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

Author(s):  
Anne Grosen ◽  
Emanuelle Bellaguarda ◽  
Jacob Nersting ◽  
Christian Lodberg Hvas ◽  
Ingela Liljeqvist-Soltic ◽  
...  

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.

2010 ◽  
Vol 22 (1) ◽  
pp. 312 ◽  
Author(s):  
M. Hidalgo ◽  
M. R. Murabito ◽  
M. J. Gálvez ◽  
S. Demyda ◽  
L. J. De Luca ◽  
...  

Recently, a new procedure for the analysis of sperm DNA fragmentation has been developed for humans and different mammalian species, using a commercial kit based on the sperm chromatin dispersion (SCD) test; however, a descriptive study in canine semen has not been performed. The aim of this work was to assess the sperm DNA fragmentation in canine ejaculates using the SCD test and 2 different staining techniques. For this purpose, ejaculates were collectedby digital manipulation from4 healthy dogs of different breeds (1 German Pointer, 2 Spanish Greyhounds, and 1 Crossbreed). After collection, the sperm-rich fraction of the ejaculates from 3 dogs were pooled each time (n = 4) and then extended in Dulbecco’s phosphate buffered saline. All the pooled semen samples presented physiological values concerning routine semen parameters (motility, morphology, and sperm concentration). The sperm DNA fragmentation was assessed using the Sperm-Halomax® commercial kit specifically developed for canine semen (Halotech DNA SL, Madrid, Spain). Two semen aliquots of the diluted pooled semen samples were processed on each pre-treated slide provided in the kit following the manufacturer’s instructions. The last step was the staining technique. We stained each slide with 2 different staining procedures. The first half of the slide was stained with propidium iodide (Sigma-Aldrich, St. Louis, MO, USA) mixed in a proportion 1 : 1 with an antifading solution. The second half of the slide was stained for 15 min in Wright solution (1.01383.0500, Merck, Whitehouse Station, NJ, USA) 1 :1 in Phosphate Buffer pH 6.88 (1.07294.1000, Merck). The stained slides were observed using fluorescence and light microscopy, respectively. Five hundred sperm per slide were counted. Spermatozoa with fragmented DNA showed a large and spotty halo of chromatin dispersion. Unfragmented sperm only showed a small and compact halo. Statistical analyses were performed using the Statistical Package for Social Science version 12.0 (SPSS Inc., Chicago, IL, USA). The sperm DNA fragmentation index was compared between Wright and fluorescence staining methods by ANOVA. Results were expressed as mean ± standard error of the mean. The first report of the sperm DNA fragmentation index in canine ejaculates was 2.26 ± 0.53% for Wright staining and 1.99 ± 0.10% for fluorescence technique. No differences were found between staining procedures. In conclusion, it was possible to assess the sperm DNA fragmentation of canine ejaculates using 2 different staining procedures, expecting that continuous research could be useful in defining the role of DNA fragmentation SCD test in canine semen evaluation and cryopreservation.


2015 ◽  
Vol 61 (5) ◽  
pp. 48-55 ◽  
Author(s):  
Irina Ivanovna Vitiazeva ◽  
Marina Victorovna Altashina ◽  
Tatiana Vladimirovna Mun ◽  
Ekaterina Anatolievna Troshina

The reduction of the birth rates in the developed countries and increase in the frequency of male infertility stimulate the extensive investigations for the factors that negatively affect the reproductive system of the men and causing their infertility. The excessive body weight and obesity in the men of the reproductive age can promote the development of infertility. One of the mechanisms by which excess fat tissue has a negative impact on male fertility is disturbance of spermatogenesis. The authors aggregate scientific publications concerning the macroscopic and ultrastructural disturbances of spermatogenesis in men with obesity. We present the results of the study conducted at the Department of ART Endocrinology Research Center, targeted at the revelation of the relationship of body mass index of men of reproductive age, semen parameters, sperm DNA fragmentation index, as well as the influence of body mass index on outcomes of in vitro fertilization programs.


2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Essam-Elden M. Mohammed ◽  
Eman Mosad ◽  
Asmaa M. Zahran ◽  
Diaa A. Hameed ◽  
Emad A. Taha ◽  
...  

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.


Biomonitoring ◽  
2015 ◽  
Vol 2 (1) ◽  
Author(s):  
Hueiwang Anna Jeng ◽  
Ruei-Nian Li ◽  
Wen-Yi Lin

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.


2019 ◽  
Vol 91 (2) ◽  
Author(s):  
Viktor Alargkof ◽  
Larissa Kersten ◽  
Romil Stanislavov ◽  
Zdravko Kamenov ◽  
Panagiotis Nikolinakos

Objective: This exploratory retrospective study aimed to compare the level of Sperm DNA Fragmentation (SDF) and investigate its association with bulk semen parameters, for the first time in Bulgarian patients with varicocele, using a distinct methodology. Material and methods: Standard semen analysis was performed according to the 2010 criteria of the European Society of Human Reproduction and Embryology - Nordic Association for Andrology (ESHRE-NAFA-2010) and DNA fragmentation was assessed using the Halosperm® kit. The total sample included 28 males: the control group consisted of men with normal genital examination and unknown fertility (n = 10), group one consisted of men with varicocele, normozoospermia and DNA fragmentation > 15% (n = 9) and group two consisted of men with varicocele, abnormal sperm parameters and DNA fragmentation > 15% (n = 9). Results: DNA fragmentation was found to be higher in patients with abnormal sperm parameters (43.78 ± 30.78) compared to the normozoospermic group (21.22 ± 3.93) (p = 0.008). In normozoospermic patients, no statistically significant correlations were observed between SDF and bulk semen parameters. In patients with abnormal sperm parameters, DNA fragmentation exhibited significant very strong negative association with motility (a+b), vitality and typical morphology (p < 0.001). Conclusions: DNA integrity assays could be used for a better evaluation and management of male infertility, particularly in normozoospermic varicocele patients.


Zygote ◽  
2021 ◽  
pp. 1-4
Author(s):  
Caixia Yuan ◽  
Haixia Song ◽  
Zhulin Wang ◽  
Huaixiu Wang

Summary Intact human sperm DNA is an essential prerequisite for successful fertilization and embryo development. Abnormal sperm DNA fragmentation is a independent factor for male infertility. The objective of this study was to investigate the effects of Peijingsu, a health product, on the DNA integrity of human sperm. Peijingsu was administered for 15 days to 22 patients who had an abnormal sperm DNA fragmentation index (DFI). The DFIs before and after treatment were compared and analyzed using paired t-test. DFIs decreased significantly (P = 0.0008) after treatment, therefore it was concluded that Peijingsu effectively improved sperm DNA integrity in infertile patients who had an abnormal sperm DFI.


Author(s):  
Láyonal Germán Acosta Campos ◽  
Lissett Chiscul Requejo ◽  
Carlos Antonio Rivas Miñano ◽  
Jheny Díaz Orrego ◽  
Elmer Chávez Loyaga ◽  
...  

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