The advancement of reproductive medicine is somewhat slower than it is in other branches of medicine. Conventionally, it has been defined that it is the question of infertility if there is no pregnancy within 12 months of matrimony (or common-law marriage) with regular sexual intercourse and no birth control pills.
It is well known that men and women are the cause of infertile marriage in 45% of cases, respectively, and both of them in 10% of cases. Spermatogram analysis, in regular laboratory practice includes: sperm volume, sperm viscosity, pH value, normal morphologic features, spermatozoon count, the number of motile spermatozoa and rate of their motility. Normal spermatogram implies that over 50% of spermatozoa are motile.
Management of the existing anatomic malformations, hormonal disorders and associated infectious agents are standard procedures in reproductive medicine related to male. There is a great need for better and new approaches to this problem.
The rationale for the study “The effects of hyperbaric oxygen treatment on vigility of spermatozoids: preliminary report” is the fact that oxygen partial pressure (pPO2 ) is not measured either within regular laboratory tests or in specialized institutions. The investigators A Mitrović, P Brkić, T Jovanović consider that its value is crucial for managing the problem described in their study.
If the vigilance and motility of spermatozoa are mitochondria, energy-related, then, under conditions of regular energetic status, normal fructose level, the oxygen deficit may actually be the cause of deficiency or insufficiency of energy necessary for motility of male germ cells. Likewise, it may be the cause of accumulation of anaerobic products and altered intracellular pH value that again may lead to such reduction of motor activity that prevents their motility required for contact and penetration of ovular membrane.
In literature there are just a few organized investigations and appropriate results on possibilities of HBO in the management of male infertility and that fact has encouraged the investigators to proceed with their research. The aim of this study was to evaluate the effect of oxygen applied in hyperbaric conditions to the vigilance of spermatozoa.
Study population consisted of the group of 10 men, age between 28 and 35, with residence in Belgrade, Serbia. All of them were married for more than two years, without offspring. All of the patients have voluntarily come to the Hyperbaric medical Center after they have been diagnosed and unsuccessfully treated from oligospermia in other medical facilities. Absence of anatomical malformations of the reproductive system, normal morphologic features of spermatozoa, hormonal disorders and urinary infections were ratified in all of the patients.
A total of 10 specimens were obtained, one from each patient, by masturbation at the clinical site. Once obtained the specimens were kept at the room temperature and within 30 minutes they were examined in the laboratory. Only samples verified to be free from infectious agents, while sperm volume, pH value, fructose level and total count of spermatozoa were within normal limits, were included in future procedures.
The motility of spermatozoa in tested samples was not lower than 30% or it did not exceed the critical proportion of 50% (42.3±6.3). Each sperm specimen was divided in 3 samples and 1 ml of each was used for our experiment. Three groups of samples were formed, with respectively 10 samples in each group. Soon as the laboratory examinations were over, less than one hour after they were obtained, all three groups of samples were exposed to HBO conditions.
Samples were placed in specialized experimental hyperbaric chamber and were exposed to pure oxygen at the pressure of 2.5 absolute atmospheres (ATA) for 90 minutes, with 10 minutes periods of compression and decompression. The only difference among samples was the type of solution used for incubation during the treatment.
Group I samples were exposed to HBO conditions without any addition of medium. Each sample of group II was added 5 ml of solution that is generally used for separation of spermatozoa in artificial fertilization EBSS solution (in mM: 116.3 NaCl, 5.3 KCl, 26.1 NaHCO3 , 1.8 CaCl2 , 1.0 NaH2 PO4 , 0.8 MgSO4 , 5.5 D-glucose, and 0.03 phenol red). Tyrode’s solution (8 g of NaCl, 0.2 g of KCl, 0.2 g of CaCl2 , 0.1 g of MgCl2 , 0.05 g of NaH2 PO4 , 1 g of NaHCO3 , 1 g of d-glucose, and water to make 1000 ml) in volume of 5 ml was added to samples of group III. Solutions were purchased from Sigma-Aldrich S.r.l. Milan, Italy.
The basic evaluation of functional characteristics of sperm was carried out in time interval of 30 minutes after completion of HBO treatment, in the way it was performed with each sample prior to single HBO treatment. The research was approved by the Ethical Committee of the School of Medicine, University of Belgrade.
Providing sufficient amount of oxygen to the tissues and cells is crucial for maintaining theirs functional capabilities. The vigilance and motility of spermatozoa are mitochondria, energy-related activities. The presumption of the investigators was that by exposing the sperm samples to HBO conditions they will insure that, mitochondrial activity within spermatozoa can functions without any disturbance and reach the maximum of its capacities. Increase in motor activity that they registered confirmed that their presumption was correct.
The obtained results suggest that the acute exposure of sperm samples to high pressure of pure oxygen has a favorable impact on to functional capacity of spermatozoa in terms of better motility. In addition, the obtained results make this method of involving the patients into HBO treatment program recognized, and, accordingly it might help solving many aspects of the problem of male infertility.
Mitrović A, Brkić P, Jovanović T. Acta Physiol Hung. 2011 Mar;98(1):85-90.
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