperm count was 330 (126-511) million. Motility and vitality presented a median of 50% (40%-75%) and 78% (53%-92%), respectively. Among semen parameters, morphology and vitality showed the lowest within-subject coefficient of variation (CV(W)) and the total sperm count the highest (8.1% and 12.0% vs. 34.9%). Median values of DNA fragmentation index (DFI) and high DNA stainability (HDS) were 12.7% (7.9%-16.5%) and 6.5% (5.5%-8.2%), respectively. Sperm DNA fragmentation presented a median value of 8.9%, a minimum value of 1.4% and maximum value of 18.6%. Compared with TUNEL data, SCSA parameters (DFI and HDS) showed less variation over the data collection period (47.4% vs. 22.4% and 13.0%, respectively). Our data show that in this healthy fertile volunteer, semen parameters and sperm DNA integrity remained normal, and no trend was observed over the study period. More interestingly, in this subject aged from 40 to 50 years old, sperm nucleus status presented less than 20% of sperm DNA fragmentation over a decade.

Age-related decline in sperm deoxyribonucleic acid integrity in patients evaluated for male infertility

The purpose of this study was to analyze the relationship of male age and DNA integrity in men presenting for investigation of infertility. We found that men 45 years and older had significantly greater DNA fragmentation than younger men.

Human sperm deoxyribonucleic acid fragmentation by specific types of papillomavirus

Objective: Human papillomaviruses are present in up to 64.3% of patients’ sperm. The objectives were (1) to determine human papillomavirus deoxyribonucleic acid effects on sperm deoxyribonucleic acid integrity and (2) to assess human papillomavirus differential effects on the sperm cell. Study Design: Two-layer colloid washed sperm were exposed to E6-E7 deoxyribonucleic acid fragments generated from human papillomavirus types 16, 18, 31, 33, 6/11, or control (DQA1) for 24 hours. The motility parameters were measured and analyzed. Pilot studies were performed to develop a fixed sperm comet assay to assess deoxyribonucleic acid fragmentation. Results: Significant sperm deoxyribonucleic acid fragmentation occurred after exposure to deoxyribonucleic acid of human papillomavirus types 16 and 31. Human papillomavirus deoxyribonucleic acid fragment size was not a factor. Human papillomavirus types 18, 33, and 6/11 did not compromise sperm deoxyribonucleic acid integrity. Washed sperm motility was higher in the presence of human papillomavirus deoxyribonucleic acid except for type 6/11. Amplitude of head displacement was lower for human papillomavirus types 16 and 6/11. Sperm linearity was increased for all human papillomavirus types except type 18. Conclusion: Human papillomavirus type 16 and 31 deoxyribonucleic acid caused deoxyribonucleic acid breakages characteristic of apoptotic but not necrotic sperm. The data suggest that these human papillomavirus types may adversely affect subsequent embryonic development after fertilization. Sperm deoxyribonucleic acid appears to resist human papillomavirus types 18, 33, and 6/11 or repairing mechanisms occurred. Although enhanced motility was found in human papillomavirus–exposed sperm, important velocity parameters were decreased, suggesting impaired sperm function. (Am J Obstet Gynecol 2001;184:1068-70.)

Potential adverse effect of semen processing on human sperm deoxyribonucleic acid integrity

Objective: To examine the effect of standard Percoll density-gradient centrifugation on human sperm DNA denaturation. Design: Prospective, observational study. Setting: University-based infertility clinic. Patient(s): Twenty-five nonazoospermic men. Intervention(s): Semen samples (n = 25) were obtained from consecutively seen nonazoospermic men presenting for infertility evaluation. Samples were processed by two-layer and four-layer Percoll density gradients. Sperm motility and sperm chromatin structure (evaluated by flow cytometry analysis of acridine orange–treated spermatozoa) were monitored before and after semen processing. Sperm chromatin integrity was expressed as the percentage of spermatozoa that demonstrated denatured DNA. Main Outcome Measure(s): Sperm motility and DNA integrity. Result(s): Mean sperm motility improved significantly after processing with two-layer and four-layer Percoll gradients compared with whole semen (54% and 57% motility versus 44% motility, respectively). In contrast, the percentage of sperm with denatured DNA increased after processing with two-layer and four-layer Percoll gradients compared with whole semen (34% and 32% versus 18%, respectively). Conclusion(s): Our data demonstrate that the improvement seen in sperm motility after Percoll processing is not associated with a similar improvement in sperm DNA integrity. These data suggest that we reexamine current sperm processing techniques to minimize sperm DNA damage and the potential transmission of genetic mutations in assisted reproductive cycles.