Decreased Sperm DNA Fragmentation Research Articles

Second ejaculation produces good quality sperm and blastocyst and decreases the rate of unexpected ICSI cycle: a propensity score-matched analysis

BackgroundSecond ejaculation can influence sperm quality which may define the first-line treatment. The purpose of this study was to evaluate the effectiveness of a second ejaculation in decreasing the unexpected intracytoplasmic sperm injection (ICSI) rate by a propensity score-matched (PSM) analysis.MethodsPatients who were projected to undergo IVF were included between January 2016 and November 2021 in this monocentric, retrospective analysis. 2782 patients included in the study, 143 and 2639 patients were non-randomized in the unexpected ICSI and IVF groups, respectively. One hundred fourteen patients with unexpected ICSI produced two semen samples on the day of ovum pick-up. After 1:4 PSM, we matched 61 patients in the second ejaculation IVF group to 238 patients in the conventional IVF group. Outcomes of sperm quality, fertilization rate, embryo quality, and pregnancy were compared.ResultsSecond ejaculation significantly improved sperm concentration, progressive motility before and after sperm swim-up, total progressive motility sperm count after swim-up, and decreased sperm DNA fragmentation (SDF). Sixty-one of 114 (53.5%) unexpected ICSI couples had enough total progressive motility sperm for IVF with the second ejaculation. There were no differences in basic clinical characteristics between couples in second ejaculation IVF and matched-conventional IVF group. For the two groups, no differences were observed in IVF outcomes. However, a significant increase in good-quality blastocyst rate was observed for second-ejaculation IVF couples. Univariate and multivariate linear regression analysis also confirmed that the second ejaculation was an independent risk factor for the good quality blastocyst rate.ConclusionSecond ejaculation could be an economical and secure alternative to get good quality sperm, and blastocyst and decrease the rate of unexpected ICSI. Multicenter studies should be conducted to confirm the potential advantages of using second ejaculation IVF in effectively reducing the rate of ICSI.

Zishen Yutai Pill improves sperm quality and reduces testicular inflammation in experimental varicocele rats

ObjectiveZishen Yutai Pill (ZYP), containing 15 Chinese traditional medicine, is a safe and well quality-controlled TCM preparation with promising effects in many fields of reproduction. The current study was designed to investigate the therapeutic effects of ZYP on sperm quality and testis in varicocele (VC) rats. Materials and methodsMale Wistar rats were randomly divided into four groups (n = 6), i.e., a sham group, a VC group, and VC groups treated with different dose of ZYP (1575 and 3150 mg/kg/d, respectively). The experimental VC model was established by partial ligation of left renal vein. Six weeks after model establishment, ZYP was orally administered once a day for the next 6 weeks. Parameters relating to testis and sperm quality were assessed. Hematoxylin–eosin staining was used to showed testicular tissue damage in experimental VC rats. Expressions of proteins relating to NLRP3 inflammasome pathways were determined using Western blot (WB). The mRNA expressions of relating genes were determined using quantitative real-time PCR (qRT-PCR) analysis. ResultsZYP could significantly improve sperm motility and decrease sperm DNA fragmentation index in VC rats (P < 0.05). Hematoxylin-eosin (HE) staining showed that ZYP could alleviate testicular tissue damage caused by experimental varicocele in rats. Compared to the VC model, expressions of NLRP3, ASC, and caspase-1 in rats treated with ZYP were significantly down-regulated, as validated by both qRT-PCR and WB analysis (P < 0.05). ConclusionsIn brief, ZYP could improve sperm DNA integrity by inhibiting the NLRP3 inflammasome pathway and alleviating the chronic inflammation of testicular tissue induced by experimental varicocele in rats.

Efficacy of varicocelectomy for sperm DNA integrity improvement: A meta-analysis.

Varicocele is common among male adolescents and adults. Varicocelectomy is the major means of varicocele repair. There is evidence that varicocelectomy could decrease sperm DNA fragmentation. However, studies evaluating the efficacy of varicocelectomy for sperm DNA integrity usually have a small sample size, and there is no up-to-date meta-analysis in this area. The present meta-analysis is to evaluate the efficacy of varicocelectomy for sperm DNA integrity. A literature search was conducted to identify all relevant studies from Medline database (PubMed), Cochrane Library, and OVID Embase from the inception dates to 08 June 2020. A total of 11 prospective studies including 394 cases were included in this meta-analysis. All analysis was performed using Stata version 16.0. In the random-effect model for 11 studies, DNA fragmentation index (%) of clinical varicocele patients decreased by 5.79 (95% CI, -7.39 to -4.19) after varicocelectomy. While after excluding one study with high heterogeneity, from the results of fixed-effect model, DNA fragmentation index decreased by 6.14 (95% CI, -6.90 to -5.37) on average. Sperm DNA integrity of clinical varicocele patients was significantly improved following varicocelectomy. Therefore, it is necessary to include elevated sperm DNA fragmentation index as a molecular indicator for varicocelectomy among clinical varicocele cases.

Orlistat attenuates obesity‐induced decline in steroidogenesis and spermatogenesis by up‐regulating steroidogenic genes

Steroidogenesis decline is reported to be one of the mechanisms associated with obesity-induced male factor subfertility/infertility. We explored the possible preventive/therapeutic effects of orlistat (a medication prescribed for weight loss) on obesity-induced steroidogenesis and spermatogenesis decline. Twenty-four adult male Sprague Dawley rats weighing 250-300g were randomized into four groups (n=6/group), namely; normal control, high-fat diet, high-fat diet plus orlistat preventive group and high-fat diet plus orlistat treatment group. Orlistat (10mg/kg/b.w./d suspended in distilled water) was either concurrently administered with high-fat diet for 12weeks (high-fat diet plus orlistat preventive group) or administered from week 7-12 post- high-fat diet feeding (high-fat diet plus orlistat treatment group). Thereafter, serum, testes and epididymis were collected for analyses. Obesity increased serum leptin and decreased adiponectin levels, decreased serum and intra-testicular levels of follicle stimulating hormone, luteinising hormone and testosterone, sperm count, motility, viability, normal morphology and epididymal antioxidants, but increased epididymal malondialdehyde level and sperm nDNA fragmentation. Testicular mRNA transcript levels of androgen receptor, luteinizing hormone receptor, steroidogenic acute regulatory protein, cytochrome P450 enzyme (CYP11A1), 3β-hydroxysteroid dehydrogenase and 17β-hydroxysteroid dehydrogenase were significantly decreased in the testes of the high-fat diet group. Further, the levels of steroidogenic acute regulatory protein protein and enzymatic activities of CYP11A1, 3β-hydroxysteroid dehydrogenase and 17β-hydroxysteroid dehydrogenase were also significantly decreased in the testes of the high-fat diet group. Treatment with orlistat significantly decreased leptin and increased adiponectin levels, improved sperm parameters, decreased sperm DNA fragmentation, increased the levels of steroidogenic hormones, proteins and associated genes in high-fat diet-induced obese male rats, with the preventive group (high-fat diet plus orlistat preventive group) having better results relative to the treatment group (high-fat diet plus orlistat treatment group). Orlistat attenuated impaired spermatogenesis and steroidogenesis decline by up-regulating steroidogenic genes. This may not be unconnected to its significant effect in lowering serum leptin levels, since the hormone is known to dampen fertility potential. Therefore, orlistat may improve fertility potential in overweight/obese men.

Comparison between sperm parameters and chromatin in recurrent pregnancy loss couples after antioxidant therapy.

Background and Aims:Recurrent Pregnancy loss (RPL) is a heterogeneous disease. The role of maternal factor is clear but the relationship between the paternal factors remains uncertain. It has been shown that increase the level of Reactive Oxygen Species (ROS) and decrease the antioxidant levels in men can lead to RPL. New researches show treatment with antioxidant can improved sperm parameter. The aim of the present study was to assess the effect of vitamin E plus Zinc therapy on sperm parameters and chromatin quality in couples with RPL.Methods:In this clinical trial study, 60 RPL patients were selected from RPL clinic. Standard sperm parameters were analyzed and then male partners were intake vitamin E and Zinc in antioxidant therapy plan for 90 days. After that, sperm parameters were analyzed again. Sperm chromatin assay was reviewed before and after vitamin E and Zinc therapy by cytochemical assay including aniline blue (AB), chromomycin A3 (CMA3), toluidine blue (TB). To assess DNA fragmentation index, TUNEL test was used. Data were analyzed and compared before and after treatment.Results:data analysis showed all sperm parameters significantly improved after treatment (P < 0.001). The number of AB+ and TB+ sperms were decreased significantly after vitamin E and Zinc therapy (P = 0.0001). Decrease in DNA fragmentation in post treatment group in comparison to pre-treatment was statistically significant.Conclusion:Supplemental of vitamin E plus Zinc may improve sperm parameters chromatin quality and decrease sperm DNA fragmentation in RPL patients.

An update on clinical and surgical interventions to reduce sperm DNA fragmentation in infertile men

Sperm chromatin integrity is essential for normal embryo development and pregnancy outcome. Sperm DNA fragmentation (SDF) testing constitutes a diagnostic tool to measure the proportion of spermatozoa with damaged chromatin in the ejaculate. SDF is associated with potentially treatable conditions, including varicocele, male accessory gland infections, inadequate lifestyle, and gonadotoxin exposure, thus prompting their treatment as a means of improving sperm DNA quality and the reproductive outcomes. To provide an up-to-date review of the role of clinical and surgical interventions on SDF values in subfertile men. An extensive search of studies examining the relationship between male infertility conditions associated with SDF was performed using PubMed and MEDLINE, with a focus on interventional therapy. The start date for the search was not defined, whereas the end date was March 2019. Randomized and non-randomized controlled trials, observational studies, systematic and narrative reviews, and case series were evaluated. Treating the underlying male infertility factor seems a promising way to alleviate SDF and to increase the likelihood of achieving natural and assisted conception, but data remain limited. The best evidence relates to varicocele repair and hormonal therapy with the follicle-stimulating hormone. Antioxidant therapy and lifestyle changes might alleviate oxidative sperm markers and decrease SDF but their effects on pregnancy outcomes are still unclear. Among men with high SDF undergoing assisted reproductive technology, the use of testicular spermatozoa in preference over ejaculated spermatozoa for intracytoplasmic sperm injection (ICSI) has been shown to improve pregnancy rates possibly owing to the better sperm chromatin quality in testicular spermatozoa than in ejaculated spermatozoa. Current evidence supports interventional therapy as a means to alleviate sperm DNA damage. Identification of the conditions associated with SDF remains important to enable treatment to potentially improve pregnancy outcomes but given the limited data further research is needed to determine the exact role of specific interventional therapy for subfertile men with impaired sperm chromatin.

Measured with Sperm Chromatin Dispersion (SCD) Technique: Will DNA Integrity in Spermatozoa Increase After Micronutrient Supplementation? Comparative Controlled Study

Background: An elevated DNA Fragmentation Index (DFI) has been associated with male infertility; therefore measuring the sperm nuclear DNA integrity appears to be useful in predicting the ability of spermatozoa to fertilize oocytes.Aim: To evaluate the effect of micronutrient supplementation over DFI values in subfertile men.Methods: This was a retrospective/comparative study that included a total of 306 subfertile males consulting the clinic from March 2011 to November 2017, who had performed the Sperm Chromatin Dispersion test (SCD) — a method used to detect DNA fragmentation — along with the initial semen analysis. Of the included population, [Formula: see text] 146 had received two daily oral capsules of a standardized combined micronutrient compound ([Formula: see text], Lenus Pharma, Vienna, Austria) for three months plus lifestyle change (study group). Each capsule contains L-carnitine, L-arginine, coenzyme Q10, zinc, vitamin E, folic acid, glutathione and selenium. Those who did not receive the active treatment ([Formula: see text] 160) but only engaged with lifestyle changes were considered controls. The SCD test was performed in both groups at baseline and after 3 months.Results: For the first statistical analysis, patients with an initial mean DFI of &gt;15% were considered ([Formula: see text] 66 [37 were study and 29 were controls]). After 3 months, both groups displayed a significant decrease of mean DFI values. However, this decrease was more evident in the study group as compared to controls (10.54% vs. 14.48%, [Formula: see text] 0.05; [Formula: see text] 0.013). For the second statistical approach the entire population was considered ([Formula: see text] 306). After 3 months, only the study group displayed a significant decrease of the mean initial DFI value (10.16% to 6.49%, [Formula: see text] &lt; 0.0001); decrease that was more evident as compared to controls (6.49% vs. 8.82%, [Formula: see text] 0.05; [Formula: see text] 0.000020).Conclusions: Among subfertile men with a DFI &gt;15%, both regimes significantly decreased sperm DNA fragmentation; however, when any baseline DFI value was considered, only treatment with the active standardized micronutrient compound achieved a significant better result.

Use of microfluidic sperm extraction chips as an alternative method in patients with recurrent in vitro fertilisation failure.

It is known that sperm preparation techniques in in vitro fertilisation (IVF) are intended to select the best-quality sperm. The aim of this study is to compare sperm the density gradient method and microfluidic chip (Fertile Plus) method in infertile patients by analysing fertilisation rates, pregnancy rates, and sperm morphology and DNA fragmentation rates posed by these two methods. Using semen samples obtained from the patients, sperms were prepared with gradient (n = 312) and microfluidic chip methods (n = 116). Fertilisation and pregnancy rates were compared in the first time and in the recurrent IVF trial patients. In addition, the morphology and DNA fragmentation comparison of sperm samples were evaluated by Toluidine blue in situ chemical staining method. There was no statistically significant difference between fertilisation and pregnancy rates when compared with study groups in first-time IVF treatment patients. However, in recurrent IVF failure patients, there was a significant difference in fertilisation rates but no statistically significant difference was found in pregnancy rates. The microfluidic chip method significantly decreased sperm DNA fragmentation index according to density gradient method. Microfluidic chip method may be recommended in patients with recurrent unsuccessful in vitro trials. The sperm DNA fragmentation test prior to the treatment will be helpful in selecting the appropriate sperm-washing method.

Effect of Lepidium meyenii (maca) on testicular function of mice with chemically and physically induced subfertility.

The aim of this study was to evaluate the effect of Lepidium meyenii (maca) in chemically and physically subfertile mice. After 35days, the following groups of mice were evaluated: control, sham, chemical subfertility, chemical subfertility-maca-supplemented, physical subfertility, physical subfertility-maca-supplemented and maca-supplemented only. Motility (32.36%±5.34%) and sperm count (44.4±5.37×10(6) /ml) in the chemically and physically subfertile mice (11.81%±4.06%, 17.34±13.07×10(6) /ml) decreased compared to the control (75.53%±2.97% and 57.4±19.6 10(6) /ml) and sham (53.5%±7.86% and 58.4±14.10 10(6) /ml). Maca was able to reverse the deleterious effect of motility (76.36±1.97) as well as sperm count (53.5±9.18×10(6) /ml) on chemical subfertility. In contrast, maca did not reverse the effects of induced physical subfertility nor motility (18.78%±14.41%) or sperm count (20.17±11.20×10(6) /ml). The percentage of sperm DNA fragmentation in the physically subfertile mice increased (11.1%±19.29%) compared to the control (0.84%±0.85%). However, in the physically subfertile group, maca decreased sperm DNA fragmentation (2.29%±2.30%) closer to the sham (1.04%±0.62%) and the control (0.84%±0.85%). The group supplemented only with maca showed 0.54%±0.50% of spermatozoa with DNA fragmentation. Yet, the differences observed were statistically not significant. In conclusion, it appears that maca activates the cytochrome P450 system after chemically induced subfertility. However, it does not reverse the low mitochondrial membrane potential in spermatozoa compromised in the physical subfertility group.

Effect of genistein added to bull semen after thawing on pronuclear and sperm quality

The aim of this research was to study the effect of different genistein treatments on bull sperm after thawing on pronuclear formation after in vitro fertilization (IVF) and on different sperm quality variables. Three experiments were performed. In Experiment 1, three treatments (Control, sperm incubation for 1h at 37°C with or without genistein) and two sperm concentrations during IVF (1 or 3×106sperm/mL) were evaluated to study the influence of genistein on pronuclear formation (PNF). Sperm incubation for 1h before IVF reduced PNF regardless of sperm concentration. However, after sperm incubation and with 3×106sperm/mL in IVF, the genistein treatment group had greater fertilization rates than the untreated group. In Experiment 2, six treatments plus the control group were performed to study the effect of genistein (presence or not) and incubation conditions (30min at 37°C, 1h at 27°C or at 37°C) on PNF using 3×106sperm/mL for IVF. When incubation time was reduced to 30min, PNF rate from the genistein treatment group was no different from either the control group or in the group in which incubation occurred for 1h at 27°C. In Experiment 3, the effect of several genistein treatments (control; genistein treatment for 30min of incubation at 37°C; genistein treatment for 1h of incubation at 27°C) on sperm motility, viability and DNA fragmentation were evaluated. Genistein did not improve sperm motility and, depending on the experimental group or time, it either reduced or had no effect on sperm motility. Genistein treatment did not improve sperm viability after 5h of incubation. However, genistein treatment for 1h at 27°C decreased sperm DNA fragmentation compared with the control group after 5h of sperm incubation. In conclusion, the treatment of bull sperm with genistein for 1h at 27°C could decrease sperm DNA fragmentation, although PNF rate after IVF and sperm motility were reduced.

Decreased Sperm DNA Fragmentation After Surgical Varicocelectomy is Associated With Increased Pregnancy Rate

We prospectively evaluated changes in sperm chromatin structure in infertile patients before and after surgical repair of varicocele, and the impact on the pregnancy rate. Included in the study were 49 men with at least a 1-year history of infertility, a palpable varicocele and oligospermia. World Health Organization semen analysis and sperm DNA damage expressed as the DNA fragmentation index using the sperm chromatin structure assay were assessed preoperatively and postoperatively. Pregnancy (spontaneous and after assisted reproductive technique) was recorded 2 years after surgery. Mean sperm count, sperm concentration and sperm progressive motility improved significantly after varicocelectomy from 18.3 × 10(6) to 44.4 × 10(6), 4.8 × 10(6)/ml to 14.3 × 10(6)/ml and 16.7% to 26.6%, respectively (p <0.001). The DNA fragmentation index decreased significantly after surgery from 35.2% to 30.2% (p = 0.019). When the definition of greater than 50% improvement in sperm concentration after varicocelectomy was applied, 31 of 49 patients (63%) responded to varicocelectomy. After varicocelectomy 37% of the couples conceived spontaneously and 24% achieved pregnancy with assisted reproductive technique. The mean postoperative DNA fragmentation index was significantly higher in couples who did not conceive spontaneously or with assisted reproductive technique (p = 0.033). After varicocelectomy sperm parameters significantly improved and sperm DNA fragmentation was significantly decreased. Low DNA fragmentation index values are associated with a higher pregnancy rate (spontaneous and with assisted reproductive technique). We suggest that varicocelectomy should be considered in infertile men with palpable varicocele, abnormal semen analysis and no major female factors.

Reduction of increased sperm DNA fragmentation index using low-dose clomiphene citrate treatment among infertile males

Increased sperm DNA fragmentation is associated with reduced reproductive outcome. Varicocelectomy and anti-oxidant treatments were reported to decrease sperm DNA fragmentation index (DFI). Clomiphene citrate (CC) has been used to improved sperm quality, but its effect on sperm DFI is unknown. Effect of CC on sperm DFI was longitudinally evaluated.

Decreased Sperm DNA Fragmentation After Surgical Varicocelectomy is Associated With Increased Pregnancy Rate

We prospectively evaluated changes in sperm chromatin structure in infertile patients before and after surgical repair of varicocele, and the impact on the pregnancy rate. Included in the study were 49 men with at least a 1-year history of infertility, a palpable varicocele and oligospermia. World Health Organization semen analysis and sperm DNA damage expressed as the DNA fragmentation index using the sperm chromatin structure assay were assessed preoperatively and postoperatively. Pregnancy (spontaneous and after assisted reproductive technique) was recorded 2 years after surgery. Mean sperm count, sperm concentration and sperm progressive motility improved significantly after varicocelectomy from 18.3 x 10(6) to 44.4 x 10(6), 4.8 x 10(6)/ml to 14.3 x 10(6)/ml and 16.7% to 26.6%, respectively (p <0.001). The DNA fragmentation index decreased significantly after surgery from 35.2% to 30.2% (p = 0.019). When the definition of greater than 50% improvement in sperm concentration after varicocelectomy was applied, 31 of 49 patients (63%) responded to varicocelectomy. After varicocelectomy 37% of the couples conceived spontaneously and 24% achieved pregnancy with assisted reproductive technique. The mean postoperative DNA fragmentation index was significantly higher in couples who did not conceive spontaneously or with assisted reproductive technique (p = 0.033). After varicocelectomy sperm parameters significantly improved and sperm DNA fragmentation was significantly decreased. Low DNA fragmentation index values are associated with a higher pregnancy rate (spontaneous and with assisted reproductive technique). We suggest that varicocelectomy should be considered in infertile men with palpable varicocele, abnormal semen analysis and no major female factors.

Development and performances of plastic microfluidic sperm sorter

OBJECTIVE: Microfluidic sperm sorter (MFSS) is a simple chip device to select motile sperm for ART, in which sperms are selected by two gravity-driven laminar flows within the microfluidic channel. Conventional semen processing techniques such as density gradient and serial centrifugation can have potential adverse effects on sperm DNA integrity. A present report suggests that MFSS decrease sperm DNA fragmentation while significantly increased mean sperm motility compared to the unprocessed sample.

Attenuation of Oxidative Stress After Varicocelectomy in Subfertile Patients With Varicocele

We examined changes due to oxidative damage to spermatozoa and alterations in antioxidant capacity in subfertile patients with varicocele before and after varicocelectomy in a prospective study. A total of 30 young subfertile male patients with varicocele were recruited in this study. Varicocele was diagnosed by physical examination and Doppler ultrasound. Semen analysis was performed in the 30 patients before and 6 months after varicocelectomy using a computer assisted semen analyzer. The parameters for evaluating oxidative stress changes were 4977 bp deletion of mitochondrial DNA in sperm, as detected by polymerase chain reaction, the 8-OHdG (8-hydroxy-2'-deoxyguanosine) content in spermatozoa DNA, as measured by a high performance liquid chromatography electrochemical method, and seminal plasma protein thiols and ascorbic acid, as measured by spectrophotometric methods. Semen quality, including motility, morphology and sperm density, was improved in 22 patients (73.3%) after varicocelectomy. The incidence of 4977 bp deletion of mitochondrial DNA in sperm was 40% (12 of 30 patients) and 13.3% (4 of 30) before and after surgery, respectively. Mean +/- SD 8-OHdG content in sperm DNA, and seminal plasma protein thiols and ascorbic acid were 10.27 +/- 2.24/10(5) 2'-deoxyguanosine, 0.77 +/- 0.75 nmole/ml and 1.87 +/- 0.40 mg/dl before operation, and 5.95 +/- 1.46/10(5) 2'-deoxyguanosine, 3.00 +/- 1.17 nmole/ml and 3.12 +/- 0.94 mg/dl after surgery, respectively. The incidence of 4977 bp deletion of mitochondrial DNA in sperm and the level of 8-OHdG in sperm DNA were decreased, and seminal plasma protein thiols and ascorbic acid were increased significantly in all 30 patients after varicocelectomy. Also, in the 8 patients in whom semen quality did not improve after surgery a significant decrease in 8-OHdG in sperm DNA, and a significant increase in seminal plasma protein thiols and ascorbic acid were observed. Subfertile patients with varicocele had a significant decrease in oxidative damage in sperm DNA and an increase in antioxidant capacity in seminal plasma after varicocelectomy, indicating that surgery is effective treatment in such patients.