[doi: 10.5505/2017ichc.PP-162]

Assessment of Sperm DNA İntegrity From Normozoospermic İnfertile Men Selected by Different Selection Methods

¹Department of Histology and Embryology, Abant Izzet Baysal University, Bolu, Turkey
²Department of Histology and Embryology, Kocaeli University, Kocaeli, Turkey

INTRODUCTION:
The male factor is responsible for about %50 of infertility cases and clinical parameters are not always enough for assessing the sperm quality of patients (1). The assessment of sperm DNA integrity was shown to be more significant than the standard semen analysis in determining the fertility potential. Density gradient and swim up methods are widely used classical methods for sperm selection at IVF treatment (2).
The microfluidic chip has been developed as a new sperm selection method which could be considered as a method of choice to achieve better results in IVF (3). The commonly used density gradient and swim up sperm selection methods require multiple centrifugation, washing or pipetting procedures that cause several sperm DNA damages. However, the microfluidic chip method does not require centrifugation and other common procedures to select good quality sperm (4). Although this microfluidic chip is an effective way to select good quality sperm, it is not clearly known whether it reduces sperm DNA fragmentation.
In this study density gradient and swim up which are classical selection techniques have been compared with microfluidic chip at the normozoospermic infertile patients.

MATERIALS-METHODS:

Semen samples from twenty normozoospermic infertile men were separated into four groups and semen samples sorted using by microfluidic chip, swim-up and density gradient methods. Sperm samples were measured according to the WHO guidelines and stained with Aniline Blue (figure 1), Toluidine Blue (figure 2) and TUNEL (figure 3) to determine sperm chromatin condensation and DNA fragmentation. Subsequently, semen parameters, the percentages of sperm DNA fragmentation and chromatin condensation defects were measured and analyzed by variance and bonferroni test. We used the software of SPSS version 21 for statistical analysis.

Result
When the methods are compared, we found significantly different (P<0.0001) at DNA integrity. It was seen that the chromatin condensation defects and DNA fragmentation in sperms were the lowest after microfluidic chip and the highest after density gradient method. The results show that the microfluidic chip method is a more effective and physiological method than the density gradient and swim up methods in the IUI treatment of normozoospermik infertile patients.

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figure 1

Aniline blue staining method, light blue sperm heads (black arrow) show normal chromatin condensation and dark blue sperm heads (red arrow) show abnormal chromatin condensation.

figure 2

Toluidine blue staining method, pale blue sperm heads (black arrows) show normal and dark blue or violet or purple sperm heads (red arrows) show abnormal chromatin condensation.

figure 3

TUNEL staining method, brown stained sperm heads show fragmented DNA and green colored sperm heads show non-fragmented DNA.