The effect of cotinine on spermatozoa quality and outcome of patients undergoing IVF/ICSI therapy

Abstract

OBJECTIVE: The objectives of this study were to determine the relationship between reactive oxygen species (ROS), total antioxidant (TAS) and Cotinine levels in seminal plasma and sperm parameters as well as their effect on IVF/ICSI outcome.DESIGN: Prospective study.MATERIALS AND METHODS: Semen concentration of ROS, TAS and sperm parameters concentration, motility, motility, vitality, membrane integrity (HOS-test), morphology, chromatin integrity (CMA3), DNA Fragmentation (Tunel-test) of 46 patients (11) heavy smokers and (35) non smokers were investigated. As semen liquefication and assessment, the semen was separated from seminal plasma using PureSperm gradient centrifugation. The seminal plasma on the top of the upper layer of the two fraction 45% and 90% of PureSperm layer was withdrawn after centrifugation for sperm preparation and kept frozen at - 80°C until the concentrations of ROS, TAS, and Cotinine were measured by colorimetric assay using commercially available kits. Cotinine concentration was measured with (ELISA) kit. The motile sperm which obtained after sperm preparation was used for injection (ICSI) or insemination of Oocytes (IVF).RESULTS: The sperm concentration, mean number of motile spermatozoa after sperm preparation, morphologically normal spermatozoa, and mean number of cleaved and transferred Oocytes and pregnancy rate were significantly higher in non-smoking patients group in comparison to smoking one (76.2±29.5mill/ml; 65.2±31.4%; 16.1±18.5%; 1.9±0.8%; 34%vs.50.7±39.2 mill/ml; 39.0±20.4%19.8±18.5%; 1.9±0.7% and 27.3% respectively). The other sperm parameters were similar in both groups. Cotinine concentration was significantly higher in smokers than non-smokers (144.3±82.2μg/dl vs.19.3±23.7μg/dl; p=0.001). Inverse correlations were found between Cotinine concentration and ejaculate volume, sperm concentration, motility, vitality, morphology membrane integrity, DNA fragmentation (TUNEL), TAS concentration and fertilization. Besides, a positive correlation was shown between Cotinine and ROS concentration. In the smoker group three patients become pregnant (27.3%) and 11 patients achieved pregnancy in the non-smoking group (34%; p=0.001).CONCLUSIONS: Cotinine concentration in seminal plasma has an adverse effect on seminal plasma anti oxidants, sperm parameters and ART outcome and increase the production of ROS in seminal plasma. Therefore, it is recommended to quit smoking in childless patients who are seeking ART therapy. OBJECTIVE: The objectives of this study were to determine the relationship between reactive oxygen species (ROS), total antioxidant (TAS) and Cotinine levels in seminal plasma and sperm parameters as well as their effect on IVF/ICSI outcome. DESIGN: Prospective study. MATERIALS AND METHODS: Semen concentration of ROS, TAS and sperm parameters concentration, motility, motility, vitality, membrane integrity (HOS-test), morphology, chromatin integrity (CMA3), DNA Fragmentation (Tunel-test) of 46 patients (11) heavy smokers and (35) non smokers were investigated. As semen liquefication and assessment, the semen was separated from seminal plasma using PureSperm gradient centrifugation. The seminal plasma on the top of the upper layer of the two fraction 45% and 90% of PureSperm layer was withdrawn after centrifugation for sperm preparation and kept frozen at - 80°C until the concentrations of ROS, TAS, and Cotinine were measured by colorimetric assay using commercially available kits. Cotinine concentration was measured with (ELISA) kit. The motile sperm which obtained after sperm preparation was used for injection (ICSI) or insemination of Oocytes (IVF). RESULTS: The sperm concentration, mean number of motile spermatozoa after sperm preparation, morphologically normal spermatozoa, and mean number of cleaved and transferred Oocytes and pregnancy rate were significantly higher in non-smoking patients group in comparison to smoking one (76.2±29.5mill/ml; 65.2±31.4%; 16.1±18.5%; 1.9±0.8%; 34%vs.50.7±39.2 mill/ml; 39.0±20.4%19.8±18.5%; 1.9±0.7% and 27.3% respectively). The other sperm parameters were similar in both groups. Cotinine concentration was significantly higher in smokers than non-smokers (144.3±82.2μg/dl vs.19.3±23.7μg/dl; p=0.001). Inverse correlations were found between Cotinine concentration and ejaculate volume, sperm concentration, motility, vitality, morphology membrane integrity, DNA fragmentation (TUNEL), TAS concentration and fertilization. Besides, a positive correlation was shown between Cotinine and ROS concentration. In the smoker group three patients become pregnant (27.3%) and 11 patients achieved pregnancy in the non-smoking group (34%; p=0.001). CONCLUSIONS: Cotinine concentration in seminal plasma has an adverse effect on seminal plasma anti oxidants, sperm parameters and ART outcome and increase the production of ROS in seminal plasma. Therefore, it is recommended to quit smoking in childless patients who are seeking ART therapy.