Demonstration of the DNA Integrity in Semen Samples with Normospermia, Oligozoospermia and Azoospermia by Comet Method and Immunohistochemical Determination of EGF-R and TGF-alpha

Objective: The aim of this study was to compare the rate of sperm cells with DNA damage by the comet assay, and to determine EGF-R and TGF-alpha localization by using immunohistochemical methods in normospermic, oligozoospermic and azoospermic events. Methods: Semen samples were obtained from 30 men attending the Infertility Clinic of Baskent University Hospital. We used the comet assay to determine the DNA damage of sperm cells. Indirect immunohistochemical methods were used with EGF-R and TGF-alpha polyclonal antibodies. Results: To determine the DNA damage of sperm cells, the comet assay was used in normospermic, oligozoospermic and azoospermic events and a statistically significant difference was found among the treatment groups (p=0.038). The differences between normospermic and azoospermic events is greater than normospermic and oligozoospermic events (p=0.05). Furthermore, EGF-R and TGF-alpha immunoreaction on the sperm cell surface of normospermic and oligozoospermic events were stronger than azoospermic sperm cells. Reactivity of these growth factors was related with sperm morphology. Conclusion: In azoospermic semen samples, DNA damage was higher, and also the reactions for EGF-R and TGF-alpha were weaker when compared to normal semen samples.