The role of sperm DNA integrity in couples with recurrent implantation failure following IVF treatment.

There is increasing evidence from the literature that indicates the association between impaired sperm DNA integrity and male infertility. However, the data is insufficient regarding recurrent implantation failure (RIF) and sperm DNA damage. This study aimed to investigate the association between sperm DNA fragmentation and RIF cases. Basic semen parameters and sperm DNA fragmentation index (DFI) of men whose partner was suffering from RIF were compared with men whose partner was diagnosed with unexplained infertility (UEI) but had clinical pregnancies following IVF treatment. A retrospective analysis from a large-volume IVF center has been performed, and a total of 197 couples underwent analysis. Two groups were formed, couples with RIF and couples diagnosed with UEI but had clinical pregnancies (controls) following IVF cycles. The mean number of cycles showed significant differences between the groups. However, no statistical difference was observed between RIF and the control group regarding patient characteristics, semen parameters, and sperm DNA fragmentation index (DFI). Also, no statistically significant correlation was found between sperm DFI and clinical pregnancies in the unexplained infertility cohort. Our results show that sperm DNA fragmentation may not be an important contributing factor to RIF cases.

Recovery of fertility in azoospermia rats after injection of adipose-tissue-derived mesenchymal stem cells: the sperm generation.

The recent reports on the treatment of azoospermia patients, in which spermatozoa could not be traced in their testes, are focused more on the potential use of adult stem cells, like mesenchymal stem cells (MSCs). The aim of this study was to demonstrate the potential use of MSCs derived from adipose tissue in the treatment of azoospermia using rat disease models. After busulfan application, the rats (n = 20) were injected with the GFP(+) MSCs into left rete testes. After 12 weeks, the testes with cell injection (right testes) were compared to control (left testes) after dimensional and immunohistochemical analyses. Testes treated with MSCs appeared morphologically normal, but they were atrophic in rats without stem cell treatment, in which the seminiferous tubules were empty. Spermatogenesis was detected, not in every but in some tubules of cell-treated testes. GFP(+)/VASA(+) and GFP(+)/SCP1(+) cells in testes indicated the transdifferentiation of MSCs into spermatogenetic cells in the appropriate microenvironment. Rats with cell treatment were mated to show the full recovery of spermatogenesis, and continuous generations were obtained. The expression of GFP was detected in the mesenchymal stem cells derived from adipose tissue and bone marrow and also in the sperms of offspring. In conclusion, MSCs might be studied for the same purpose in humans in future.

A new efficient method for eliminating the interference effect of human serum and increasing the sensitivity and recovery rate of enzyme immunoassay.

A new, very simple method for increasing the sensitivity and recovery rate of enzyme-linked immunosorbent assay (ELISA) for the precise quantification of antigen in human serum is described. The assay design uses CATNF6A4c IgG2a monoclonal antibody and biotinylated anti-human tumor necrosis factor-alpha (hTNF-alpha) polyclonal mouse IgG as the capture and tracer antibodies, respectively. The assay is completed within 4 hours at room temperature and is capable of detecting both recombinant and native human TNF-alpha. The assay incorporates the use of saturated ammonium sulfate (SAS) as a component of the dilution buffer to amplify the resultant signal from antigen containing human serum and eliminating the endogenous interference of native human serum. SAS worked optimally at the final concentrations, ranging from 1.2% to 11%. The addition of SAS to the dilution buffer resulted in a dramatic increase in both sensitivity and recovery rate of the ELISA. The results demonstrated that 50 microL of dilution buffer, containing SAS, enabled the precise quantification of human TNF-alpha in 100 microL of human serum samples and eliminated the interference of native serum, which seemed to be related to complement proteins. Therefore, dilution buffer containing SAS, at a defined concentration, seemed to be a potential candidate for resolving sensitivity and recovery problems usually encountered in immunoassays when measurement was performed with native serum samples. The proposed technique provides an easy, practical, and consistent method for ELISA when using human native serum.