Evaluation of the Effects of Semen Incubation With ANDROSITOL®DGN on Sperm Motility and Mitochondrial Membrane Potential

Study Overview
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Brief Summary
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Mitochondria is the cellular organelle responsible for the production of the energy necessary to fuel sperm motility. It has been demonstrated that mitochondrial efficiency is correlated to the fertilizing capacity of the spermatozoon and to the production of high quality embryos. Mitochondria efficiency is measured in the laboratory setting by evaluating the mitochondrial membrane potential. Myo-inositol is the most represented stereoisomer of the family of inositols and is the only one physiologically concentrated within the seminal plasma. It is essential for sperm maturation and motility and its deficiency is also associated to a reduced sperm count. Myo-inositol promotes motility and allows recovering a higher number of sperm cells after swim-up, both in normospermic patients and in patients with altered seminal parameters. Scientific studies have shown that semen samples treated in vitro with ANDROSITOL®DGN, show an improvement in mitochondrial efficiency that results in an increase in spermatozoa progressive motility. Based on the percentage increase in the progressive motility showed by the spermatozoa after incubation with ANDROSITOL®DGN (ANDROSITOL®TEST), it is possible to subdivide the semen samples into three categories: low, medium, and high responders. The aim of the study is to evaluate whether the in vitro response of spermatozoa to ANDROSITOL®TEST correlates with the in vivo improvement of seminal parameters after oral treatment with antioxidants and myo-inositol.

Detailed Description
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Mitochondria is the cellular organelle responsible for the production of the energy necessary to fuel sperm motility. It has been demonstrated that mitochondrial efficiency is correlated to the fertilizing capacity of the spermatozoon and to the production of high quality embryos. Mitochondria efficiency is measured in the laboratory setting by evaluating the mitochondrial membrane potential. Myo-inositol is the most represented stereoisomer of the family of inositols and is the only one physiologically concentrated within the seminal plasma. It is essential for sperm maturation and motility and its deficiency is also associated to a reduced sperm count. Myo-inositol promotes motility and allows recovering a higher number of sperm cells after swim-up, both in normospermic patients and in patients with altered seminal parameters. Scientific studies have shown that semen samples, both pathological and normal, treated in vitro with ANDROSITOL®DGN - a concentrate solution (66X) containing 133 mg/ml of myo-inositol - show an improvement in mitochondrial efficiency that results in an increase in spermatozoa progressive motility. Based on the percentage increase in the progressive motility showed by the spermatozoa after incubation with ANDROSITOL®DGN (ANDROSITOL®TEST), it is possible to subdivide the semen samples into three categories: low, medium, and high responders. High responders have worst mitochondrial function and lower fertilizing capacity, and could represent the category of patients most benefiting from supplementary oral therapy with antioxidants and myo-inositol. The aim of our study is to evaluate whether the in vitro response of spermatozoa to ANDROSITOL®TEST correlates with the in vivo improvement of seminal parameters after oral treatment with antioxidants and myo-inositol. To do this, the investigators will enroll at least 13 patients for each category (low, medium, and high responder at ANDROSITOL®TEST) and they will re-evaluate conventional seminal parameters, mitochondrial function, and response to ANDROSITOL®TEST after three months of oral supplementation with ANDROSITOL® (dietary supplement of myo-inositol, vitamin E, L-carnitine, L-arginine, folic acid and selenium). The investigators hypothesize that, following supplementation, high-responder patients will exhibit the best improvement in seminal parameters, in particular in sperm motility. Furthermore, if the mitochondrial function is fully restored, they should respond less to the ANDROSITOL®TEST and could be reclassified as low responders.

Official Title
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Evaluation of the Effects of Semen Incubation With ANDROSITOL®DGN on Sperm Motility and Mitochondrial Membrane Potential Before and After Oral Supplementation With Antioxidants and Myo-inositol

Conditions
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Asthenozoospermia, Mitochondrial Damage

Intervention / Treatment
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* Diagnostic Test: ANDROSITOL®TEST


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: / Exclusion Criteria: - Absolute asthenozoospermia Leukocytospermia Positive semen culture and/or urethral swab Human Papilloma Virus (HPV) DNA in semen History of cryptorchidism 3rd degree varicocele Markedly reduced testicular volume Decompensated diabetes mellitus and other systemic diseases leading to oxidative stress (e.g. chronic renal failure, liver failure) Altered concentrations of the following hormones: luteinizing hormone (LH), follicle stimulating hormone (FSH), total testosterone, prolactin, 17β-estradiol Alcohol and drug abuse Heavy cigarette smoke (≥10 cigarettes/day) Body Mass Index (BMI) >35 kg/m2

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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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Diagnostic
Intervention Model: Single Group Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: ANDROSITOL®TEST<br>At least 45 patients (13 for each category: low, medium, and high responders, + 15% of hypothetical drop-outs) | Diagnostic Test: ANDROSITOL®TEST<br>* Sperm incubation with ANDROSITOL®DGN and evaluation of sperm motility and mithocondrial membrane potential<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Response to the ANDROSI-TEST | Number of poor and high responders to ANDROSI-TEST | T0 and T1 (three months) | 
| Sperm parameters | Percentage of total and progressive sperm motility and percentage of spermatozoa with high or low mitochondrial membrane potential | T0 and T1 (three months) | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Other Sperm parameters (1) | Sperm concentration (mil/ml) and Sperm total count (mil/ejaculate) | T0 and T1 (three months) | 
| Other Sperm parameters (2) | Percentage of spermatozoa with normal morphology | T0 and T1 (three months) | 
| Effects after therapy (1) | Re-evaluation of number of poor and high responders to ANDROSI-TEST 3 months after the discontinuation of Andrositol intake | T1 (three months) and T2 (six months - three months after supplementation withdrawal) | 
| Effects after therapy (2) | Re-evaluation of percentage of total and progressive sperm motility and percentage of spermatozoa with high or low mitochondrial membrane potential 3 months after the discontinuation of Andrositol intake | T1 (three months) and T2 (six months - three months after supplementation withdrawal) | 
| Effects after therapy (3) | Re-evaluation of sperm concentration (mil/ml) and Sperm total count (mil/ejaculate) 3 months after the discontinuation of Andrositol intake | T1 (three months) and T2 (six months - three months after supplementation withdrawal) | 
| Effects after therapy (4) | Re-evaluation of Percentage of spermatozoa with normal morphology 3 months after the discontinuation of Andrositol intake | T1 (three months) and T2 (six months - three months after supplementation withdrawal) | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Male infertility, Sperm parameters, Mitochondrial membrane potential