Prolonged semen incubation alters the biological characteristics of human spermatozoa.

OBJECTIVE: The present study assessed the biological characteristics of human spermatozoa at different time intervals (0, 1, 1.5, and 2 hours) after incubation at 37°C. METHODS: Twenty-five normozoospermic semen samples were incubated at 37°C. Incubation was performed at four time intervals of 0 (after liquefaction), 1, 1.5, and 2 hours. The samples were evaluated for sperm parameters at each time interval. RESULTS: The rate of sperm progressive motility decreased at 1.5 hours compared to 0 hours as well as 2 hours compared to 1 hour and 0 hours. The rate of non-motile spermatozoa also decreased after 2 hours compared to after 0 hours. No significant changes were observed in sperm viability (p=0.98) and non- progressive motility (p=0.48) at any time intervals. Abnormal sperm morphology increased at 1.5 hours of incubation time (p<0.001). No significant changes were observed in DNA fragmentation at 1 hour compared to 0 hours (median [interquartile range]: 19.5 [4] vs. 19 [4]), as well as at 1.5 hours compared to 1 hour (20 [5]). However, a significant increase in DNA fragmentation was observed at 1.5 hours compared to 0 hours. The mitochondrial membrane potential decreased remarkably after 1 hour of incubation time. No significant differences were observed in the acrosome reaction or malonaldehyde levels at any time point (p=0.34 and p=0.98, respectively). CONCLUSION: The incubation of normozoospermic samples before use in assisted reproductive technology should be less than 1.5 hours to minimize the destructive effects of prolonged incubation time on general and specific sperm parameters.

Prolonged exposure of human spermatozoa in polyvinylpyrrolidone has detrimental effects on sperm biological characteristics.

Polyvinylpyrrolidone (PVP) has been utilized in intracytoplasmic sperm injection (ICSI) for immobilization and manipulation of spermatozoa. This study aims to determine the suitable time that sperm cells could be safely exposed to PVP during ICSI procedure. Twenty-five normal semen samples were prepared using the swim-up method and then were exposed to 10% PVP at different time intervals (15, 30 and 60 min). The effect of PVP on sperm parameters (viability and morphology), DNA fragmentation index (sperm chromatin dispersion test), chromatin quality (aniline blue, toluidine blue and chromomycin A3 staining), acrosome reaction, mitochondrial membrane potential and sperm ultrastructure was assessed at different time intervals. Our results showed that prolonged sperm exposure in PVP for 15, 30 and 60 min significantly affects viability and morphology with a concomitant increase in DNA fragmentation and abnormal chromatin structure, while the percentage of acrosome-reacted spermatozoa was additionally increased. In addition, the spermatozoa with high mitochondrial membrane potential were significantly decreased compared to unexposed spermatozoa to PVP. In conclusion, the detrimental effects of PVP were increased significantly following sperm exposure in PVP after 15 min. Therefore, the sperm exposure to PVP should be limited to less than 15 min during ICSI procedure.

Microfluidic sperm selection yields higher sperm quality compared to conventional method in ICSI program: A pilot study.

An appropriate preparation technique, should be capable of isolating highquality spermatozoa for intracytoplasmic sperm injection (ICSI). The aim was to assess sperm quality parameters, DNA integrity, embryo development, and clinical outcomes using a practical and accessible Microfluidic Sperm Sorting (MSS) technique. A total of 95 ICSI cases performed using sperm samples were prepared with our MSS (group 1) or by Direct Swim Up (DSU; control) method (group 2). Both sperm quality parameters and sperm DNA fragmentation (SDF) were compared between the groups. DNA fragmentation was assessed using Sperm Chromatin Dispersion (SCD) test and fine morphology was assessed using Motile Sperm Organelle Morphology Examination (MSOME). Embryo development and clinical outcomes were compared between the groups. In the MSS group, progressive motility and the fraction of Class I sperm morphology sperm were significantly higher compared to DSU group (P < 0.01 and P < 0.001, respectively). Moreover, the rates of DNA fragmentation and immotile spermatozoa were significantly lower in MSS when compared to DSU group (P < 0.001). Also, higher rates of high-quality embryo formation (P < 0.001), implantation (P = 0.04) and pregnancy (P = 0.05) were achieved in the MSS compared to DSU groups. The MSS technique proved to be a noninvasive, disposable, easy to use, and inexpensive method for separation of high-quality spermatozoa. Both laboratory parameters and clinical outcomes were improved with application of MSS for neat sperm collection in ICSI.AbbreviationsICSI: Intracytoplasmic Sperm Injection; MSS: Microfluidic Sperm Sorting; Sperm DNA Fragmentation (SDF); SCD: Sperm Chromatin Dispersion; MSOME: Motile Sperm Organelle Morphology Examination; DGC: Density Gradient Centrifugation; DSU: Direct Swim Up; ROS: Reactive Oxygen Species; ART: Assisted Reproducetive Technology.

The Determination of Estradiol to Cumulus Oocyte Complex (COC) Number Ratio: Does it Predict the Outcomes of ART Cycles?

BACKGROUND: The aim of this study was to assess the impact of total serum E2 on the day of human chronic gonadotropin (hCG) administration and the serum E2 per oocyte ratio on the outcomes of assisted reproductive technology (ART) cycles. METHODS: A total of 205 women were categorized into 3 groups according to the serum E2 levels: 1: ≤1500 pg/ml; 2: 1500-3000 pg/ml; 3: >3000 pg/ml. Another categorization included 3 groups according to E2/oocyte ratio: A: ≤150 pg/ml per oocyte; B: 150-200 pg/ml per oocyte; and C: >200 pg/ml per oocyte. The outcome compared between groups included laboratory and clinical characteristics. One-way analysis of variance (ANOVA), chi-square and Kruskal-Wallis, and multiple logistic regression model were performed, and appropriate differences were considered significant at p<0.05. RESULTS: There was a significant difference between the groups based on the E2 levels with respect to laboratory parameters. In group C, the rates of chemical pregnancy (54.1%), clinical pregnancy (50%) and live birth (45.8%) were significantly higher, when compared to other groups. Moreover, according to E2/oocyte ratio, the rate of live birth was higher in group C compared with group A (18.3%, p=0.04), and group C (29.7%, p<0.0001). Logistic regression showed the number of good quality embryos was a positive predictor for live birth (odds ratio=2.03, 95% CI=1-4.1), but the level of E2 on day of HCG was a negative predictor (odds ratio=0.99, 95% CI=0.99-1). CONCLUSION: Supraphysiological levels of E2 had no adverse effects on the quality of the embryos in IVF cycles, but may have adverse effect on live birth in fresh transfer. Also, it is confirmed that both the pregnancy and live birth rates were elevated with E2/oocyte ratio ≥200 pg/ml.

Administration of high dose of methamphetamine has detrimental effects on sperm parameters and DNA integrity in mice.

BACKGROUND: Methamphetamine (MA) was shown to have harmful effects on male reproductive system. OBJECTIVE: To investigate probable effects of daily administration of MA on sperm parameters and chromatin/DNA integrity in mouse. MATERIAL AND METHODS: Thirty-five NMRI male mice were divided into five groups including low, medium, and high dosage groups which were injected intraperitoneally with 4, 8 and 15 mg/kg/day for 35 days, respectively. Normal saline was injected in sham group and no medications were used in control group. Then, the mice were killed and caudal epididymis of each animal was cut and placed in Ham's F10 medium for sperm retrieval. To evaluate sperm chromatin abnormalities, the aniline blue, toluidine blue and chromomycine A3 were used. For sperm DNA integrity and apoptosis, the acridine orange, sperm chromatin dispersion, and TUNEL assay were applied. For sperm morphology, Papanicolaou staining was done. RESULTS: Normal morphology and progressive motility of spermatozoa decreased in medium and high dosage groups in comparison with the control group (p=0.035). There was a significant increase in rate of aniline blue, toluidine blue, and chromomycine A3 positive spermatozoa in high dosage group. In a similar manner, there was an increase in rates of acridine orange, TUNEL and sperm chromatin dispersion positive sperm cells in high dosage group with respect to others. CONCLUSION: MA abuse in a dose-dependent manner could have detrimental effects on male reproductive indices including sperm parameters and sperm chromatin/DNA integrity in mice.