Abstract
The aim of this study was to determine the relationship between the size-number of nuclear vacuoles and DNA damage in spermatozoa from patients with different sperm morphology index. Prospective-comparative study. Forty four men who undertwent ART (2011-2012) were selectec for this study. Accordingly to the WHO manual for semen examination (1999), the inclusion criteria were: sperm concentration ≥20 million/mL, progressive motility ≥30% and strict sperm morphology (Kruger) ≥14% (group 1), 5-13 (group 2) and ≤4% (group 3). After gradient, motile sperm were placed in PVP and selected considering by the number and size of nuclear vacuoles under high magnification (7800X). It was considered the following patterns: A) No vacuoles, B)1 vacuole, size=less than 25% of the nuclear surface, C)2 vacuoles less than 25%, D)1 vacuole greater than 25-50%, E)several vacuoles less than 25% and F)several vacuoles greater than 25%. Selected spermatozoa (by ICSI pipette) were assessed for DNA fragmentation (TUNEL) by immunofluorescence. Statistical analysis was performed by ANOVA. The mean age was: 35±4.5(group 1), 36±3.5 (group 2) and 36±3.0 (group 3). Progressive sperm motility was 59±22 (1), 55±18 (2), 60±18 (3). The average of Kruger was: 16.2±2 (1), 7.5±2.5 (2) and 1.2±1.4 (3). A total of 818 spermatozoa were selected and evaluated for TUNEL. The comparison between sperm populations are shown in table 1.Tabled 1GroupsABCDEFGroup 13.6±2.46.1±4.26.7±3.912.1±3.6ap<0.05.7.3±3.710.8±4.9ap<0.05.Group 25.8±2.26.2±2.97.5±3.113.2±3.4ap<0.05.9.6±2.212.8±3.6ap<0.05.Group 316.8±5.818.2±5.717.4±5.921.4±3.620.4±4.620.6±5.9a p<0.05. Open table in a new tab Sperm DNA fragmentation levels signifantly increases just in spermatozoa with large (D) or several nuclear vacuoles (F) in patients with Kruger between 5-13% or ≥14%, but not in patients with Kruger index ≤4%. This data may be relevant during patients and sperm selection for IMSI. Finally, sperm nuclear vacuoles are not always related with DNA fragmentation.
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