Background:A high level of sperm chromatin decondensation results in loss of DNA quality and it is associated with poor embryonic prognosis in Assisted Reproductive Technology (ART). This had lead to introduction of sperm DNA compaction testing in assessment of clinical male infertility. Toluidine blue assay is one of most commonly used tests to measure chromatin decondensation in research studies and also in clinical routine use. So the objective of our study was to assess the level of sperm chromatin decondensation by toluidine blue assay in men dealing with couple infertility. Methods: The sperm samples of 96 patients referred for infertility, over a 4-month period from September 2020 to January 2021, were stained with toluidine blue and observed under an optical microscope. These were evaluated according to the World Health Organization. Results: The 25-34 age group was the most represented among infertile men. There was a non-significant correlation between age and the level of spermatic chromatin condensation (r = 0.0348). Spermatic parameters had averages above standards norms, apart from that of low vitality. Spermatic chromatin decondenization affected 96% of infertile patients. Asthenozoospermia was the most common abnormality. Among standard parameters of spermogram, only the concentration of spermatozoa in the semen was weakly correlated (r = 0.18) with sperm chromatin decondensation. Conclusion: The level of sperm chromatin condensation is, on the one hand, a parameter independent of the age of the subject;？on the other hand, allows to evaluate male infertility, like the classic parameters of the spermogram and spermocytogram. However, in view of the high probability of changes in the nuclear quality of sperm intrinsic to the
References
[1]
Zegers-Hochschild, F., Adamson, G.D., De Mouzon, J., Ishihara, O., Mansour, R., Nygren, K., et al. (2009) The International Committee for Monitoring Assisted Reproductive Technology (ICMART) and the World Health Organization (WHO) Revised Glossary on ART Terminology, 2009. Human Reproduction, 24, 2683-2687.
https://doi.org/10.1093/humrep/dep343
[2]
Kliesch, S. (2014) Diagnosis of Male Infertility: Diagnostic Work-Up of the Infertile Man. European Urology Supplements, 13, 73-82.
https://doi.org/10.1016/j.eursup.2014.08.002
[3]
Meacham, R.B., Joyce, G.F., Wise, M., Kparker, A. and Niederberger, C. (2007) Male Infertility. Journal of Urology 177, 2058-2066.
https://doi.org/10.1016/j.juro.2007.01.131
[4]
Skakkebaek, N.E., Giwercman, A. and de Kretser, D. (1994) Pathogenesis and Management of Male Infertility. Lancet, 343, 1473-1479.
https://doi.org/10.1016/S0140-6736(94)92586-0
[5]
Agarwal, A., Mulgund, A., Hamada, A. and Chyatte, M.R. (2015) A Unique View on Male Infertility around the Globe. Reproductive Biology and Endocrinology, 13, Article No. 37. https://doi.org/10.1186/s12958-015-0032-1
[6]
Owolabi, A.T., Fasubaa, O.B. and Ogunniyi, S.O. (2013) Semen Quality of Male Partners of Infertile Couples in Ile-Ife, Nigeria. Nigerian Journal of Clinical Practice, 16, 37-40. https://doi.org/10.4103/1119-3077.106729
[7]
De la Rochebrochard, E. and Thonneau, P. (2002) Paternal Age and Maternal Age Are Risk Factors for Miscarriage; Results of a Multicentre European Study. Human Reproduction, 17, 1649-1656. https://doi.org/10.1093/humrep/17.6.1649
[8]
Kim, H.S., Kang, M.J., Kim, S.A., Oh, S.K., Kim, H., Ku, S.Y., et al. (2013) The Utility of Sperm DNA Damage Assay Using toluIdine Blue and Aniline Blue Staining in Routine Semen Analysis. Clinical and Experimental Reproductive Medicine, 40, 23-28. https://doi.org/10.5653/cerm.2013.40.1.23
[9]
British Andrology Society (BAS) (1999) British Andrology Society Guidelines for the Screening of Semen Donors for donor Insemination (1999). Human Reproduction, 14, 1823-1826. https://doi.org/10.1093/humrep/14.7.1823
[10]
Aoki, V.W., Emery, B.R., Liu, L. and Carrell, D.T. (2006) Protamine Levels Vary between Individual Sperm Cells of Infertile Human Males and Correlate with Viability and DNA Integrity. Journal of Andrology, 27, 890-898.
https://doi.org/10.2164/jandrol.106.000703
[11]
Sharma, R.K., Sabanegh, E., Mahfouz, R., Gupta, S., Thiyagarajan, A. and Agarwal, A. (2010) TUNEL as a Test for Sperm DNA Damage in the Evaluation of Male Infertility. Urology, 76, 1380-1386. https://doi.org/10.1016/j.urology.2010.04.036
[12]
Simon, L. and Lewis, L.E. (2011) Sperm DNA Damage or Progressive Motility: Which on Is Better Predictor of Fertilization in Vitro? Systems Biology in Reproductive Medicine, 57, 133-138. https://doi.org/10.3109/19396368.2011.553984
[13]
American Society for Reproductive Medicine (ASRM) (1998) Guidelines for Gamete and Embryo Donation. Fertility and Sterility, 70, 1S-13S.
[14]
De la Rochebrochard, E. and Thonneau, P. (2003) Paternal Age over 40 Years: An Important Risk Factor for Infertility. American Journal of Obstetrics & Gynecology, 189, 901-905. https://doi.org/10.1067/S0002-9378(03)00753-1
[15]
Dunson, D.B., Colombo, B. and Baird, D.D. (2002) Changes with Age in the Level and Duration of Fertility in the Menstrual Cycle. Human Reproduction, 17, 1399- 1403. https://doi.org/10.1093/humrep/17.5.1399
[16]
Watanabe, Y., Cornet, D., Merviel, P., Mandelbaum, J., Antoine, J.M. and Uzan, S. (2000) Influence of Husband’s Age on Outcome of a Shared Oocyte Donation Program. Fertility and Sterility, 74, S78-S79.
https://doi.org/10.1016/S0015-0282(00)00933-X
[17]
Astolfi, P., De Pasquale, A. and Zonta, L.A. (2004) Late Paternity and Stillbirth Risk. Human Reproduction, 19, 2497-2501. https://doi.org/10.1093/humrep/deh449
[18]
Gandini, L., Lombardo, F., Paoli, D., Caponecchia, L., Familiari, G., Verlengia, C., et al. (2000) Study of Apoptotic DNA Fragmentation in Human Spermatozoa. Human Reproduction, 15, 830-839. https://doi.org/10.1093/humrep/15.4.830
[19]
Zini, A., Bielecki, R., Phang, D. and Zenzes, M.T. (2001) Correlations between two Markers of Sperm DNA Integrity, DNA Denaturation and DNA Fragmentation, in Fertile and Infertile Men. Fertility and Sterility, 75, 674-677.
https://doi.org/10.1016/S0015-0282(00)01796-9
[20]
Marchetti, C., Obert, G., Defosez, A., Formstecher, P. and Marchetti, P. (2002) Study of Mitochondrial Membrane Potential, Reactive Oxygen Species, DNA Fragmentation and Cell Viability by Flow Cytometry in Human Sperm. Human Reproduction, 17, 1257-1265. https://doi.org/10.1093/humrep/17.5.1257
[21]
Benchaib, M., Braun, V., Lornage, J., Hadj, S., Salle, B., Lejeune, H., et al. (2003) Sperm DNA Fragmentation Decreases the Pregnancy Rate in an Assisted Reproductive Technique. Human Reproduction, 18, 1023-1028.
https://doi.org/10.1093/humrep/deg228
[22]
Liu, C.H., Tsao, H.M., Cheng, T.C., Wu, H.M., Huang, C.C., Chen, C.I., et al. (2004) DNA Fragmentation, Mitochondrial Dysfunction and Chromosomal Aneuploidy in the Spermatozoa of Oligoasthenoteratozoospermic Males. Journal of Assisted Reproduction and Genetics, 21, 119-126.
https://doi.org/10.1023/B:JARG.0000029495.22787.83
[23]
Huang, C.C., Lin, D.P., Tsao, H.M., Cheng, T.C., Liu, C.H. and Lee, M.S. (2005) Sperm DNA Fragmentation Negatively Correlates with Velocity and Fertilization Rates But Might Not Affect Pregnancy Rates. Fertility and Sterility, 84, 130-140.
https://doi.org/10.1016/j.fertnstert.2004.08.042
[24]
Piasecka, M., Gaczarzewicz, D., Laszczynska, M., Starczewski, A. and Brodowska, A. (2007) Flow Cytometry Application in the Assessment of Sperm DNA Integrity of Men with Asthenozoospermia. Folia Histochemica et Cytobiologica, 45, S127-S136.
[25]
Varum, S., Bento, C., Sousa, A.P., Gomes-Santos, C.S., Henriques, P., Almeida- Santos, T., et al. (2007) Characterization of Human Sperm Populations Using Conventional Parameters, Surface Ubiquitination, and Apoptotic Markers. Fertility and Sterility, 87, 572-583. https://doi.org/10.1016/j.fertnstert.2006.07.1528
[26]
Tognifode, M.V., Lokossou, S., Aboubakar, M., Ogoudjobi, M., Hounkpatin, B., Tonato-Bagnan, A., et al. (2018) Paramètres du spermogramme dans l’infertilité du couple: Cas d’une clinique privée du Bénin a propos de 210 cas. Journal de la Société de Biologie Clinique du Bénin, No. 28, 41-45.
[27]
Ajina, T., Ammar, O., Haouas, Z., Sallem, A., Ezzi, L., Grissa, I., et al. (2017) Assessment of Human Sperm DNA Integrity Using Two Cytochemical Tests: Acridine Orange Test and Toluidine Blue Assay. Andrologia, 49, Article ID: e12765.
https://doi.org/10.1111/and.12765
[28]
Pourmasumi, S., Khoradmehr, A., Rahiminia, T., Sabeti, P., Talebi, A.R., Ghasemzadeh, J. (2019) Evaluation of Sperm Chromatin Integrity Using Aniline Blue and Toluidine Blue Staining in Infertile and Normozoospermic Men. Journal of Reproduction & Infertility, 20, 95-101
[29]
Hammadeh, M.E., Zeginiadov, T., Rosenbaum, P., Georg, T., Schmidt, W. and Strehler, E. (2001) Predictive Value of Sperm Chromatin Condensation (Aniline Blue Staining) in the Assessment of Male Fertility. Archives of Andrology, 46, 99-104.
https://doi.org/10.1080/01485010117363
[30]
Henkel, R., Bastiaan, H.S., Schüller, S., Hoppe, I., Starker, W. and Menkveld, R. (2010) Leucocytes and Intrinsic ROS Production May Be Factors Compromising Sperm Chromatin Condensation Status. Andrologia, 42, 69-75.
https://doi.org/10.1111/j.1439-0272.2009.00967.x
[31]
Balhorn, R. (1982) A Model for the Structure of Chromatin in Mammalian Sperm. Journal of Cell Biology, 93, 298-305. https://doi.org/10.1083/jcb.93.2.298
[32]
Agarwal, A. and Said, T.M. (2018) Sperm Chromatin Assessment. In: Textbook of Assisted Reproductive Technique, 2nd Edition, 93-106.
[33]
Erenpreisa, J., Erenpreiss, J., Freivalds, T., Slaidina, M., Krampe, R., Butikova, J., et al. (2003) Toluidine Blue Test for Sperm DNA Integrity and Elaboration of Image Cytometry Algorithm. Cytometry Part A, 52A, 19-27.
https://doi.org/10.1002/cyto.a.10015
