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The comprehensive assessment of the male infertility factor requires the performance of a thorough laboratory examination of the male sperm. Apart from the conventional semen parameters, included in the basic semen analysis (spermiogram), modern scientific research provides a series of laboratory tests, which focus on the assessment of the functionality of spermatozoa and their potential insemination capability.
Such selected laboratory tests enable a more thorough assessment of the functional condition of the male sperm and contribute to:
The sperm function tests, regardless of the conventional spermatological profile, are an additional diagnostic tool enabling clinical doctors to provide better guidance to sub-fertile couples. There is no single laboratory test that may define, with full certainty,if a sperm is fertile or infertile. It is the combination of laboratory analyzes that indicates the fertility potential of the examined sperm.
The G. Lyberopoulos Spermatology Laboratory applies the principles of quality control provided by the NATIONAL ACCREDITATION SYSTEM and reputed international entities such as the World Health Organisation and the European Society of Human Reproduction and Embryology ESHREI. It participates in an External Quality Control System under the auspices of the Andrological Laboratory of the University of Karolinska, Sweden.
As part of the diagnostic services provided, the Laboratory performs the following sperm function tests, which provide information on the spermatozoa potential fertilization ability.
The impact of the male DNA damage on the reproductive effort has been correlated with the difficulty of fertilization and/or the creation of poor quality embryos. The combined assessment:
A. Chromatin Integrity Test
Chromatin is the structural complex of DNA and proteins that constitute the genetic material of spermatozoon in the form of chromosomes. The presence of granular (immature) or hyperoxidised (hyperstabilised) chromatin indicates a disorder during the packaging of the genetic material in the nucleus of spermatozoa, which has been associated with negative effects on the fertilization potential.
B. DNA Fragmentation Test:
The DNA molecule is in the form of a coiled double helix that is structurally stabilized by lateral bonds. If the bonds brake, the helix becomes unstable. The structural distortion in the sperm DNA, known as ‘Fragmentation’, is a measurable feature of its functionality that is not reflected in any of the conventional spermatological parameters (motility, concentration, morphology etc.). Men with spermatological parameters within the reference values, may still have a high DNA Fragmentation Index.
According to extensive scientific research, the integrity of the sperm DNA molecule has been associated with the quality and/or viability of embryos, mainly during the early stages of division (4- to 8-cell and more). DNA Fragmentation Index of high value is correlated to the negative outcome of reproductive effort, regardless of the applied fertilization method (normal, IUI, IVF, ICSI), through:
The main factors responsible for the high levels of sperm DNA fragmentation are:
The assessment of the sperm DNA Fragmentation Index is mainly recommended in cases of:
In addition, it provides a complete evaluation of the male fertilization potential during infertility investigation. The test can be particularly useful:
while assessing sperm quality of men who have received strong cytotoxic treatment.
Oxidative stress stress is one of the main factors of infertility and results from the imbalance between free oxygen radicals ROS and antioxidants in sperm.
Oxygen free radicals are important for the normal function of spermatozoa as they contribute to the acrosome reaction, spermatozoa maturation and fertilizing ability, however high concentration of ROS leads to oxidative stress and spermatozoa are particularly sensitive to this as they lack a fully organized antioxidant system. High ROS levels have been linked to increased DNA fragmentation, reduced motility
of sperm, reduced fertilizing ability, increase in morphologically abnormal sperm, as well as reduced sperm-egg interaction.
Therefore, the measurement of the value of oxidative stress becomes necessary, in order for the examinee to receive, if necessary, the appropriate antioxidant
treatment
Sperm activation aims to isolate spermatozoa from the seminal fluid which includes extra leukocytes, epithelial cells, immotile/morphologically abnormal/immature spermatozoa and micro-organisms, with the ultimate aim of obtaining a spermatozoa population that is at its highest possible percentage alive , with propulsive motion and normal morphology.
Sperm activation can also be used as a diagnostic tool by the clinician to assess the performance of the sample after its activation and to be able to decide on the suitability of the sample for normal conception, for insemination or to decide whether it is suitable only for use in IVF.
Longevity describes how long sperm motility is maintained over time. In vitro analysis of sperm longevity can provide information regarding the potential fertilizing ability of sperm and help coordinate intercourse with ovulation.
The separation of immotile spermatozoa in a sample into live and dead is of crucial importance, especially in cases with a high percentage of immotile spermatozoa.
The control of the integrity and normal membrane function of the spermatozoa is evaluated through the Hypo-osmotic Swelling Test (HOS test).
Regarding the outcome of the reproductive effort, high values of the index constitute positive prognostic information. Conversely, in cases of presence of low levels of the indicator, the probability of in vivo conception is significantly reduced. Even when applying classic IVF, although fertilization rates do not seem to be affected, the chance of pregnancy remains low. In such cases, assisted reproduction shows better results with the ICSI method.
One of the important functional properties of sperm, is cervical mucus penetration, which directly depends on the spermatozoa motility and/or morphology, as well as certain kinematic parameters. The results of penetration tests provide:
Cervical mucus penetration failure a cause of subfertility that may result from:
The relevant laboratory tests performed at the Spermatology Laboratory include:
The ad hoc suitable laboratory tests regarding the interaction of sperm and cervical mucus are indicated in cooperation with the attending doctor.
The presence of anti-sperm antibodies is related to infertility in 9-13% of couples unable to conceive. Anti-sperm antibodies may be present in either one or both partners. A complete immunological examination of infertile couples includes screening for the detection of the two most important anti-sperm immunoglobulins, IgG and IgA, in spermatozoa or sperm fluid and blood serum of men, as well as in cervical mucus or blood serum of women.
Thedegree the anti-spermantibodies affect the fertilization processdependson:
The tests applied in the Spermatology Laboratory include:
Titration of anti-sperm antibodies that cause agglutinations (Friberg test)
The success of anassisted reproductive attempt strongly dependsonthe selection of the best spermatozoa in terms of quality. This step plays an important role, mainly when applying the ICSI method, where individual spermatozoa are selected and injected directly to the eggs.
According to a variety of modern studies, mature and functional spermatozoa that are capable of being attached to the zona pellucidaof the egg, also have the ability to bind to special hyaluronic acid polymers. Therefore, the sperm-hyaluronan binding test provides clinical doctors with the necessary information for the selection of the most suitable assisted reproduction method (IUI, IVF, ICSI) in view of optimizing the success rate.
It is a laboratory test that identifies the percentage of the best quality spermatozoa in the semen sample, which are then isolatedand used in assisted reproduction methods, with:
10% of infertile men are diagnosed with azoospermia. Azoopermia is defined as the complete absence of spermatozoa in the seminal fluid.
This clinical condition requires a special investigation and therapeutic strategy.
Azoospermia is divided into obstructive and non-obstructive. This distinction between the two categories is decisive both for the man himself and for the clinician for the further management of each case.
The use of biomarkers in recent years presents advantages in terms of diagnosis and the selection of appropriate treatment.
Biomarkers are molecules in a biological system with prognostic value. Differences in their concentration are associated with disturbances in the orderly functioning of this system.
The use of biomarkers in the investigation of male infertility is an innovation.
The ECM1 biomarker is a protein expressed in high concentration in epididymal tissue. It contributes to the maintenance of tissue integrity through interaction with a variety of structural and extracellular proteins. The measurement of the levels of the biomarker ECM1, which can now be performed in an andrological laboratory, helps to distinguish obstructive from non-obstructive azoospermia.
Since the development of surgical techniques for obtaining spermatozoa from the male genital system makes it possible to obtain offspring, in azoospermic men the examination of ECM1 levels can be an important tool for the detailed investigation and proper management of these cases.
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