Center for Reproductive Medicine - Innovations in Male Infertility Research

INNOVATIONS IN MALE INFERTILITY RESEARCH

Cleveland Clinic's Center for Reproductive Medicine
1993 - 2008

Oxidative Stress and Male Infertility

A. Understanding oxidative stress

Oxidative stress (OS) is the imbalance between the production of reactive oxygen species (ROS) by the spermatozoa and leukocytes and the antioxidant capacity of the seminal plasma. The primary source of ROS production in infertile patients is the immature spermatozoa having residual cytoplasm and leukocytes (granulocytes). Oxidative stress can damage the DNA of the spermatozoa and prevent them from fertilizing an egg. It is involved in the pathophysiology of male infertility. A large number of comprehensive reviews have been published that establish the role of OS in infertility and the potential benefits of antioxidant supplementation both in vitro and in vivo on sperm quality.

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Sharma RK, Agarwal A. (1996):
Role of reactive oxygen species in male infertility. Review article.
Urology 48:835-850.
PDF File (osdoc40.pdf 135 Kb)
Agarwal A, Saleh RA (2002):
The role of oxidants in male infertility: rationale, significance, and treatment.
Urologic Clinics of North America: Male Infertility - 2002 (Editors: L.I. Lipshultz and C. Niederberger), W.B. Saunders Co, Philadelphia, Pa, Vol. 29, Issue 4, November 2002, pp. 817-827.
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Saleh RA, Agarwal A (2002):
Oxidative stress and male infertility: from research bench to clinical practice. Review article.
J Androl 23:737-752.
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Agarwal A, Saleh RA, Bedaiwy MA (2003):
Role of reactive oxygen species in the pathophysiology of human reproduction. Review article.
Fertil Steril 79:829-843.
PDF File (agradoc127.pdf 134 Kb)
Esfandiari N, Sharma RK, Saleh RA, Thomas AJ Jr, Agarwal A (2003):
Utility of the nitroblue tetrazolium (NBT) reduction test for assessment of reactive oxygen species production by seminal leukocytes and spermatozoa.
J Androl 24:862-870.
PDF File (agradoc139.pdf 450 Kb)
Agarwal A, Nallella KP, Allamaneni SR, Said TM (2004):
The role of antioxidants in treatment of male infertility: an overview of the literature.
Reprod Biomed Online 8:616-627.
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Agarwal A, Allamaneni SSR (2004):
Oxidants and antioxidants in human fertility. Review article.
MEFS Journal 9:187-197.
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Aziz N, Agarwal A, Lewis-Jones I, Sharma RK, Thomas AJ Jr (2004):
Novel associations between specific sperm morphological defects and leukocytospermia.
Fertil Steril 82:621-7.
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Agarwal A, Said TM (2005):
Oxidative stress, DNA damage and apoptosis in male infertility - a clinical approach. Review article.
BJU International 95:503-7.

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Agarwal A (2005):
Role of oxidative stress in male infertility and antioxidant supplementation. Business Briefing.
US Kidney and Urological Disease 122-122.
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Agarwal A, Prabakaran SA, Said TM (2005):
Strategies to prevent oxidative stress-induced injury to human spermatozoa.
Review article. J Androl 26:654-60.
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Agarwal A, Prabakaran SA (2005):
Mechanism, measurement, and prevention of oxidative stress in male reproductive physiology. Review article.
Indian J Exp Biol 43:963-74.
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Agarwal A, Prabakaran SA (2005):
Oxidative stress and antioxidants in male infertility - a difficult balance. Review article.
Iran J Reprod Med 3:1-8.
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Agarwal A, Prabakaran SA, Allamaneni SSR (2006):
What an andrologist/urologist should know about free radicals and why. Review article.
Urology 67:2-8.
PDF File (agradoc209.pdf 430 Kb)
Agarwal A, Prabakaran SA, Allamaneni SSR (2006):
Relationship between oxidative stress, varicocele and infertility: a meta-analysis. Review article.
Reprod BioMed Online 12: 630-633.
PDF File (agradoc211.pdf 103 Kb)
Agarwal A, Gupta S, Sikka S (2006):
The role of free radicals and antioxidants in reproduction. Review article.
Curr Opin Obstet Gynecol 18:325-332.
PDF File (agradoc234.pdf 617 Kb)
Agarwal A, Prabakaran SA, Sikka S (2007):
Clinical relevance of oxidative stress in patients with male factor infertility: evidence-based analysis. Review Article.
AUA Update 26: 1-12, Lesson 1, 2007.
PDF File (agradoc261.pdf 262 Kb)
Agarwal A, Makker, K, Sharma, RK (2007):
Clinical relevance of oxidative stress in male factor infertility: an update. Review article.
AJRI 59: 2-11.

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B. Sources of oxidative stress

Abnormal spermatozoa are not fully mature, and in the presence of white blood cells they can produce harmful, very unstable substances called free radicals or ROS. Free radicals such as superoxide anion (O2-·), hydrogen peroxide (H2O2), hydroxyl (OH-), and peroxyl radicals are involved in initiation and progression of oxidative damage to spermatozoa. When the level of these ROS exceeds the antioxidant capacity of the cell, oxidative stress will be induced, which negatively impacts male fertility potential. We have standardized laboratory techniques to measure reactive oxygen species in seminal ejaculates. ROS can be measured in unprocessed liquefied seminal ejaculate, wash and resuspended sperm and those prepared on a density gradient centrifugation. ROS can be measured by chemiluminescence assay using a luminometer.

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Agarwal A, Ikemoto I, Loughlin KR (1994):
Effect of sperm washing on reactive oxygen species level in semen.
Arch Androl 33:157-162.
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Agarwal A, Ikemoto I, Loughlin KR (1994):
Relationship of sperm parameters to levels of reactive oxygen species in semen specimens.
J Urol 152:107-110.
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Agarwal A, Ikemoto , Loughlin KR (1994):
Levels of reactive oxygen species before and after sperm preparation: comparison of swim-up and L4 filtration methods.
Arch Androl 32:169-174.
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Shekarriz M, Sharma RK, Thomas AJ, Jr, Agarwal A (1995):
Positive myeloperoxidase staining (Endtz Test) as an indicator of excessive reactive oxygen species formation in semen.
J Assist Reprod Genet 12:70-74.
PDF File (agradoc23.pdf 174 Kb)
Shekarriz, M, Dewire DM, Thomas AJ, Jr, Agarwal A (1995):
A method of human semen centrifugation to minimize the iatrogenic sperm injuries caused by reactive oxygen species.
Eur Urol 28:31-35.
PDF File (agradoc017.pdf 755 Kb)
Shekarriz M, Thomas AJ, Jr, Agarwal A. (1995):
Effect of time and sperm concentration on reactive oxygen species formation in human semen.
Arch Androl 34:69-75.
PDF File (agradoc018.pdf 1015 Kb)
Shekarriz M, Thomas AJ Jr, Agarwal A (1995):
Incidence and level of seminal reactive oxygen species in normal men.
Urology 45:103-107.
PDF File (agradoc35.pdf 524 Kb)
Padron OF, Brackett NL, Sharma RK, Lynne CM, Thomas AJ Jr, Agarwal A (1997):
Seminal reactive oxygen species, sperm motility and morphology in men with spinal cord injury.
Fertil Steril 67:1115-1120.
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Wang Y, Sharma RK, Agarwal A (1997):
Effect of cryopreservation and sperm concentration on lipid peroxidation in human semen.
Urology 50:409-413.
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Kobayashi H, Gil-Guzman E, Mahran, AM, Sharma RK, Nelson DR, Thomas, AJ Jr, Agarwal A (2001):
Quality control of reactive oxygen species measurement by luminol-dependent chemiluminescence assay.
J Androl 22:568-574.

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Gil-Guzman E, Ollero M, Lopez, MC, Sharma RK, Alvarez JG, Thomas AJ, Jr, Agarwal A (2001):
Differential production of reactive oxygen species by subsets of human spermatozoa at different stages of maturation.
Hum Reprod 16:1922-1930.
PDF File (agradoc98.pdf 216 Kb)
Ollero M, Gil-Guzman E, Lopez MC, Sharma RK, Agarwal A, Larson K, Evenson D, Thomas AJ Jr, Alvarez JG (2001):
Characterization of subsets of human spermatozoa at different stages of maturation: implications in the diagnosis and treatment of male infertility.
Hum Reprod 16:1912-1921.
PDF File (agradoc111.pdf 65 Kb)
Esfandiari N, Saleh RA, Blaut AP, Sharma RK, Nelson DR, Thomas AJ Jr, Falcone T, Agarwal A (2002):
Effects of temperature on sperm characteristics and reactive oxygen species production.
Int J Fertil & Women's Med 47:227-233.
PDF File (agradoc127.pdf 134 Kb)
Esfandiari N, Sharma RK, Saleh RA, Thomas AJ Jr, Agarwal A (2003):
Utility of the nitroblue tetrazolium (NBT) reduction test for assessment of reactive oxygen species production by seminal leukocytes and spermatozoa.
J Androl 24:862-870.
PDF File (agradoc122.pdf 47 Kb)
Said TM, Kattal N, Sharma RK, Sikka SC, Thomas AJ Jr, Mascha E, Agarwal A (2003):
Enhanced chemiluminescence assay versus colorimetric assay for measurement of the total antioxidant capacity of human seminal plasma.
J Androl 24:676-680.
PDF File (agradoc153.pdf 280 Kb)
Said TM, Agarwal A, Sharma RK, Mascha E, Sikka SC, Thomas AJ Jr (2004):
Human sperm superoxide anion generation and correlation with semen quality in patients with male infertility.
Fertil Steril 82:871-7.
PDF File (agradoc169.pdf 80 Kb)
Allamaneni SSR, Agarwal A, Nallella KP, Sharma RK, Thomas AJ Jr, Sikka SC (2005):
Characterization of oxidative stress status by evaluation of reactive oxygen species levels in whole semen and isolated spermatozoa.
Fertil Steril 83:800-3.
Athayde KS, Cocuzza M, Hallak,J, Agarwal A, Lucon AM, Srougi M (2007):
Development of normal reference values for reactive oxygen species and their correlation with leukocytes and semen parameters in a fertile population: a prospective controlled study.
J Androl 28:613-20.