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Objective: Sperm DNA fragmentation and maturation directly interferes with reproductive efficiency. Although there are several methods for assessing sperm DNA integrity, however, many of them are laborious and require high-precision equipment in the clinics. Thus, evaluating economic and reliable methods to prepare suitable sperm for assisted reproductive technologies without DNA damage is critical., Material and Methods: A total of 114 semen samples were collected and analyzed using computer-assisted semen analysis. The DNA fragmentation index (DFI) of all samples was evaluated by two methods of sperm chromatin dispersion (SCD) and sperm chromatin structure assay (SCSA). Besides, chromatin maturation index (CMI) was assessed by three methods including aniline blue (AB)-sperm chromatin maturation assay (SCMA), fluorescence microscopic chromomycin A3 (fmCMA3), and flow cytometric CMA3 (fcCMA3)., Results: The result showed that the DFI had no statistically significant differences (p>0.05) between SCSA (26.98%±1.28%) and SCD (27.88%±1.278%), although SCD demonstrated a strong correlation with DNA maturity (p<0.001), which had not been seen in SCSA. Besides, the CMI demonstrated significant differences (p<0.001) when assessed by AB-SCMA (14.86%±0.65%), fmCMA3 (29.18%±1.01%), and fcCMA3 (22.45%±0.62%). Among these, only the fmCMA3 showed a significant correlation with semen parameters (p<0.01) and embryo development (p<0.001)., Conclusion: It seems that SCD and fmCMA3 were more accessible, affordable, and reliable tests for assessing DFI and CMI. It appeared these two methods may be the best choices for evaluating sperm DNA integrity in clinics.

Objective: Studies showed a decrease of the semen analysis parameters and an increase in the average age of first-time fathers over the past several decades. The aim of the present study was to assess the influence of paternal age on semen quality and fertilization outcomes in men with normal sperm DNA fragmentation and chromatin maturation index (DFI and CMI), reactive oxygen species (ROS), and total antioxidant capacity (TAC) levels., Material and Methods: The study was performed on 70 men with their wife's age ≤38 years and normal sperm DFI, CMI, ROS, and TAC levels. None of the couples had a history of genital inflammation, chronic diseases, endocrine abnormality, chromosomal aberrations, Y chromosome microdeletion, azoospermia, and leukocytospermia. These men were separated into 2 groups according to their age (group A: age <45 years and group B: age ≥45 years). Semen analysis and fertilization outcome after using the intracytoplasmic sperm injection were assessed in both groups., Result: Sperm concentration showed a significant reduction in group B (p=0.04). Although semen volume, sperm normal morphology, and progressive motility were decreased in group B, the reduction was not significant when compared with group A (p=0.09, p=0.47, and p=0.77, respectively). In addition, the differences of embryo quality with grades A, B, and C and 8-cell embryo formation were not statistically significant between the 2 groups., Conclusion: These results demonstrated that in men with normal sperm DFI, CMI, ROS, and TAC levels, there were no significant changes in semen parameters and fertilization outcomes with an increasing age.

Objective: Fibronectin (FN) is a multifunctional diametric glycoprotein on the surface of the sperm that plays an important role in the sperm-oocyte interaction and fertilization process. The aim of the present study was to assess the FN levels as a sperm surface biomarker for sperm selection in assisted reproductive technology., Material and Methods: Polyclonal antibody against human FN was produced in rabbit. Its quality, purity, and immune reactivity were assessed by SDS-PAGE and western blot. In addition, the presence of FN on the sperm surface was assessed through immunocytochemistry and flow cytometry. The amount of FN and the sperm quality were assessed in normozoospermia (N) (42 men) and asthenoteratozoospermia (AT) (72 men) groups through sperm chromatin dispersion (SCD), sperm chromatin structure assay (SCSA), and chromatin maturation index (CMI)., Results: The results showed the distribution of FN protein on the equatorial region of the human sperm surface. In addition, the FN levels were found to have a significant difference between the two groups with 24.64±9.08% in N and 16.90±7.27% in AT (p≤0.0001). In addition, FN level negatively correlated with SCD (p≤0.0001), SCSA (p≤0.0001), and CMI (p≤0.001). Threshold values of FN level and DNA fragmentation index (DFI) percentage were 16 and 30 and were identified as cut-off values to determine the N group with a specificity of 83.3% and 81.0% and a sensitivity of 16.8% and 19.0%, respectively. The specificity and sensitivity of FN-DFI were 91.2% and 8.8%, respectively., Conclusion: It appears that FN can be used for the selection of sperm with suitable quality, although future studies are recommended.