Coadministration of calcium chloride with lead acetate can improve motility of cauda epididymal spermatozoa in Swiss white mice

Background: Lead is an industrial heavy metal that can decrease sperm motility. Objective: The aim was to investigate the protective effects of calcium against lead on motility of spermatozoa. Materials and Methods: In total 40 adult male Swiss white mice were randomly divided into 5 groups (control, lead of 1st wk, lead of 2nd wk, lead/calcium of 1stwk and lead/calcium of 2nd wk). The lead groups of mice were injected by a single dose of lead acetate (200 mg/kg) intraperitoneally. Lead/calcium groups of mice were injected by a single same dose of lead acetate along with three doses of 80 mg/kg calcium chloride. The control group of mice was injected only with same volume of distilled water through the same route. Mice of 1st and 2nd wk groups were sacrificed through cervical dislocation one and two weeks after injections respectively. Results: Mean of the progressive motile spermatozoa of cauda epididymis in lead/calcium group of the first week was higher than the lead group of the first week and this difference was significant. There was not any significant difference among weight of testes and epididymides of all groups. Conclusion: It can be concluded that calcium can decrease the effects of lead on sperm motility. © 2016, Research and Clinical Center for Infertitlity. All rights reserved

Effects of temperature and storage time on the motility, viability, DNA integrity and apoptosis of processed human spermatozoa

The aim of this study was to evaluate motility, viability, DNA integrity and apoptosis of spermatozoa when washed semen samples were kept for up to 12 days at 4–6°C and 25°C. In this experimental study, 26 normozoospermic semen samples were washed twice in Modified Ham's F10 and resuspended in IVF fertilisation medium. Half of the specimens were stored at 4–6°C, and the other half was kept at 25°C for 12 days. The proportions of viable, motile, spermatozoa with double-stranded DNA and apoptotic spermatozoa were examined during storage time. Apoptosis was measured using annexin V-PI staining followed by flow cytometry. Results showed that sperm motility and viability decreased during 12 days of sample storage (p < .001). There was no significant difference between the two temperatures in terms of motility and viability for up to 2 days (p < .05)

The effects of protamine deficiency on ultrastructure of human sperm nucleus

Background: Chromomycin A3 (CMA3) staining is one of the staining methods for detecting protamine deficiency in sperm nucleus. CMA3 is a fluorochrome that competes with protamines for binding to DNA double helix. It has been shown in our previous studies that percentage of CMA3 positive spermatozoa in semen has a close significant relationship with the fertilization rate in in vitro fertilization (IVF). The aim of this study was to examine the ultrastructural differences between sperms in patients who had high fluorescent percentages of yellow or red in CMA3 staining (protamine deficient) with patients with low fluorescent percentages. Materials and Methods: Semen samples are taken from five patients with high fluorescent percentages and five patients with low fluorescent percentages. Then the samples are passed for the different steps of preparing for electron microscopy. After the sectioning and mounting on grids, they are investigated under the transmission electron microscope. Results: Sperms in patients with low percentages of positive spermatozoa often have a normal appearance. Sperms in high fluorescent samples frequently have unpacked chromatin. Furthermore acrosomes of these sperms are thinner or disturbed. Also sometimes there are irregularities in sperm head membrane. Conclusion: Protamine deficiency in sperm nucleus can cause ultrastructural anomalies in sperm chromatin such as unpacking of it. It also is concomitant with acrosome and sperm membrane disturbances

Impact of sperm chromatin evaluation on fertilization rate in intracytoplasmic sperm injection

Background: Sperm DNA in human beings and most vertebrates is packed by protamines into highly compact form of chromatin. There are many staining methods to assess sperm chromatin. Three different methods of staining were used simultaneously in this study and the goal was to determine which of these sperm tests has a relation with fertilization rate in intracytoplasmic sperm injection (ICSI). Materials and Methods: Thirty couples who referred to Yamagata University Hospital (Yamagata, Japan) for ICSI were included in this study. The greater part of semen was prepared for ICSI. The remaining part was used for staining with aniline blue, acridine orange, and chromomycin A3 (CMA3). For evaluation of abnormal morphology and abnormality of head, Papanicolaou-stained smears were used. The analysis of data was done using Spearman coefficient of correlation and logistic regression model. Receiver operator characteristic (ROC) curve was used for discrimination of CMA3 staining power to identify ICSI rates. Results: Percentage of CMA3 positivity, unlike those of aniline blue and acridine orange, showed significant negative correlation with fertilization rate. Moreover, the percentage of CMA3 positivity showed a positive correlation with the percentage of abnormal morphology and abnormality of head. By dividing patients into CMA3 48% groups, the area under the curve was 0.646. Conclusions: CMA3 staining (protamine deficiency) could be considered as a useful tool for evaluation of male fertility prior to infertility treatment

The epididymal sperm viability, motility and DNA integrity in dead mice maintained at 4-6oC

Background: When male animals die, spermatozoa within the body of animal will be degenerated. Because of unique chromatin structure of sperm, maybe this degeneration is different from other cells. However there is not any research which considered directly the integrity of sperm DNA by keeping the cadaver in refrigerator. Objective: The aim of this study was to assess viability, total motility and DNA integrity of sperm cells after death. Materials and Methods: In this experimental study, 24 male Swiss white mice were killed by cervical dislocation and then kept in refrigerator (4-6oC) for up to 12 days. On the 0 (immediately after death as control group), 1st, 2nd, 3rd, 5th, 7th, 10th and the 12th days after death cauda epididymides were removed and squeezed in Ham’s F10 medium. The proportion of viable, motile and double stranded DNA spermatozoa was examined. Viability and DNA integrity of sperm cells were examined consecutively by eosin nigrosin and acridine orange stainings. Results: The data obtained from this study showed that viability and total motility of sperm cells were significantly decreased during 12 days after death (p<0.001). In contrast with viability and motility, DNA integrity was without significant changes (even 12 days after death). Conclusion: This study suggests that integrity of sperm DNA would not change even after 12 days after death if the cadaver kept in refrigerato

Thymoquinone as a natural spermostatic substance in reproductive medicine: An experimental study

Background: Nonoxynol-9 a nonionic surfactant is widely used for its spermicidal effects. Finding new sperm immobilizing agents is necessary because Nonoxynol-9 damages the tissues of female reproductive system. Objective: The aim of this study was to evaluate the effects of Thymoquinone (TQ) as a potential spermostatic compound on the motility and viability of human spermatozoa. Materials and Methods: In this experimental study, the effects of 5, 10, 20, 50, 100 &mu;g/ml, 1 and 10 mg/ml of TQ on normozoospermic semen samples were investigated. Sperm motility and viability were compared between untreated and TQ-treated aliquots of each semen sample. To evaluate the effects of TQ on the alteration of mitochondrial membrane potential (MMP), 32 semen samples were examined using 50 &mu;g/ml of TQ. Flow cytometric analysis was performed after staining of spermatozoa with JC-1. Results: Doses above 20 &mu;g/ml of TQ could eventually immobilize all spermatozoa in culture medium. Adding 50 &mu;g/ml of TQ did not significantly diminish the percentage of viable spermatozoa and flow cytometry results revealed that this amount of TQ could decrease sperm MMP. Conclusion: TQ could discontinue the movement of sperm cells in medium without reducing the population of live spermatozoa. It is more likely that TQ exerts its spermostatic action by mitigating the MMP of spermatozoa. Therefore, TQ could be considered as a potential new natural spermostatic chemica

Comparison of the efficacy of Piascledine and transforming growth factor β1 on chondrogenic differentiation of human adipose-derived stem cells in fibrin and fibrin-alginate scaffolds

Objective(s):The aim of this study was to compare the chondrogenic induction potential of Piascledine and TGF-β1 on adipose-derived stem cells (ADSCs) in fibrin and fibrin-alginate scaffolds.  Materials and Methods: Human subcutaneous adipose tissues were harvested from three patients who were scheduled to undergo liposuction. Isolated ADSCs were proliferated in a culture medium. Then, the cells were seeded in fibrin or fibrin-alginate scaffolds and cultured for 14 days in a chondrogenic medium containing Piascledine, TGF-β1, or both. The rate of cell proliferation and survival was evaluated by using MTT [3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide] assay and the rate of the expression of type II collagen, aggrecan, and type X collagen genes was evaluated by real-time polymerase chain reaction (real-time PCR) method. Results: The MTT results showed that Piascledine is able to enhance the proliferation and survival of ADSCs in fibrin scaffolds in comparison to other groups (