For conception and the development of healthy embryos, sperm DNA integrity is crucial. According to a growing body of studies, there is a strong correlation between sperm DNA damage and male infertility. Among the new medicines being developed in the medical field, the application of Platelet Rich Plasma (PRP) in human reproduction has yet to be examined. A total of 100 semen samples were used in the current experimental investigation. From November 2020 to June 2021, the research was conducted at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies. Masturbation was used to get an ejaculated semen sample. After semen analysis, the samples were separated into two equal parts, one without autologous PRP and the other with 2% autologous PRP, with the DNA fragmentation assessed using the Sperm Chromatin Dispersion Test. There was highly significant reduction in DNA fragmentation index (p < 0.001). The mean sperm DNA integrity was reduced after adding PRP (33.85±16.73 vs 38.55±16.64), Mean (± SE). PRP has been shown to improve human sperm DNA integrity.
Background: Semen cryopreservation is a useful tool for preserving fertility for men involved in ART program who have been diagnosed with cancer and will undergo chemotherapy, radiotherapy or testicular surgery. Objective: The objective of this study to evaluate the effect of honey bee supplemented to cryoprotectant medium on post-thaw sperm DNA integrity ,morphology and round cells. Subjects ,Materials and Methods: Thirty semen samples were collected from 30 infertile patients. After assessment of semen analysis, semen samples were divided into 3 aliquots (0.7ml for each sample) and mixed with 1 ml of cryopreservation solution (G1,control) alone, enriched with 5% honey bee(G2) with 10% honey bee (G3) for cryopreservation. Current study used the acridine orange test to investigate the effect of honey bee in two concentration (5%, 10%) on the integrity of sperm chromatin structure pre-and post cryopreservation. Cryopreservation was done at -196 Co in liquid nitrogen and thawing was performed after six months. Direct swim up technique was used for in vitro sperm preparation post-thawing, sperm parameters were assessed and data were statistically analyzed pre-and post-thawing. Results: The results show there was a significant(P<0.05) increase in the percentage of morphologically normal sperm for all groups post-thawing, particularly for G3 group that was significantly (P<0.05) increased as compared to G1 and G2 groups, In contrast, non-significant differences (P>0.05) were observed between G1 and G2 groups. Sperm DNA fragmentation percentage (%) was significantly(P<0.05) increased in G1 post thawing without any additives as compared to pre-cryopreservation , where as in G2 and G3 sperm DNA fragmentation were increased but not significantly as post-cryopreservation groups. Also the results show that there were no significant differences between G2 and G3 whereas G3 gives better result as compared to G1 with significant differences. Round cells counts for all groups of post cryopreservation were significantly(P>0.05) decreased as compared to pre-cryopreservation. Non significant (P>0.05) differences in the round cells counts among all the groups of post -thawing in spite of G3 group has the lowest mean. Conclusions: From these results we concluded that supplementation of 10% of honey bee to the freezing cryoprotectant medium gives best protection to the morphology and DNA integrity of human sperm.
Background: Varicocele is an abnormal dilation of the testicular veins in the pampiniform plexus and characterized by retrograde blood flow in the internal spermatic veins due to incompetent or absent valves. It is considered one of the main causes of male factor infertility. Sperm DNA integrity is essential for in vivo and in vitro fertilization, and varicocele causes a progressive decrease in sperm quality . Objective: To study the impact of varicocele on semen quality and sperm DNA integrity. Patients, Materials and Methods: Twenty patients with varicocele and 20 men without varicocele were assigned into two groups in the Out-Patient Clinic of Urology in Al-Kadhimya Teaching Hospital . Semen was collected from both groups and the rate of sperm DNA fragmentation assessed by the "Comet assay" and categorized to 4 classes. Class I (no DNA fragmentation), class II (little DNA fragmentation), class III (meaningful DNA fragmentation), and class IV (high DNA fragmentation) . Results: No statistically significant differences were found between the varicocele and control groups with respect to age. Men with varicocele had significantly higher ejaculate volume, lower sperm concentration, and lower progressive motility, and showed more abnormal morphology than men in the control group. The Varicocele group showed a lower percentage of sperm with little DNA fragmentation (class II) and a higher percentage of sperm with DNA fragmentation (class IV), than the control group. Conclusions: Compared with men without varicocele, men with varicocele had a higher percentage of cells with DNA fragmentation .
Background: Many studies conducted on the pentoxifylline (PX) and Glycyrrhiza glabra (G.glabra) as motility stimulants showed positive effect on sperm preparation in vitro improving the progressive forward movement. However, it was not known the impact of these stimulants on the nature of genetic material, especially after cryopreservation. Objective: The experiment was designed to find out any harmful effect of a medium containing a mixture of PX and aqueous extract of G.glabra on the sperm DNA before and after cryopreservation. Materials and Methods: Fifty mature fertile male mice 8-12 weeks old were used. Epididymal sperms were obtained from caudal region and prepared in vitro by direct activation technique using four media namely; PX alone, G.glabra alone, a mixture of PX with G.glabra and Ham's F-12 (as control medium). Certain sperm function characteristics were examined as well as evaluation of DNA integrity by acridine orange test and comet assay before and after cryopreservation. Results: The results showed a highly significant (P<0.001) improvement in the sperm forward movement, with a highly significant (P<0.001) decrease in DNA abnormality using acridine orange test and comet assay, following in vitro activation by the four media compared to before activation. Following in vitro activation and cryopreservation, the results of epididymal sperm showed a highly significant (P<0.001) decrease in DNA abnormality by using PX medium alone, G.glabra medium alone and the mixture of PX and G.glabra compared to control medium and the results before cryopreservation. Conclusion: According to the results of present study, it has been found no toxic effect of medium containing PX and/or G.glabra on the epididymal sperms DNA after activation in vitro and cryopreservation using acridine orange test and comet assay. The best result obtained when using a medium containing a mixture of PX+ G.glabra together. These data could be applied for preparation and activation of epididymal and testicular sperms obtained from obstructive azoospermic men.
Human sperm chromatin and integrity of DNA have multiple endogenous factors, such as sperm compression and nucleoproteins, with various mechanisms all through their own sperm production and transport. The integrity of the paternal genetic material in both regular and assisted pregnancy is essential for initiating and maintaining a viable pregnancy. The study included semen samples from 100 men taken from Al Nahrain University Medical Clinic and Babylon Teaching Hospital for Women and Children. The first sperm test was based on the World Health Organization (1999), followed by a DNA integrity test that uses a DNA fragmentation test, and the third intensification test Chromatin by aniline blue dye. It was correlated negatively with the sperm count in combination with DFI and chromatin sperm with semen characteristics (r = - 0.223, p = 0.026). As well as DFI, sperm chromatin was being clinically significant associated with the rate of round cells (r = 0.39, < 0.001). DFI was positive and significantly associated with sperm chromatin in all infertile men (r = 0.958; p<0.001). DNA isolation in all enrolled men has been adversely linked with sperm/ejaculation, has been associated with the round cell count positive way and is not understanding and resolving with other parameters. The width of abstinence and beneficial cell count in all register men were strongly linked with Sperm chromatin. There was a direct relationship between the fragmentation of DNA and chromatin. This could imply that one exam can be used instead of the other.
When evaluating male reproductive potential, it is crucial to evaluate sperm structural abnormalities and sperm DNA damage. Apoptosis and mitochondrial membrane potential disruption are linked to sperm DNA fragmentation (SDF), which is inversely correlated with motility and normal morphology. To enable liquefaction, the semen sample is taken in a sterile collection container and incubated at 37°C for 30 minutes The samples were then split into three groups, P, G, and N, based on the morphological %. To measure chromatin condensation, aniline blue staining (AB), an acidic dye with a strong affinity for non-lysine-rich histones, was employed. The DNA maturity of the spermatozoa was assessed using AB. Sperm concentration, total sperm count, total motility, and total progressively motile sperm all had means that were above the WHO 2010 standard range. The DNA fragmentation index was significantly higher in patients with pattern P than in patients with patterns G and N, and it differed significantly (P=0.003) between the semen morphological patterns. In conclusion, as DNA fragmentation is an independent test this study showed that a low percentage of normal sperm morphology might indicate DNA integrity status.
The ability of human sperm to fertilize can be compromised both in vivo and in vitro by the toxins found in cigarette smoke, which can lower sperm mitochondrial activity and harm chromatin structure. The aim is to study the effect of smoking on the apoptotic marker (caspase-3) of human sperm. A cross-sectional study of 80 semen samples of sub-fertile men, were divided into smokers (35 patients) and non-smokers (45 patients). All the semen samples after seminal fluid analysis underwent HOST test and grading (A-G) according to the WHO 2021 criteria then the slides were smeared and stained with an immunocytochemistry kit for Caspase-3 apoptotic marker. There was no significant effect of smoking in the concentration of caspase-3 expression, in spite of the high presentation of both grades A and G in the smoker group ( p = 0.57, 0.56 respectively). Smoking has a negative impact on the DNA integrity of sperm as it increases the apoptotic marker within the smoker group.
Exposure to radiation of various forms is one of the many environmental, health, and lifestyle variables that have been linked in recent years to an increase in the rate of male infertility. According to data from several recently published studies, mobile phone use may negatively affect sperm parameters, which may lower male fertility. Ninety-seven samples of semen from fertile and infertile individuals were sent to the "High Institute for Infertility Diagnosis and Assisted Reproductive Technologies. Al-Nahrain University in Baghdad, between November 2022 and May 2023. The samples were split into four groups: normozoospermia (n = 40), asthenozoospermia (n = 25), oligozoospermia (n = 16), and teratozoospermia (n = 16). Each group was further divided into two groups based on the amount of time spent using a mobile phone each day: less than 4 hours per day and equal to or more than 4 hours per day. A routine seminal fluid analysis was carried out to measure the sperm parameters and test the deoxyribonucleic acid fragmentation rate using an acridine orange stain. Samples exposed to electromagnetic waves of cell phones showed a significant decrease in DNA normality. deoxyribonucleic acid fragmentation index (p < 0.001) in normozoospermia patients exposed to more than 4 hours/day of non-ionizing electromagnetic waves. The results of deoxyribonucleic acid fragmentation index after treatment were shown a significant improvement compared to before the treatment in all fertile and infertile groups. Electromagnetic waves emitted from cell phones are associated with a decrease of deoxyribonucleic acid normality when exposure for <4hours/day. The study recommend that men of reproductive age avoid keeping their cell phones in the hip pocket and front pants pocket which may negatively affect spermatozoa deoxyribonucleic acid integrity and impair male fertility.
One-half of the genome is delivered by the sperm, and a healthy sperm is needed for fertilization, fetal development, and child wellbeing. Abnormal sperm DNA can impact any of these processes. The measurement of sperm DNA fragmentation has been added to some medical andrology laboratories as a crucial addition to the standard analysis of the semen. Sperm DNA fragmentation has also been linked to male infertility including idiopathic male infertility. Some degree of controlled DNA nicking is essential for adequate DNA compaction, but excessive sperm DNA fragmentation is usually associated with reduced male fertility potential, reduced fertilization, poor embryo quality, recurrent pregnancy loss and poor assisted reproductive techniques outcomes. Semen samples from 50 infertile males (as 19 asthenozoospermia, 7 oligozoospermia, 13 normospermia and 11 combined defects) were analyzed for sperm DNA fragmentation. This study followed the World Health Organization 2010 guidelines to assess the sperm parameters. Sample were collected by masturbation and put in a 37 °C incubator to be liquefied, then they were placed in a centrifuge (3000 rpm for 10 min) to extract the pellet, which was used for assessment of DFI% by anillin blue staining. Negative significant correlation (p = 0.023) between morphologically normal sperms and DFI % was found in normozoospermia group. The objective of this study is to examine the correlation of DFI among different seminal fluid categories (normozoospermia, oligozoospermia, asthenozoospermia and, combined).
The integrity of sperm deoxyribonucleic acid (DNA) and chromatin is very important for genetic material transmission into offspring. Therefore, the aim of the present study was to investigate the correlation between sperm deoxyribonucleic acid integrity and sperm parameters. The current study was conducted on 96 semen samples. This study was done at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University, for the period between November 2020 and May 2021. The samples were collected, and seminal fluid analysis was performed according to World Health Organization guidelines. Sperm Chromatin Dispersion test was performed. The results showed that the deoxyribonucleic acid fragmentation of sperms in infertile men in this study was higher than reported in fertile men in previous studies, indicating deoxyribonucleic acid fragmentation (DNA) is actively contributing at least partially to male infertility. In terms of enrolled men, sperm deoxyribonucleic acid fragmentation (SDF) showed a significant positive correlation with round cell count.