Impact Of Sperm DNA Fragmentation Research Articles

Impact of sperm DNA fragmentation in couples with unexplained recurrent pregnancy loss: A cross-sectional study.

Recurrent pregnancy loss (RPL) has a multifactorial etiology, with a majority of cases remaining unexplained. To account for these unexplained cases, possible male factors are being explored. Conventional semen analysis lacks a qualitative assessment of sperms and information regarding sperm DNA integrity. Sperm DNA fragmentation (SDF) has diagnostic value in unexplained RPL, and it may account for a number of unexplained cases. Hence, we planned a study to explore and evaluate the impact of sperm DNA fragmentation in couples with unexplained recurrent pregnancy losses. Analytical cross-sectional study was conducted at a tertiary-level referral facility in India between August 2021 and July 2023. Participants (n = 70) were divided into two groups-male partners of couples with unexplained RPL (following spontaneous conceptions) (n = 35) and men with at least one previous live birth (spontaneous or following fertility treatments for female factor infertility such as ovulation induction or intrauterine insemination) as controls (n = 35). Neither of the two groups of couples recruited for this study had undergone ART as fertility treatment. Primary outcome assessed was mean DNA fragmentation index (DFI). Secondary outcomes included differences in semen parameters such as sperm concentration, progressive sperm motility and morphology, proportion of men with high (≥30%) and low DFI in the two groups, and the association between various semen parameters and DFI. Univariate logistic regression revealed that sperm DNA fragmentation was higher in men with unexplained RPL (30.0; IQR (interquartile range) 19.0, 46.0) as compared to controls (22.0; IQR 14.0, 30.0) although it was not statistically significant (OR, odds ratio, 1.02; 95% CI 1.0-1.1, p = 0.08). A higher proportion of men with unexplained RPL had DFI ≥30% compared to controls (54.2% vs. 25.7%; OR 3.43 (95% CI 1.2-9.4); p = 0.02). No statistically significant differences were observed in semen volume, sperm concentration, progressive motility, and morphology between the two groups. Sperm DNA fragmentation index also showed a weak but significant inverse relationship with sperm morphology (r = -0.336, p = 0.004). The current study did not show any significant difference in the mean sperm DNA fragmentation levels in male partners of couples with unexplained RPL compared to controls. However, a higher proportion of men with DFI ≥30% were observed in unexplained RPL population when compared to controls.

Elevated sperm DNA fragmentation is correlated with an increased chromosomal aneuploidy rate of miscarried conceptus in women of advanced age undergoing fresh embryo transfer cycle.

Male sperm DNA fragmentation (SDF) may be associated with assisted reproductive technology (ART) outcomes, but the impact of SDF on the occurrence of aneuploid-related miscarriage remains controversial. Genome-wide single-nucleotide polymorphism-based chromosomal microarray analysis was performed on 495 miscarried chorionic villus samples undergone IVF/ICSI treatment from the Reproductive Medicine Center of the First Affiliated Hospital of Zhengzhou University. SDF was assessed using sperm chromatin structure assay. Patients were divided into four groups according to embryo transfer cycle type and maternal age, and the correlation between SDF and chromosome aberration was analyzed. A receiver operating characteristic (ROC) curve was utilized to find the optimal threshold. Total chromosomal aneuploidy rate was 54.95%, and trisomy was the most common abnormality (71.32%). The chromosomally abnormal group had higher SDF than the normal group (11.42% [6.82%, 16.54%] vs. 12.95% [9.61%, 20.58%], P = 0.032). After grouping, elevated SDF was significantly correlated with an increasing chromosome aneuploidy rate only in women of advanced age who underwent fresh embryo transfer (adjusted odds ratio:1.14 [1.00-1.29], adjusted-P = 0.045). The receiver operating characteristic curve showed that SDF can predict the occurrence of chromosomal abnormality of miscarried conceptus in this group ((area under the curve = 0.76 [0.60-0.91], P = 0.005), and 8.5% was the optimum threshold. When SDF was ≥ 8.5%, the risk of such patients increased by 5.76 times (adjusted odds ratio: 6.76 [1.20-37.99], adjusted-P = 0.030). For women of advanced maternal age undergoing fresh embryo transfer, older oocytes fertilized using sperm with high SDF in IVF/ICSI treatment might increase the risk of chromosomal abnormality in miscarried conceptus.

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.

O-251 Defining the exclusive role of male genome integrity on conceptus development

Abstract Study question To determine the effect of sperm DNA fragmentation on embryo development by ruling out a female factor component. Summary answer By utilizing healthy donor oocytes, it is possible to quantify the sole deleterious effect of sperm DNA fragmentation and explore the ooplasmic repair mechanism. What is known already In approximately 50% of couples with unexplained ART failure, a subtle male factor is present that is missed in a conventional semen analysis assessing concentration, motility, and morphology. Additional information on male gamete competence can be acquired by assessing Sperm Chromatin Fragmentation (SCF). Indeed, a fragmented male genome can give rise to poorly developing embryos, leading to impaired implantation, lower pregnancy, and higher miscarriage rates. It has been previously seen that an oocyte, according to its age, can repair the damaged DNA contributed by the male gamete. Study design, size, duration In last decade, 316 couples, who have an elevated SCF, underwent ICSI cycles and resulted in disappointing clinical outcomes. To exclude an eventual confounding female factor, couples who utilized donor oocytes were identified and clinical outcome was compared to a control. To measure the role of an impaired sperm genome and a concurrent oocyte repair mechanism, we compared clinical outcomes between cycles in the same couples who used their own and subsequently donor oocytes. Participants/materials, setting, methods We included 381 couples screened for SCF; 65 underwent ICSI cycles with donor oocytes; of these, 46 underwent a previous ICSI cycle with their own oocytes. Fertilization, implantation, clinical pregnancy, and delivery rates were compared between above-mentioned groups. A TUNEL assay was used to measure SCF (≥500 spermatozoa were assessed/sample at a 15% threshold). Paired t and Chi-square tests were used to compare ages and clinical outcomes, respectively. Main results and the role of chance We included 381 couples (maternal age, 37.8 ± 4.2; paternal age, 41.8 ± 8.2), whose male partners had the following semen parameters: average volume of 2.5 mL, concentration of 25.4x106/mL, 33% motility, and normal morphology of 1.7. Of these, 316 had elevated average DNA fragmentation of 25.5% and utilized their own oocytes in 683 ICSI cycles, resulting in a 67.5% fertilization, 12.2% implantation, 27.9% clinical pregnancy, and a 22.5% ongoing/delivery. Of the 381 couples, 65 underwent ICSI utilizing donor oocytes (SCF of 24.3%) resulting in a significantly higher fertilization (78.4%) (P < 0.00001) and embryo implantation rates (34.2%) (P < 0.0001). Similarly, albeit not statistically significant, the clinical pregnancy and ongoing/delivery rates increased from 27.9% to 37.1% and from 22.5% to 30.0%, respectively. To further explore the exclusive role of the male gamete in embryo development, we identified a cohort of patients (n = 46), with an elevated average DNA fragmentation of 23.6%, who underwent an ICSI cycle using their own oocytes and a subsequent cycle using donor oocytes. Compared to cycles where couples used their own oocytes, fertilization, embryo implantation, clinical pregnancy, and ongoing/delivery rates rose from 67.5% to 76.6% (P < 0.0001), 2.8% to 24.7% (P < 0.00001), 8.4% to 38.8% (P < 0.0001), and 1.2% to 28.6% (P < 0.00001), respectively. Limitations, reasons for caution Although this study attempts to control for a concurrent confounding female factor, it cannot be completely excluded. It is difficult to assess the extent and contribution of the ooplasmic repair mechanisms on the male genome. Wider implications of the findings Ooplasmic repair mechanisms of healthy female gametes appear to repair or improve the deleterious impact of sperm DNA fragmentation on ART outcomes. Trial registration number not applicable

P-099 The effect of sperm DNA quality on blastocyst and euploid rates

Abstract Study question Do sperm DNA quality measurements influence the blastocyst rate and/or euploid rates? Summary answer Sperm DNA fragmentation has a negative influence on the rate of blastulation. It does not impact on euploidy rates assessed by pre-implantation genetic testing (PGT-A). What is known already Some studies show that sperm DNA has no influence on embryo development and that its negative influence begins following implantation. Other studies show that every step of the fertility process from fertilization onwards is impacted by the male gamete as well as the female gamete. To date, data has been confounded by different sperm DNA tests and different clinical practices and parameters measured. Study design, size, duration This was a retrospective study using the sensitive Comet assay to analyse whether it’s three sperm DNA fragmentation scores influence embryo development or euploidy. Participants/materials, setting, methods Data from 110 cycles of intracytoplasmic sperm injection (ICSI) where the blastocyst rate (number of blastocysts/number of mature oocytes injected) was obtained from the patient database of a UK single centre. Inclusion criteria: Only outcomes from the first assisted reproductive cycle after a Comet assay had been performed were included. Included cycles were within one year of the test. Main results and the role of chance Blastocysts were ranked in 4 groups- 0.25-0.49, 0.5-0.74, 0.75-1.00. Three Comet scores were analysed: Average Comet Score (damage across the semen sample; ACS), Low Comet Score (proportion of sperm with good DNA; LCS) and High Comet Score ( proportion of sperm with high DNA damage; HCS). Clinical thresholds were calculated by comparison with fertile donor sperm. High ACS and LCS and low HCS were all associated with higher blastocyst rates (ANOVA, p = 0.013; Kruskal Wallis, p = 0.027, Post Hoc pairwise comparisons between blastocyst groups (p = 0.016-0.024). No relationship was found between euploid rate (number of euploid embryos/total embryo number) and sperm DNA damage p = 0.761). Limitations, reasons for caution Retrospective design requiring more data to clarify these associations coupled with the lack of randomisation and low power analysis. Wider implications of the findings This study reports the negative impact of sperm DNA fragmentation on blastocyst formation rates. Male partner diagnostic assessments such as sperm DNA quality testing are useful adjuncts to semen analysis. Further research to determine if there is a relationship between sperm DNA damage and euploid embryo outcomes is ongoing. Trial registration number IRB-001C06-01-22

Impact of sperm DNA fragmentation on the clinical outcome of assisted reproduction techniques: a systematic review of the last five years.

To elucidate through a systematic literature review the impact sperm DNA fragmentation has on embryos from assisted reproduction techniques. Studies from the "PubMed", "Embase", and "BVS" databases were analyzed. The articles selected in the review included: cohort and case-control studies that addressed the proposed theme, published between January 1, 2017, and January 31, 2022, in English, Portuguese, and Spanish. As inclusion criteria: cohort and case-control articles. As exclusion criteria: articles outside the scope of the research, review articles, case reports, articles using animal models, abstracts, letters to the editor, and articles found duplicates in the databases. Number of couples or cycles; age (men/women); collection type; DNA damage (%); assisted reproduction activity and techniques. In in vitro fertilization, a reduction in fertilization rate, blastocyst rate, and embryo quality was observed. In addition to implantation and increased abortion rates in patients with high sperm DNA fragmentation. High rates of sperm DNA fragmentation in intracytoplasmic sperm injection led to reduced blastocyst production rate, embryo quality, implantation, and live birth rate, and in intrauterine insemination, a reduction in pregnancy rate. Sperm DNA fragmentation was a potential limiting factor for assisted reproduction techniques.

Beyond conventional sperm parameters: the role of sperm DNA fragmentation in male infertility.

Infertility is a condition that widely affects couples all over the world. In this regard, sperm DNA fragmentation can lead to harmful reproductive consequences, including male infertility and poor outcomes after assisted reproductive techniques. The investigation of sperm DNA fragmentation in male infertility diagnostics has constantly increased over time, becoming more common in clinical practice with the recent publication of several guidelines regarding its testing. This narrative review aims to provide a comprehensive overview of the pathogenesis and causes of sperm DNA fragmentation, as well as the assays which are commonly performed for testing. Moreover, we discuss the most recently published evidence regarding the use of sperm DNA fragmentation testing in clinical practice, highlighting the implications of high sperm DNA fragmentation rate on human reproduction, and the therapeutic approaches for the clinical management of infertile patients. Our review confirms a significant harmful impact of sperm DNA fragmentation on reproduction and points out several interventions which can be applied in clinics to reduce sperm DNA fragmentation and improve reproductive outcomes. Sperm DNA fragmentation has been shown to adversely impact male fertility potential. As high sperm DNA fragmentation levels have been associated with poor reproductive outcomes, its testing may significantly help clinicians in defining the best therapeutic strategy for infertile patients.

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 (<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 <30% SDF had the highest fertilization rate (p = 0.05, p < 0.01 and p < 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 < 0.01), smooth endoplasmic reticulum clusters (p < 0.01), large perivitelline space (p < 0.01), resistant membrane (p < 0.01), and non-resistant membrane (p < 0.01), while the influence of SDF index on the pregnancy rate was influenced by the presence large perivitelline space (p < 0.01), resistant membrane (p = 0.018) and non-resistant membrane (p < 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

P–058 The effect of sperm DNA fragmentation on intracytoplasmic sperm injection (ICSI) outcome

Abstract Study question Does the sperm DNA fragmentation (SDF) level impact the clinical outcome of couples undergoing intracytoplasmic sperm injection (ICSI)? Summary answer No significant effect was observed for SDF on the reproductive outcome of couples undergoing ICSI. What is known already Sperm DNA Fragmentation (SDF) has emerged as an important biomarker in the assessment of male fertility potential. It is currently being used as one of the advanced sperm function tests along with other conventional methods in male fertility evaluation. The impact of SDF on the reproductive outcomes of ICSI remains to be controversial. Evidence extracted from three meta-analyses have indicated that higher SDF is not associated with a negative impact on ICSI outcomes. On the contrary, another meta-analysis revealed that SDF can have a significant impact on the pregnancy rate of ICSI with an OR of 1.31. Study design, size, duration This is a retrospective cohort study carried out in the assisted conception unit of a tertiary medical center. The study duration was over a 5-year period from August 1st, 2014 to August 1st, 2019. The charts of 1922 patients who underwent ICSI were screened for inclusion in the study. Inclusion criteria were patients who underwent ICSI using ejaculate spermatozoa and had a recorded SDF test done within a week before ICSI (n = 390). Participants/materials, setting, methods Sperm chromatin dispersion was used to evaluate SDF utilizing the Halosperm G2 test kit (Halotech, Madrid, Spain). All patients performed the ICSI trial using ejaculated spermatozoa. Patients were divided according to the SDF level into 3 groups; SDF <20% (n = 148), SDF 20–30% (n = 133), and SDF >30% (n = 109). Female partner fertility status was recorded and couples were grouped into 2 groups based on age and AMH levels; (1) favorable female and (2) unfavorable female status. Main results and the role of chance Overall, clinical pregnancy occurred in 45% of cases, live birth rate was 33.60%, and 1.30% of patients had miscarriage. A significant negative correlation between SDF and sperm count (r–0.232), motility (r–0.469), progressive motility (r–0.312) and normal morphology (r–0.297) was detected (p < 0.001 for all). Fertilization rate, clinical pregnancy and live birth rate were greater in patients with lower SDF than those with higher SDF in both favorable and unfavorable groups, however the difference was not statistically significant (Table 1). Limitations, reasons for caution The main limitation of our study was the retrospective nature of the study where some data may be missing or incomplete. The data was also retrieved from one ART center, therefore our data lacked diversity within methodologies for IVF and SDF testing. Wider implications of the findings: SDF was found to be significantly correlated with conventional semen parameters highlighting its significance as a robust diagnostic test during male fertility evaluation. In this study, while patients with higher SDF values had worse reproductive outcomes with ICSI, the results did not reach statistical significance. Trial registration number NA

Oocyte ability to repair sperm DNA fragmentation: the impact of maternal age on intracytoplasmic sperm injection outcomes

To study the impact of sperm DNA fragmentation (SDF) on clinical outcomes of assisted reproductive technology in women with different age ranges. Historical cohort study. Private university-affiliated invitro fertilization center. Five hundred forty couples undergoing intracytoplasmic sperm injection cycles. Cycles were split into three groups according to maternal age: ≤36 years old (n = 285), 37-40 years old (n = 147), and >40 years old (n = 108). Semen samples were evaluated for SDF using the Sperm Chromatin Dispersion test and, for each age group, the cycles were subdivided according to SDF index: low fragmentation index (<30% SDF) and high fragmentation index (≥30% SDF). Implantation, pregnancy, and miscarriage rates. For younger patients (≤36 years old) and those between 37 and 40 years of age, no significant differences were noted in laboratory and clinical outcomes for cycles with <30% SDF or ≥30% SDF. When maternal age was >40 years of age, significantly lower high-quality day-3 embryos (54.4% vs. 33.1% and blastocyst development rates (49.6% vs. 30.2%), lower pregnancy (20.0% vs. 7.7%) and implantation rates (19.7% vs. 11.9%), and increased miscarriage rate (12.5% vs. 100.0%) were observed for cycles with ≥30% SDF compared with <30% SDF, respectively. Older oocytes, when injected with sperm derived from samples with high SDF index, develop into embryos of poor quality that lead consequently to lower implantation and pregnancy rates and higher miscarriage rates, in intracytoplasmic sperm injection cycles from women with advanced maternal age.

Sperm DNA fragmentation testing: Summary evidence and clinical practice recommendations.

We herein summarise the evidence concerning the impact of sperm DNA fragmentation in various clinical infertility scenarios and the advances on sperm DNA fragmentation tests. The collected evidence was used to formulate 41 recommendations. Of these, 13 recommendations concern technical aspects of sperm DNA fragmentation testing, including pre‐analytical information, clinical thresholds and interpretation of results. The remaining 28 recommendations relate to indications for sperm DNA fragmentation testing and clinical management. Clinical scenarios like varicocele, unexplained infertility, idiopathic infertility, recurrent pregnancy loss, intrauterine insemination, in vitro fertilisation/intracytoplasmic sperm injection, fertility counselling for men with infertility risk factors and sperm cryopreservation have been contemplated. The bulk evidence supporting the recommendations has increased in recent years, but it is still of moderate to low quality. This guideline provides clinicians with advice on best practices in sperm DNA fragmentation testing. Also, recommendations are provided on possible management strategies to overcome infertility related to sperm DNA fragmentation, based on the best available evidence. Lastly, we identified gaps in knowledge and opportunities for research and elaborated a list of recommendations to stimulate further investigation.

The impact of sperm DNA fragmentation on ICSI outcome in cases of donated oocytes.

The aim of this study is to evaluate the sperm DNA fragmentation index (DFI) in oocyte donation cycles and correlate it with the sperm parameters, the male characteristics, the embryo quality and the outcome of intracytoplasmic sperm injection (ICSI). A total of 150 couples participating in an oocyte donation program were included in the study. Sperm samples were assessed by conventional sperm analysis. DFI was evaluated using the Halosperm kit, a sperm chromatin dispersion test (SCD). The relations between DNA damage and epidemiological male factors (age, height, weight), standard semen parameters (concentration, total and forward motility, and morphology), and embryological and clinical parameters (fertilization rate, total blastocyst number, number of good quality blastocyst, clinical pregnancy) were analyzed. DFI was positively correlated with advanced male age (r = 0.23, p < 0.05) and negatively correlated with total sperm and forward motility (r = -0.29, r = -0.27, respectively; p < 0.05). DFI was not significantly correlated with pregnancy outcome in oocyte donation cycles (r = -0.05, p > 0.05). When good quality blastocysts were chosen, a trend toward the development of good quality embryos was detected in the presence of a low DFI (r = -0.20, p = 0.08). DFI does not significantly affect the outcome of ICSI in oocyte donation cycles. Even in cases of advanced paternal age that a high DFI resulted sperm DNA fragmentation seems not to adversely affect the final outcome.

Impact of sperm DNA fragmentation on ICSI outcome and incidence of apoptosis of human pre-implantation embryos obtained from in vitro matured MII oocytes

BackgroundSperm DNA integrity and oocyte quality significantly affect embryo development and survival. The current study evaluated embryo development and quality, as well as the expression level of apoptosis-related genes and microRNAs in embryo derived from in vitro matured MII oocytes according to sperm DNA fragmentation (SDF) level. MethodsThe semen and immature oocytes were collected from 50 ICSI cycles with any recognizable female factor infertility. After ovarian stimulation, germinal vesicle stage (GV) oocytes were collected and incubated in in vitro maturation (IVM) medium for 24 h. Next, reactive oxygen species (ROS) level of media culture was determined. Using by sperm chromatin dispersion (SCD) test, the SDF levels of processed semen were assessed and categorized into SDF ≤ 30% and SDF>30%. Seventy two hours after intracytoplasmic injection, the embryo development and quality score were recorded in the groups I (GV-MII + SDF≤ 30%) and II (GV-MII + SDF> 30%). Also, the apoptosis incidence of embryos at morula stage was evaluated at molecular and cellular levels by quantitative real time PCR and TUNEL staining, respectively. ResultsCleavage rate did not differ between two groups. The quality score of embryos obtained from IVM matured oocytes and high level of SDF was significantly lower than that of low level of SDF (P < 0.05). The embryos from group II had a significant reduction of the expression of BCL-2 compared to group I (P < 0.05). Also, they showed an increase in relative transcription of pro-apoptotic microRNAs; miR 15a and miR 16-1 versus group I (P < 0.05). A rise of TUNEL positive blastomers of embryo was observed at group II versus group I, but it did not reach to significantly level. ConclusionThe IVM oocytes, probably, did not suffice to recover the high level of paternal genomic damage and inhibition of apoptosis pathway beginning.

Sperm DNA Fragmentation: Consequences for Reproduction.

DNA fragmentation, or the accumulation of single- and double-strand DNA breaks, is a common property of sperm, and an increase in the level of sperm DNA fragmentation is known to influence natural reproduction. The effect of sperm DNA fragmentation on male infertility and assisted reproductive treatment (ART) outcomes remains controversial and is one of the most frequently debated topics of reproductive medicine. For the past 30years, a number of assays have been developed to quantify the level of sperm DNA fragmentation. In this chapter, we review the causes of sperm DNA fragmentation, describe the commonly used tests to evaluate these abnormalities, and perform a systematic review of existing studies to determine the impact of sperm DNA fragmentation on male fertility and ART outcomes.

Associations of sperm DNA fragmentation with lifestyle factors and semen parameters of Saudi men and its impact on ICSI outcome

BackgroundMale factor infertility is quite common as 30–50% of infertility cases are due to sperm defects. The high sperm DNA fragmentation is one of the causes of male factor infertility. Many factors cause sperm DNA fragmentation and could be testicular or post-testicular. The purpose of this study was to assess relationships among sperm DNA fragmentation, lifestyle factors and semen values of Saudi men and to determine impact of sperm DNA fragmentation on ICSI cycle outcome.MethodsThe duration of this study was from January 2015 to June 2016. The cases with female factor infertility were excluded. In total 94 couples were selected for investigation. The study parameters were male age, body mass index, smoking, semen values, % sperm DNA fragmentation, fertilization rate and pregnancy outcome. The ICSI procedure was performed in all patients per standard protocol. The semen samples were grouped based on % sperm DNA fragmentation into < 15%, 15–30 and > 30% which corresponded to low, moderate and high sperm DNA fragmentation, respectively.ResultsThere was no difference in ICSI outcome in low and moderate sperm DNA fragmentation, however, in high sperm DNA fragmentation no patient achieved pregnancy. In this study, 53.19% Saudi men had low, 32.98% moderate and 13.83% high DFI. Semen volume, sperm morphology and fertilization rate did not show any correlation trend with DNA fragmentation, however, sperm concentration and motility were negatively correlated in all DFI categories. The BMI was positively correlated in moderate DFI category and smoking was positively correlated with low DFI category. The age was positively correlated in moderate and high DFI categories.ConclusionsThe results of this study indicated that 14% Saudi men had high DNA fragmentation. The BMI was positively correlated in moderate DFI category and smoking was positively correlated with low DFI category. The age was positively correlated in moderate and high DFI categories.

Влияние фрагментации ДНК сперматозоидов на эффективность оплодотворения и развитие эмбрионов человека, культивируемых in vitro

Влияние фрагментации ДНК сперматозоидов на эффективность оплодотворения и развитие эмбрионов человека, культивируемых in vitro

Impact of sperm DNA fragmentation on clinical in vitro fertilization outcomes.

ObjectiveWe studied the association between sperm DNA fragmentation (SDF) and several clinical in vitro fertilization outcomes.MethodsWe retrospectively analyzed 169 consecutive fresh IVF cycles. Semen was collected on the day of oocyte retrieval, and we assessed standard semen parameters and the SDF level (by terminal deoxynucleotidyl transferase dUTP nick-end labeling). Poor ovarian response (POR) was defined as the collection of three or fewer mature oocytes. Oocytes were inseminated by the conventional method or intracytoplasmic sperm injection.ResultsSDF did not affect the fertilization or pregnancy rate, but did have a significant effect on the miscarriage rate. In the miscarriage group (n=10), the SDF level was significantly higher (23.9% vs. 14.1%) and number of mature oocytes was significantly lower (4.3 vs. 7.6) than in the live birth group (n=45). Multiple regression analysis showed that SDF was an independent predictor of miscarriage (odds ratio, 1.051; 95% confidence interval, 1.001–1.104). The cutoffs for the SDF level and number of mature oocytes that could predict miscarriage were >13% and ≤3, respectively. In the low-SDF group (≤13%), the miscarriage rate was similar in POR patients and those with a normal ovarian response (NOR; 14.2% vs. 4.3%). In the high-SDF group (>13%), the miscarriage rate was significantly higher in the POR group than in the NOR group (60.0% vs. 13.3%, p=0.045).ConclusionOur study demonstrated that a high SDF level (>13%) was associated with a high miscarriage rate, and that it mainly contributed to miscarriage in the POR group. The results suggest that SDF measurements should be considered in couples with POR in order to predict the prognosis of the pregnancy.

DECREASED FERTILIZATION: HUMAN SPERM DNA FRAGMENTATION AND IN VITRO MATURATION OF OOCYTE IN STIMULATED ICSI CYCLES.

The aim of this study was to evaluate the impact of sperm DNA fragmentation (SDF) on fertilization rate, and sperm nuclear decondensation after intracytoplasmic injection of sperm (ICSI) into cumulus-free germinal vesicle (GV) oocytes from stimulated cycles. After the retrieval of oocytes, the GV oocytes were cultured for 24 h. Oocytes that liberated polar bodies were injected with processed semen, and were used to evaluate SDF level. Based on SDF, the data were categorized into two groups. Group I in which SDF≤ 30% and group II in which SDF > 30%. Fertilization (presence of two pronuclei) was checked 16-19 h after ICSI. Unfertilized oocytes were stained by Hoechst 33258 and examined to evaluate the undecondensed sperm head in oocyte. The rates of maturation, fertilization in fertilized IVM oocytes and the percentage of undecondensed sperm in the unfertilized oocytes were assessed according to SDF. Out of 146 GV oocytes that were subjected to IVM, 101 (69 %) developed to metaphase II. The fertilization rate of IVM oocytes in group II was significantly lower than that in group I (P < 0.05). Moreover, group I, had 25 % of their unfertilized oocytes containing condensed sperm, while group II had a significantly higher number (53 %) of unfertilized oocytes containing condensed sperm (P < 0.05). SDF had a negative effect on the rate of fertilization in matured in vitro GV oocytes and could lead to an increase in the percentage of undecondensed sperm in IVM oocytes from stimulated cycles.

Alcohol-induced male infertility: Is sperm DNA fragmentation a causative?

Background: There is a passionate desire for couples to own their own biological children. Unfortunately, infertility index has been increasing with about 50% attributed to male factor infertility. Sperm DNA fragmentation (SDF) has been suggested as one of the causes of infertility in men; however, there have been controversies as regards its relationship with the successful management of infertility. Aim: This study is aimed at determining the impact of SDF on fertility potentials in a rat model. Materials and Methods: Twenty adult male Sprague–Dawley (SD) rats were randomly divided into four groups of five rats each. Groups A1(distilled water) and B1 (2 g/kg of 30% v/v ethanol) lasted for 4 weeks while Groups A2(control; distilled water) and B2 (2 g/kg of 30% v/v ethanol) lasted for 8 weeks. At the end of each treatment, the animals were introduced to female SD rats on the proestrous day of their cycle. The testis was harvested and tested for oxidative stress while the cauda epididymis was harvested to test for epididymal sperm Parameters and SDF. Results: The sperm count, sperm motility, and the number of fetuses sired by the animals that received alcohol decreased significantly (P Conclusion: Alcohol Increased oxidative stress and SDF altering the ability of spermatozoa to fertilize oocytes.

Does sperm DNA fragmentation affect the developmental potential and the incidence of apoptosis following blastomere biopsy?

Common methods employed in assisted reproduction technology (ART) include intracytoplasmic sperm injection (ICSI) with an unspecified level of sperm DNA fragmentation (SDF) and preimplantation genetic diagnosis (PGD). The aim of this study was to investigate the impact of SDF on human preimplantation embryo development and the incidence of apoptosis following a single blastomere biopsy. Using sperm chromatin dispersion (SCD) to assess SDF, a total of 20 processed semen samples were categorized into two groups; group I: SDF ≤30% and group II: SDF >30%. After ICSI, fertilization, cleavage, and embryo quality score were assessed. A single blastomere was biopsied from day 3 embryos and development was monitored on day 4. The frequency of apoptosis in biopsied embryos was assayed by TUNEL and the level of BCL-2, BAX, hsa-mir-15a, and hsa-mir-16-1 were assessed by quantitative real-time polymerase chain reaction (qRT-PCR). SCD was found to be negatively correlated with sperm motility and normal form spermatozoa (p < 0.05). The rate of fertilization, cleavage, and embryo quality score were not significantly different between the two groups (all p > 0.05). SDF >30% had no negative effect on potential development and did not increase the proportion of apoptotic cells and the level of apoptosis-related genes and microRNAs (miRNAs) in group II vs. group I (p > 0.05). It appears that at the levels assessed paternal genome damage had little if any negative effect on preimplantaton embryo development and apoptosis following single blastomere biopsy. This may reflect the selection of morphologically normal sperm for ICSI and the repair capacity of the oocyte.