515 Sperm DNA integrity in patients with varicocele: relationships between DNA fragmentation and bulk semen parameters

SummarySperm DNA fragmentation is referred to as one of the main causes of male infertility. Failures in the protamination process, apoptosis and action of reactive oxygen species (ROS) are considered the most important causes of DNA fragmentation. Action of ROS or changes in sperm protamination would increase the susceptibility of sperm DNA to fragmentation. Routine semen analysis is unable to estimate sperm chromatin damage. Sperm DNA integrity influences sperm functional capability, therefore tests that measure sperm DNA fragmentation are important to assess fertility disorders. Actually, there is a considerable number of methods for assessing sperm DNA fragmentation and chromatin integrity, sperm chromatin stability assay (SCSA modified), sperm chromatin dispersion (SCD), comet assay, transferase dUTP nick end labelling (TUNEL); and protamine evaluation in sperm chromatin assay, such as toluidine blue, CMA3, protamine expression and evaluation of cysteine radicals. This review aims to describe the main causes of sperm DNA fragmentation and the tests commonly used to evaluate sperm DNA fragmentation.

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

Inflammatory Bowel Diseases ◽

10.1093/ibd/izab205 ◽

2021 ◽

Author(s):

Anne Grosen ◽

Emanuelle Bellaguarda ◽

Jacob Nersting ◽

Christian Lodberg Hvas ◽

Ingela Liljeqvist-Soltic ◽

...

Keyword(s):

Healthy Volunteers ◽

Dna Fragmentation ◽

Low Dose ◽

Inflammatory Diseases ◽

Methotrexate Therapy ◽

Semen Parameters ◽

Dna Integrity ◽

Sperm Dna Fragmentation ◽

Fragmentation Index ◽

Sperm Dna

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

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Acridine Orange and Flow Cytometry: Which Is Better to Measure the Effect of Varicocele on Sperm DNA Integrity?

Advances in Urology ◽

10.1155/2015/814150 ◽

2015 ◽

Vol 2015 ◽

pp. 1-6 ◽

Cited By ~ 9

Author(s):

Essam-Elden M. Mohammed ◽

Eman Mosad ◽

Asmaa M. Zahran ◽

Diaa A. Hameed ◽

Emad A. Taha ◽

...

Keyword(s):

Flow Cytometry ◽

Acridine Orange ◽

Pregnancy Outcome ◽

Dna Fragmentation ◽

Chromatin Condensation ◽

Semen Parameters ◽

Dna Integrity ◽

Sperm Dna Fragmentation ◽

Sperm Dna Damage ◽

Sperm Dna

We evaluated the effect of varicocelectomy on semen parameters and levels of sperm DNA damage in infertile men. A total of 75 infertile men with varicocele and 40 fertile men (controls) were included in this study. Semen analysis and sperm DNA damage expressed as the DNA fragmentation index using acridine orange staining and chromatin condensation test by flow cytometry were assessed before and 6 months after varicocelectomy. The patients were also followed up for 1 year for pregnancy outcome. Semen parameters were significantly lower in varicocele patients compared to controls (P<0.05). Mean percentages of sperm DNA fragmentation and sperm DNA chromatin condensation in patients were significantly higher than those in controls (P<0.05). After varicocelectomy, sperm DNA fragmentation improved significantly, whereas sperm chromatin condensation was not significantly changed. In 15 out of 75 varicocele patients, clinical pregnancy was diagnosed; those with positive pregnancy outcome had significant improvement in sperm count, progressive sperm motility, and sperm DNA fragmentation, but there was no significant difference in sperm DNA condensation compared to negative pregnancy outcome patients. We concluded from this study that acridine orange stain is more reliable method than flow cytometry in the evaluation of sperm DNA integrity after varicocelectomy.

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In Vitro Studying the Effect of Adding Autologous Platelet Rich Plasma (PRP) to the Human Semen on the Sperm DNA Integrity

Iraqi Journal of Embryos and Infertility Researches ◽

10.28969/ijeir.v10.i2.r7 ◽

2021 ◽

Vol 10 (2) ◽

pp. 90-100

Author(s):

Dhafer Hamdan ◽

Ali Rahim ◽

Ula Al-Kawaz

Keyword(s):

Dna Fragmentation ◽

Assisted Reproductive Technologies ◽

Platelet Rich Plasma ◽

Reproductive Technologies ◽

Human Reproduction ◽

Sperm Chromatin ◽

Dna Integrity ◽

Sperm Dna Integrity ◽

Sperm Dna ◽

Dispersion Test

For conception and the development of healthy embryos, sperm DNA integrity is crucial. According to a growing body of studies, there is a strong correlation between sperm DNA damage and male infertility. Among the new medicines being developed in the medical field, the application of Platelet Rich Plasma (PRP) in human reproduction has yet to be examined. A total of 100 semen samples were used in the current experimental investigation. From November 2020 to June 2021, the research was conducted at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies. Masturbation was used to get an ejaculated semen sample. After semen analysis, the samples were separated into two equal parts, one without autologous PRP and the other with 2% autologous PRP, with the DNA fragmentation assessed using the Sperm Chromatin Dispersion Test. There was highly significant reduction in DNA fragmentation index (p < 0.001). The mean sperm DNA integrity was reduced after adding PRP (33.85±16.73 vs 38.55±16.64), Mean (± SE). PRP has been shown to improve human sperm DNA integrity.

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1384: Evaluation of Sperm DNA Integrity, Standard Semen Parameters and Pregnancy Outcomes in Male Infertility

The Journal of Urology ◽

10.1016/s0022-5347(18)38609-9 ◽

2004 ◽

Vol 171 (4S) ◽

pp. 364-365

Author(s):

Yael Perez ◽

Jeanne H. O'Brien ◽

Sharon Sharir ◽

Keith A. Jarvi ◽

Armand Zini

Keyword(s):

Male Infertility ◽

Pregnancy Outcomes ◽

Semen Parameters ◽

Dna Integrity ◽

Sperm Dna Integrity ◽

Sperm Dna

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Are semen quality parameters sufficient for biomonitoring spermatozoa DNA integrity and oxidatively damaged DNA

Biomonitoring ◽

10.1515/bimo-2015-0004 ◽

2015 ◽

Vol 2 (1) ◽

Author(s):

Hueiwang Anna Jeng ◽

Ruei-Nian Li ◽

Wen-Yi Lin

Keyword(s):

Dna Damage ◽

Dna Fragmentation ◽

Semen Quality ◽

Quality Parameters ◽

Semen Parameters ◽

Sperm Chromatin ◽

Dna Integrity ◽

Phase System ◽

Sperm Dna Fragmentation ◽

Sperm Dna

Abstract:The present study aimed to investigate the relationship between semen quality parameters and DNA integrity, and determine whether semen quality parameters could serve as a reliable biomarker for monitoring sperm DNA damage. Conventional semen parameters from a total of 202 male human subjects were analyzed. DNA fragmentation and 8-oxo-7,8-dihydro-2′- deoxyguanosine (8-oxoGuo) were used to assess sperm DNA integrity. DNA fragmentation was analyzed by the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and sperm chromatin structure assay (SCSA), while 8-oxodGuo was quantified by the liquid chromatography/tandem mass spectrometry (LC-MS/MS) coupled with an on-line solid phase system. The levels of 8-oxodGuo levels in sperm were related to the percentages of DNA fragmentation measured by both the TUNEL and SCSA (r = 0.22, p = 0.048; r = 0.12, p = 0.039). Sperm vitality, motility and morphology from all of the participants exhibited a weak correlation with the levels of 8-oxodGuo and the percentages of DNA fragmentation. Semen quality parameters may be independent of the formation of DNA fragmentation and oxidative adducts in sperm. Semen quality parameters may be insufficient to monitor sperm DNA fragmentation and oxidative damage. DNA damage in sperm is recommended to be included in routine measurements.

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