Background: The increasing prevalence of infertility cases is becoming a major public health problem in developing countries due to changes in diet and lifestyle. Objectives: Black Carrot Extract BCE (Daucuscarota) plays an important role as fertility agent due to its antioxidant and anti-proliferative properties. Therefore, the aim of the present study was to identify the effects of different concentrations of BCE on the outcome of in vitro human sperm activation of infertile patients. Methods: Thirty two semen samples were prepared which served as a control, using direct swim-up technique. Modified Earl's medium (MEM) was used only in group 2 (G2) and two concentrations of BCE (10µg/1ml and 20µg/1ml) were added to culture media MEM of groups 3 (G3) and 4 (G4) for in vitro sperm activation (ISA). Results: A highly significant (P<0.001) correlation was noticed between G4 with G2 and G3 in progressive motility when activation by adding 20µg/1ml of BCE to culture media as compared to G3 (which adding 10µg/1ml of BCE to culture media) and G2 (activation with culture media only). Moreover, swim-up activation technique with culture media and by both adding (10µg/1ml and 20µg/1ml of BCE respectively) to culture media enhancement in the (sperm motility, progressive sperm activity and sperm morphology). Conclusion: The high concentration of BCE is effective in the enhancement of progressive sperm activity during in vitro human sperm activation.
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.
Background: The semen cryopreservation has an important role in the male fertility preservation. Certainly, patients who are prone to become infertile due to surgical or medical treatments such as chemo- or radiotherapy for cancer treatment. In fact, semen cryostorage seems to be the only proven method that may offer these couples a chance of having children in the future. Objectives: This study was aimed to investigate the effects of cryopreservation protocols on human sperm parameters. Methods: Sixty semen samples with range of age (19-35) years were included in this study. Sperm parameters, were assessed pre- or post-cryopreservation. Cryopreservation protocols were done using SMART medium with either 15% dimethyl sulfoxide(DMSO) or 15% glycerol alone (as control group) or supplemented with either 0.25M or 0.5M of sucrose for treated groups. Crude data were statistically analyzed. Results: Generally, the results of the present study showed that the sperm parameters were significantly deteriorated post-thawing (P<0.05) as compared to precryopreservation. Both treated groups (G2 and G3) showed best results as compared to G1 (control groups). In contrast, treated group (G2) with 0.25 M of sucrose appeared better results than the as G3 group that treated with 0.5M of sucrose. Conclusion: Sperm parameters including concentration, motility, and morphology were deteriorated after cryopreservation. Using 0.25M sucrose reduced the impact of cryopreservation.
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.
This study was designed to assess the effect of different concentrations of L-carnitine and Co enzyme Q10 (COQ10) supplied to SMART Pro-medium on human sperm motility and sperm grade activity ( progressive, non-progressive and immotile) during in vitro sperm activation. Eighty seven samples of semen of infertile and fertile men were randomly collected from the subjects at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies / Al-Nahrain University. The mean age of the men was (32.034 ± 0.57) years with mean duration of infertility (4.644 ± 0.25) years. Seminal fluid analyses were done involving macroscopic and microscopic examinations according to WHO criteria. Swim-up technique was dependent for in vitro sperm activation , the washed samples were divided before using centrifugation swim-up technique into 3 groups: control group (G1) without L-carnitine and CoQ10. While, in G2, G3 two concentrations of L-carnitine and CoQ10 (20 µg , and 40 µg ) were used. Moreover, all groups within post-activation enriched with SMART-Pro media. The sperm motility and sperm grade activity were evaluated after the addition of L-carnitine and COQ10. The results showed a significant (P<0.05) differences were showed in sperm motility and sperm grade activity of post-activation when compared with the pre-activation. In G2 and G3, with dose of ( 20 µg and 40 µg) of L-carnitine and CoQ10, the results showed a significant (P<0.05) increase in the percentages of sperm motility, progressive sperm motility in spite of the increase in the percentages of sperm motility, progressive sperm motility in G3 compared to G2, there was non significant (P>0.05) differences between G2 and G3. The results for non-progressive sperm motility% and total progressive sperm motility millions/ mL. for 40 µg ( G3) groups were significantly (P< 0.05) decreased when compared to control group (G1) with non-significant (P>0.05) differences between 20µg (G2) group, with significant (P< 0.05) decreased in immotile sperm percentage in post-activation when compared to the pre-activation. From the results of the study, it can be concluded that the addition of 40 µg of L-carnitine and CoQ10 to washed sperms can improve sperm motility and sperm grade activity in vitro. Data were analyzed statistically using complete randomized design (CRD) (one way ANOVA).
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.
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.
Background: One of the major concerns among the male reproductive health is the inability to achieve parenthood even after regular unprotected intercourse. The efficiency of the sperm selection techniques is expressed as the concen-tration of spermatozoa with normal motility. Discontinuous density gradient centrifugation (DGC) method widely used over the last decade due to effec-tiveness and reproducibility in recovering most motile sperm, which separates sperm based on their motility, size, and density differential. Glutathione act as an amino acid donor during spermatogenesis. If the levels of glutathione are too low during spermatogenesis, the number of mature and morphologically normal spermatozoa generated will be decreased. Objectives:The aim of this study was to assess the beneficial role of TAD 600mg Glutathione combined with DGC technique for activation of human sperm of asthenozoospermic, oligozoospermic and normozoospermic subjects compared with the DGC technique alone. Patients, Materials and Methods:Sixty males were involved in this study, di-vided into three groups, (twenty asthenozoospermic, Twenty oligozoospermic and twenty normozoospermic subjects)during their attendance to the Infertility Clinic at High Institute for Infertility Diagnosis and Assisted Reproductive Tech-nologies; Al-Nahrain University. Semen samples were obtained, and seminal fluid analysis was assessed. Each semen sample was divided into three parts. The first part prepared as in vitro sperm characterization before activation, the second part using density gradient centrifugation technique, while the last part prepared using density gradient centrifugation combined with TAD 600mg Glu-tathione. Results: After in vitro sperm activation for asthenozoospermic, oligozoospermic and normozoospermic samples a significant increase was observed in the sperm function parameters including sperm concentration, motility and morphological-ly normal sperm percentage as compared to pre-activation using combined tech-niques (density gradient centrifugation and TAD 600mg Glutathione) as com-pared to DGC technique alone. Conclusions:TAD 600mg Glutathione combined to the DGC technique was found to give higher significant results on sperm function parameters (sperm concen-tration, motility and morphology) when using a low quality of semen samples such as decreased sperm motility as compared with DGC technique alone.