The relationship between DNA fragmentation and the intensity of morphologically abnormal human spermatozoa

Abstract

Objective: To determine the relationship between teratozoospermia and sperm DNA fragmentation (SDF) in the human ejaculate.

Methods: This retrospective study included 100 normozoospermic men as a control cohort (abnormal forms >14%), 210 patients with a high level of abnormal forms (≤4%) and 65 patients presenting with a moderate level of abnormal forms (>4% to ≤14%) based on the World Health Organization definitions. Sperm morphology was assessed using bright field microscopy. Sperm DNA fragmentation was assessed using the sperm chromatin dispersion assay. Non-parametric analyses were conducted to examine the relationship between abnormal sperm morphology and sperm DNA fragmentation; receiver operating characteristic (ROC) analyses were conducted to assess sensitivity and specificity of this relationship.

Results: A correlation analysis revealed that the higher the proportion of abnormal spermatozoa in the ejaculate, the higher the level of SDF (Spearman's Rho = -0.230; P<0.001). Significant differences in the proportion of SDF were found when all cohorts were compared (P<0.001); these significant differences were also retained when the different cohorts were compared pairwise. ROC analysis showed a moderate but significant predictive value for SDF to differentiate patients with different levels of teratozoospemia.

Conclusions: Although analysis of a more continuous range of values for teratozoospermia would help further clarify any causal relationship with SDF, there is clearly a synergistic or coincident affiliation between these variables that needs to be acknowledged by the clinician when interpreting the spermiogram.

Keywords

Teratozoospermia / Sperm morphology / Sperm DNA fragmentation / Male factor / Human reproduction / Human fertility

Conflict of interest statement

The authors declare no conflicts of interest.

Funding

This study received no extramural funding.

Authors' contributions

Mercedes González-Martínez performed sperm DNA fragmentation analysis and data searching. Pascual Sánchez-Martín carried out patient recruitment and study design. Carmen López-Fernández performed sperm DNA fragmentation analysis. Stephen D. Johnston was responsible for data interpretation, draft and final manuscript writing. Jaime Gosálvez performed study design, data analysis and first draft production.

Publisher's Note

The Publisher of the Journal remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

References

[1] De Braekeleer M, Nguyen MH, Morel F, Perrin A. Genetic aspects of monomorphic teratozoospermia: A review. J Assist Reprod Genet 2015; 32:615-623. doi: 10.1007/s10815-015-0433-2.

[2] Danis RB, Samplaski MK. Sperm morphology: History, challenges, and impact on natural and assisted fertility. Curr Urol Rep 2019; 20:43. doi: 10.1007/s11934-019-0911-7.

[3] Franken DR. How accurate is sperm morphology as an indicator of sperm function? Andrologia 2015; 47:720-723. doi: 10.1111/and.12324.

[4] Linn E, Ghanem L, Bhakta H, Greer C, Avella M. Genes regulating spermatogenesis and sperm function associated with rare disorders. Front Cell Dev Biol 2021; 9: 634536. doi: 10.3389/fcell.2021.634536.

[5] Sivanarayana T, Krishna ChR, Prakash GJ, Krishna KM, Madan K, Rani BS, et al. CASA derived human sperm abnormalities: Correlation with chromatin packing and DNA fragmentation. J Assist Reprod Genet 2012; 29:1327-1334. doi: 10.1007/s10815-012-9885-9.

[6] Daris B, Goropevsek A, Hojnik N, Vlaisavljevi V. Sperm morphological abnormalities as indicators of DNA fragmentation and fertilization in ICSI. Arch Gynecol Obstet 2010; 281:363-367. doi: 10.1007/s00404-009-1140-y.

[7] Mangiarini A, Paffoni A, Restelli L, Ferrari S, Guameri C, Ragni G, et al. Specific sperm defects are differentially correlated with DNA fragmentation in both normozoospermic and teratozoospermic subjects. Andrology 2013; 1:838-844. doi: 10.1111/j.2047-2927.2013.00138.x.

[8] Skowronek F, Casanova G, Alciaturi J, Capurro A, Cantu L, Montes JM, et al. DNA sperm damage correlates with nuclear ultrastructural sperm defects in teratozoospermic men. Andrologia 2012; 44:59-65. doi: 10.1111/j.1439-0272.2010.01106.x.

[9] Candela L, Boeri L, Capogrosso P, Cazzaniga W, Pozzi E, Belladelli F, et al. Correlation among isolated teratozoospermia, sperm DNA fragmentation and markers of systemic inflammation in primary infertile men. PLoS One 2021; 16(6):e0251608. doi: 10.1371/journal.pone.0251608.

[10] Szczygiel M, Kurpisz M. Teratozoospermia and its effect on male fertility potential. Andrologia 1999; 31:63-75.

[11] Hotaling JM, Smith JF, Rosen M, Muller CH, Walsh TJ. The relationship between isolated teratozoospermia and clinical pregnancy after in vitro fertilization with or without intracytoplasmic sperm injection: A systematic review and meta-analysis. FertH Steril 2011; 95(3):1141-1141. doi: 10.1016/j.fertnstert.2010.09.029.

[12] World Health Organization. WHO laboratory manual for the examination of human semen and sperm-cervical mucus interaction. 4th ed. Geneva: World Health Organization; 1999. [Online] Available from https://www.aab.org/images/WHO%204th%20manual.pdf [Accessed 7 March 2023].

[13] World Health Organization. WHO laboratory manual for the examination and processing of human semen. 6th ed. World Health Organization; 2021. [Online] Available from https://apps.who.int/iris/handle/10665/343208 [Accessed 7 March 2023].

[14] Fischer MA, Willis J, Zini A. Human sperm DNA integrity: Correlation with sperm cytoplasmic droplets. Urology 2003; 61:207-211. doi: 10.1016/s0090-4295(02)02098-8.

[15] Khalili MA, Aghaie-Maybodi F, Anvari M, Talebi AR. Sperm nuclear DNA in ejaculates of fertile and infertile men: Correlation with semen parameters. Urol J 2006; 3:154-159.

[16] Braham A, Ghedir H, Zidi I, Sallem A, Hajlaoui A, Ajina M, et al. Nuclear sperm quality in total polymorphic teratozoospermia and its impact on intracytoplasmic sperm injection outcome. Andrologia 2019; 51(5):e13252. doi: 10.1111/and.13252.

[17] Jakubik-Uljasz J, Gill K, Rosiak-Gill A, Piasecka M. Relationship between sperm morphology and sperm DNA dispersion. Transl Androl Urol 2020; 9:405-415. doi: 10.21037/tau.2020.01.31.

[18] Siddique RA, Mohanarao GJ, Kumar R, Kumar A, Malik PK, Kumar C, et al. Sperm abnormalities and DNA fragmentation vis-à-vis Mammalian male infertility-A review. WJAS 2011; 174-189.

[19] Blom E. The ultrastructure of some characteristic sperm defects and a proposal for a new classification of the bull spermiogram. Nord Vet Med 1973; 25:383-391.

[20] Enciso M, Cisale H, Johnston SD, Sarasa J, Fernández JL, Gosálvez J. Major morphological sperm abnormalities in the bull are related to sperm DNA damage. Theriogenology 2011;76;23-32. doi: 10.1016/j.theriogenology.2010.12.034.

[21] Sailer BL, Jost LK, Evenson DP. Bull sperm head morphometry related to abnormal chromatin structure and fertility. Cytometry 1996; 24(2):167-173.

[22] Muriel L, Goyanes V, Segrelles E, Gosálvez J, Álvarez JG, Fernández JL. Increased aneuploidy rate in sperm with fragmented DNA as determined by the sperm chromatin dispersion (SCD) test and FISH analysis. J Androl 2007; 28:38-49. doi: 10.2164/jandrol.106.000067.

[23] Rives N, Saint Clair A, Mazurier S, Sibert L, Simeon N, Joly G, et al. Relationship between clinical phenotype, semen parameters and aneuploidy frequency in sperm nuclei of 50 infertile males. Hum Genet 1999; 105:266-272. doi: 10.1007/s004390051100.

[24] Lewis-Jones I, Aziz N, Seshadri S, Douglas A, Howard P. Sperm chromosomal abnormalities are linked to sperm morphologic deformities. Fertil Steril 2003; 79:212-215. doi: 10.1016/s0015-0282(02)04411-4.

[25] Calogero AE, De Palma A, Grazioso C, Barone N, Romeo R, Rappazzo G, et al. Aneuploidy rate in spermatozoa of selected men with abnormal semen parameters. Hum Reprod 2001; 16:1172-1179. doi: 10.1093/humrep/16.6.1172.

[26] Kalthur G, Adiga SK, Upadhya D, Rao S, Kumar P. Effect of cryopreservation on sperm DNA integrity in patients with teratospermia. Fertil Steril 2008; 89:1723-1727. doi: 10.1016/j.fertnstert.2007.06.087.

[27] Gorczyca W, Traganos F, Jesionowska H, Darzynkiewicz Z. Presence of DNA strand breaks and increased sensitivity of DNA in situ to denaturation in abnormal human sperm cells: Analogy to apoptosis of somatic cells. Exp Cell Res 1993; 207:202-205. doi: 10.1006/excr.1993.1182.