Administration of Antioxidants to Infertile Men and Sperm Quality

Study Overview
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Brief Summary
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The purpose of this randomized clinical trial is to assess the effect of oral antioxidant administration to infertile men, by evaluating semen variables, sperm DFI and levels of ROS. Oral antioxidants or placebo will be given for 3 consecutive months. The study will recruit infertile men, who have one previous abnormal spermiogram, with at least one pathological variable (concentration, motility, morphology), according to WHO 2010 criteria. Participants will be recruited in the outpatient clinic of the Unit of Human Reproduction and of the Unit of Reproductive Endocrinology at the 1st Ob/Gyn Dept.

Detailed Description
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Infertility affects up to 15% of couples trying to conceive with the male factor contributing in about 50% of cases. Oxidative stress (OS) has been found as one of the many factors causing male infertility, by inducing sperm damage. OS causes cell damage due to the raised production of reactive oxygen species (ROS), that overcome the antioxidant mechanisms of the human body. ROS are oxidizing agents that are produced as a by-product of oxygen metabolism, having at least one free electron. Due to this free electrone in their outer layer, the form very active or unstable substances. While small amounts of ROS are necessary for the proper function of sperm, increased amounts have a negative effect on sperm quality and can harm its fertility potential, thus inducing male infertility. Spermatozoa were the first cells found to be sensitive to OS, as they lack the necessary repair cytoplasmic enzyme systems, thus being unable to repair damage. In addition, their cytoplasmic membranes are rich in polyunsaturated fatty acids (PUFAs), making them highly susceptible to oxygen-induced damage, such as lipid peroxidation (LPO). Rapid loss of intracellular adenosine triphosphate (ATP) by LPO, is responsible for axial damage, morphological damage to the sperm neck and reduced sperm motility; these events contribute to reduced sperm motility. OS has also been associated with reduced fertilization, delayed intrauterine growth, miscarriages, birth defects (including autism) and childhood cancer. ROS found in semen come from a variety of endogenous and exogenous factors. Human sperm includes mature and immature sperm cells, leukocytes, and white blood cells. The leukocytes (mainly neutrophils and macrophages) and the immature spermatozoa are considered the major endogenous sources of ROS, whereas lifestyle, such as smoking, excessive alcohol consumption and other environmental factors such as radiation and toxins may contribute to the production of exogenous ROS. As oxidative stress (OS) results from the imbalance of ROS overproduction and the reduced capacity of sperm antioxidant systems, many studies have aimed to improve sperm quality by administering antioxidant therapeutic regimens. In general, antioxidant therapy involves the administration of oral antioxidants, and the in vitro addition of antioxidants to culture media that are used for sample preparation in assisted reproduction techniques (ART). Many different antioxidants combinations have been studied. More and more studies have shown the beneficial effect of antioxidants on reducing sperm fragmentation and improving sperm quality. Agarwal & Sekhon's study showed a positive correlation between antioxidant therapy and various semen parameters. However, no firm conclusion regarding the beneficial effect of oral antioxidants can be made, as the majority of available studies had small sample size, used different antioxidant combinations, and the techniques that were used for the detection of ROS and DNA fragmentation index (DFI) were not standardized. AIM OF THE STUDY The purpose of this randomized clinical trial is to assess the effect of oral antioxidant administration to infertile men, by evaluating semen variables, sperm DFI and levels of ROS.Oral antioxidants or placebo will be given for 3 consecutive months. This study will take place at the Unit of Reproductive Endocrinology at the 1st Ob/Gyn Dept, Medical School, Aristotle University of Thessaloniki in collaboration with the private andrology diagnostic center of Mr Th. Zeginiadou. PATIENTS The study will recruit infertile men, who have one previous abnormal spermiogram, with at least one pathological variable (concentration, motility, morphology), according to WHO 2010 criteria. OUTCOMES Main Outcome: WHO Sperm analysis variables: concentration, motility, morphology. Secondary outcomes: Concentration of ROS in sperm and sperm DFI STUDY DESIGN Type of study: Randomized clinical trial The study will recruit infertile men, who have one previous abnormal spermiogram, with at least one pathological variable (concentration, motility, morphology), according to WHO 2010 criteria. Participants will be recruited in the outpatient clinic of the Unit of Human Reproduction and of the Unit of Reproductive Endocrinology at the 1st Ob/Gyn Dept. Selected participants will sign the consent form and will be asked to sample sperm at the private andrology diagnostic center of Mr Th. Zeginiadou. The sperm samples will be evaluated regarding the concentration, motility, morphology and vitality of spermatozoa, as well as the concentration of ROS, and DFI. Oral antioxidants or placebo will be given for 3 consecutive months. At the end of the 3 months period participants will be asked to re-sample their sperm in order to evaluate the same variables. Sample size calculation Estimated sample size for two-sample means test, assuming and common standard deviation of 15 and a mean motility of 20 in the control group and 30 in the antioxidant group results in calculated sample size of 74. To allow for dropouts 80 patients will be recruited.

Official Title
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Does Antioxidant Administration Improve Sperm Quality in Infertile Men

Conditions
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Male Infertility

Intervention / Treatment
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* Other: Spermotrend


Participation Criteria
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Eligibility Criteria
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Inclusion criteria Men, 18-50 years old Infertility defined as follows: Failure to obtain a pregnancy after at least twelve (12) months of regular sexual intercourse without the use of contraceptives or six (6) months if the woman is> 35 years old AND At least one previous abnormal spermiogram, with at least one pathological parameter (concentration, motility, morphology), according to the WHO 2010 criteria. No treatment for infertility in the last three (3) months Normal hormone profile (TSH, FSH, LH, total testosterone, prolactin) Negative culture for mycoplasma or ureaplasma Physiological scrotal ultrasound Exclusion criteria Genetic cause of infertility History of cryptorchidism History of orchectomy History of testicular cancer History of severe heart, liver or kidney disease History of endocrine disease (primary or secondary hypogonadism, hyperprolactinemia, thyroid, pituitary or adrenal disease) History of systemic disease or treatment in the last three (3) months BMI > 30 kg/m2 Participation in another study and the possibility of the patient not being available for follow-up

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 50 Years

Sexes Eligible for Study
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Male

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Allocation: Randomized
Intervention Model: Parallel Assignment
Interventional Model Description: quadruple-blind study
Masking: Quadruple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Intervention Group<br> | Other: Spermotrend<br>* oral administration of antioxidants for three months<br>|
| Placebo Comparator: Control Group<br> | Other: Spermotrend<br>* oral administration of antioxidants for three months<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Sperm parameters | Sperm motility(Sperm motility was classified using a four-category scheme: A:rapid progressive, B:slow progressive, C:non-progressive, and D:immotile , %A+B, normal values >32% A+B) | immediately before treatment initiation | 
| Sperm parameters | Sperm motility(Sperm motility was classified using a four-category scheme: A:rapid progressive, B:slow progressive, C:non-progressive, and D:immotile , %A+B, normal values >32% A+B) | immediately after the end of a 3 month treatment | 
| Sperm parameters | Sperm concentration (ml, normal values > 1.5 ml) | immediately before treatment initiation | 
| Sperm parameters | Sperm concentration (ml, normal values > 1.5 ml) | immediately after the end of a 3 month treatment | 
| Sperm parameters | Sperm vitality (%, normal values > 58%) | immediately before treatment initiation | 
| Sperm parameters | Sperm vitality (%, normal values > 58%) | immediately after the end of a 3 month treatment | 
| Sperm parameters | Sperm morphology(Tygerberg Strict Criteria were used for the evaluation of human sperm morphology, normal form per 100 sperm, normal values >4%) | immediately before treatment initiation | 
| Sperm parameters | Sperm morphology(Tygerberg Strict Criteria were used for the evaluation of human sperm morphology, normal form per 100 sperm, normal values >4%) | immediately after the end of a 3 month treatment | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Reactive Oxygen Species (ROS) | ROS (8-hydroxy-2-deoxy-quanosine%, normal values < 3% on sperm) | immediately before treatment initiation | 
| Reactive Oxygen Species (ROS) | ROS (8-hydroxy-2-deoxy-quanosine%, normal values < 3% on sperm) | immediately after the end of a 3 month treatment | 
| DNA fragmentation Index (DFI) | DFI (%DFI: % sperm cells containing damaged DNA) | immediately before treatment initiation | 
| DNA fragmentation Index (DFI) | DFI (%DFI: % sperm cells containing damaged DNA) | immediately after the end of a 3 month treatment | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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sperm, infertility, asthenospermia, oligospermia, teratozoospermia, antioxidants