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Infertility is a big problem that many couples worldwide have to face. In fact, it is now affecting nearly 15% of couples that, according to the World Health Organization (WHO), (1) fail to achieve a clinical pregnancy after 12 months or more of regular unprotected sexual intercourse. A population based-study carried out by Agarwal et al to estimate male infertility distribution around the world, showed that the male factor distribution in infertility was between 20% and 70% and the proportion of infertile men was within the 2.5%-12% range (2). The traditional way of diagnosing male infertility is by determining the classical semen parameters, as described in laboratory guidelines defined by the WHO (1). After semen analysis, normal semen parameters provide no firm conclusions to clinicians either about the fertility status of a patient or the outcomes of any infertility treatment. Various studies have demonstrated that abnormalities during the sperm chromatin organization may lead to male infertility (3-5) and may later influence the fertilization, the embryo quality and its development (6,7). This means that sperm DNA analysis with the standard semen analysis may help to reveal any hidden sperm DNA abnormality in infertile men with idiopathic infertility. In the light of the increasing body of evidence for DNA integrity and its importance for the ART outcome, many methods have been developed, within the last decade, to reveal any changes, such as fragmentation and the protamination, in the sperm chromatin status and maturity. But the use of such techniques as complementary biomarkers beside semen analysis is still controversial. Environmental and lifestyle factors, like nutrition, drinking alcohol, physical activity and tobacco smoke, play an important role in the aggravation of the idiopathic male infertility problem. In fact, exposure to external toxicants leads to different alterations during the various phases (mitotic, meiotic and post-meiotic) of spermatogenesis (8). As a matter of fact, current debate about these issues seems to confirm that the male fertility impairment is increasing and that this might indeed be associated with environmental factors and life-styles. However, in this study, we have focused on tobacco smoke and its effect on semen parameters, sperm DNA quality and clinical outcomes after intracytoplasmic sperm injection (ICSI) therapy. Abstract Objectives: The present study focused on tobacco smoke and its effect on semen parameters, sperm DNA quality (compaction and fragmentation) and clinical outcomes after intracytoplasmic sperm injection (ICSI) therapy Materials and Methods: The semen samples were divided according to smoking status into the following 2 groups, 98 heavy-smokers (G1) and 43 non-smokers (G2). Semen was prepared and purified using the PureSperm gradients according to the WHO guidelines 2010. Protamine deficiency (CMA3 positivity) was assessed by chromomycin CMA3 staining and sperm DNA fragmentation (sDF) by TUNEL assay. Results: The mean concentration and the total motility were significantly higher in G2 in comparison to G1 (P = 0.014, and P = 0.026 respectively) and the results were similar for the mean percent of the progressive motility and normal morphology (P = 0.0001). CMA3+ and sDF in G2 were significantly lower in comparison to G1 (20.35 ± 13.34% vs. 33.30 ± 22.33%, P = 0.001; 14.23 ± 13.07% vs. 26.68 ± 19.77%, P = 0.0001). Meanwhile, there were no significant differences in the ICSI outcomes, except for the pregnancy rate, which was significantly higher in G2 than in G1 (0.60 ± 0.49% vs. 0.38 ± 0.48%; P = 0.013). In G1, CMA3+ correlated negatively with sperm concentration (r = -0.233, P = 0.021) but positively with sDF (r = 0.484, P = 0.0001). In G2, sDF correlated negatively with progressive motility and morphologically normal spermatozoa (r = -0.304, p=0.047; r = -0.361, P = 0.017 respectively). Conclusions: The findings of this study revealed that tobacco smoking altered sperm parameters and later affected the pregnancy results in ICSI therapy. CMA3 and TUNEL tests are therefore useful as a supplementary test before any ART treatment to ensure a good prognosis.

Paternal Smoking in Relation to Sperm Quality and intracytoplasmic Sperm Injection Outcomes