Impact of Age and Fertility Status on the Consistency of Repeat Measurements of Sperm DNA Damage: A Single-center, Prospective, Dual Visit Study



      To examine the relationship between abnormal routine semen parameters and DNA damage associated with advanced paternal age and infertility by controlling for common biases contested in the current literature.

      Materials & Methods

      Prospective study using 151 semen samples collected from men aged 18-80 at 2 visits with 1-3 months intervals. Samples were collected from both infertile and general population controls. Conventional semen parameters were measured including volume, concentration and motility. Sperm DNA damage was measured using the %DNA Fragmentation Index (%DFI) and High DNA stainability (%HDS) using Sperm Chromatin Structure Assay (SCSA). Patients were then classified according to %DFI as normal (<18), intermediate (18-27), or high (>27).


      Significant correlation between all sperm parameters was seen between both visits regardless of age. DFI had the highest correlation between both visits (R2 = 0.77). Progressive motility, total motility and %DFI were significantly affected in men ≥50 years old when compared to men <35 and men 35-49 years old (P <.001). Forty-eight percent of men with intermediate %DFI changed category on their second visit, whereas men with high and low %DFI changed category in 15% and 9%, respectively.


      Sperm and SCSA parameters do not change significantly between 2 visits at 1-3 months intervals in the total population and after subgrouping. Men of advanced age have poorer sperm parameters and more DNA damage. Men with initially normal or elevated %DFI are unlikely to change DNA damage category. Older men are more likely to have sperm parameters and DNA damage vary on repeat semen analysis compared to younger men.
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        • World Health Organization
        • Programme of M, Child H, Family Planning U
        Infertility: A Tabulation of Available Data on Prevalence of Primary and Secondary Infertility.
        World Health Organization, Geneva1991
        • Tielemans E
        • Burdorf A
        • te Velde E
        • Weber R
        • van Kooij R
        • Heederik D.
        Sources of bias in studies among infertility clients.
        Am J Epidemiol. 2002; 156: 86-92
        • Hellstrom WJ
        • Overstreet JW
        • Sikka SC
        • et al.
        Semen and sperm reference ranges for men 45 years of age and older.
        J Androl. 2006; 27: 421-428
        • Cocuzza M
        • Athayde KS
        • Agarwal A
        • et al.
        Age-related increase of reactive oxygen species in neat semen in healthy fertile men.
        Urology. 2008; 71: 490-494
        • Henkel R
        • Maass G
        • Schuppe HC
        • Jung A
        • Schubert J
        • Schill WB.
        Molecular aspects of declining sperm motility in older men.
        Fertil Steril. 2005; 84: 1430-1437
        • Yatsenko AN
        • Turek PJ.
        Reproductive genetics and the aging male.
        J Assist Reprod Genet. 2018; 35: 933-941
        • Turek Pj
        Male Reproductive Physiology Campbell Walsh Wein Urology.
        12 ed. Elsevier, 2020: 1402
        • Evenson DP
        • Jost LK
        • Marshall D
        • et al.
        Utility of the sperm chromatin structure assay as a diagnostic and prognostic tool in the human fertility clinic.
        Hum Reprod. 1999; 14: 1039-1049
        • Spanò M
        • Bonde JP
        • Hjøllund HI
        • Kolstad HA
        • Cordelli E
        • Leter G.
        Sperm chromatin damage impairs human fertility. The Danish First Pregnancy Planner Study Team.
        Fertil Steril. 2000; 73: 43-50
        • Gunes S
        • Hekim GN
        • Arslan MA
        • Asci R.
        Effects of aging on the male reproductive system.
        J Assist Reprod Genet. 2016; 33: 441-454
        • Pino V
        • Sanz A
        • Valdés N
        • Crosby J
        • Mackenna A.
        The effects of aging on semen parameters and sperm DNA fragmentation.
        JBRA Assist Reprod. 2020; 24: 82-86
        • Varshini J
        • Srinag BS
        • Kalthur G
        • et al.
        Poor sperm quality and advancing age are associated with increased sperm DNA damage in infertile men.
        Andrologia. 2012; 44: 642-649
        • Petersen CG
        • Mauri AL
        • Vagnini LD
        • et al.
        The effects of male age on sperm DNA damage: an evaluation of 2,178 semen samples.
        JBRA Assist Reprod. 2018; 22: 323-330
        • Erenpreiss J
        • Bungum M
        • Spano M
        • Elzanaty S
        • Orbidans J
        • Giwercman A.
        Intra-individual variation in sperm chromatin structure assay parameters in men from infertile couples: clinical implications.
        Hum Reprod. 2006; 21: 2061-2064
        • Oleszczuk K
        • Giwercman A
        • Bungum M.
        Intra-individual variation of the sperm chromatin structure assay DNA fragmentation index in men from infertile couples.
        Hum Reprod. 2011; 26: 3244-3248
        • Agarwal A
        • Majzoub A
        • Esteves SC
        • Ko E
        • Ramasamy R
        • Zini A.
        Clinical utility of sperm DNA fragmentation testing: practice recommendations based on clinical scenarios.
        Transl Androl Urol. 2016; 5: 935-950
        • World Health O
        WHO Laboratory Manual for the Examination and Processing of Human Semen.
        5th ed. World Health Organization, Geneva2010
        • Komori K
        • Tsujimura A
        • Ishijima S
        • et al.
        Comparative study of Sperm Motility Analysis System and conventional microscopic semen analysis.
        Reprod Med Biol. 2006; 5: 195-200
        • Evenson DP
        • Djira G
        • Kasperson K
        • Christianson J.
        Relationships between the age of 25,445 men attending infertility clinics and sperm chromatin structure assay (SCSA) defined sperm DNA and chromatin integrity.
        Fertil Steril. 2020; 114: 311-320
        • Bungum M
        • Humaidan P
        • Spano M
        • Jepson K
        • Bungum L
        • Giwercman A.
        The predictive value of sperm chromatin structure assay (SCSA) parameters for the outcome of intrauterine insemination, IVF and ICSI.
        Hum Reprod. 2004; 19: 1401-1408
        • Sauer MV.
        Reproduction at an advanced maternal age and maternal health.
        Fertil Steril. 2015; 103: 1136-1143
        • Klipstein S
        • Regan M
        • Ryley DA
        • Goldman MB
        • Alper MM
        • Reindollar RH.
        One last chance for pregnancy: a review of 2,705 in vitro fertilization cycles initiated in women age 40 years and above.
        Fertil Steril. 2005; 84: 435-445
        • Gonzalez DC
        • Ory J
        • Blachman-Braun R
        • Nackeeran S
        • Best JC
        • Ramasamy R.
        Advanced paternal age and sperm DNA fragmentation: a systematic review.
        World J Mens Health. 2021; 40.1: 104
        • Borini A
        • Tarozzi N
        • Bizzaro D
        • et al.
        Sperm DNA fragmentation: paternal effect on early post-implantation embryo development in ART.
        Hum Reprod. 2006; 21: 2876-2881
        • Zini A
        • Boman JM
        • Belzile E
        • Ciampi A.
        Sperm DNA damage is associated with an increased risk of pregnancy loss after IVF and ICSI: systematic review and meta-analysis.
        Hum Reprod. 2008; 23: 2663-2668
        • Sun JG
        • Jurisicova A
        • Casper RF.
        Detection of deoxyribonucleic acid fragmentation in human sperm: correlation with fertilization in vitro.
        Biol Reprod. 1997; 56: 602-607
        • Xie P
        • Keating D
        • Parrella A
        • et al.
        Sperm genomic integrity by TUNEL varies throughout the male genital tract.
        J Urol. 2020; 203: 802-808
        • Stone BA
        • Alex A
        • Werlin LB
        • Marrs RP
        Age thresholds for changes in semen parameters in men.
        Fertil Steril. 2013; 100: 952-958
        • Verón GL
        • Tissera AD
        • Bello R
        • et al.
        Impact of age, clinical conditions, and lifestyle on routine semen parameters and sperm kinematics.
        Fertil Steril. 2018; 110: 68-75
        • Schlegel PN
        • Sigman M
        • Collura B
        • et al.
        Diagnosis and treatment of infertility in men: AUA/ASRM guideline part I.
        J Urol. 2021; 205: 36-43