The functionality of human spermatozoa is a key factor for the success rate of natural human reproduction, but unfortunately the infertility progressively increases due to multifarious environmental factors. Such disquieting statistics requires the employment of sophisticated computer-assisted methods for semen quality analysis, whose precision, however, is unreliable in cases of patients with low sperm concentrations. In this study, we report a novel quartz crystal microbalance (QCM) based biosensor for in-situ quality assessment of male gametes, comprising a superhydrophobic soot coating as an interface sensing material. The soot deposition on the surface of a 5 MHz QCM eliminates the noise that normally arises upon immersion of the uncoated sensor in the test liquid environment, allowing the detection of human spermatozoa down to 1000–100,000 units/mL (1–100 ppb). Furthermore, the soot coated QCM delimitates in a highly repeatable way the immotile and motile sperm cells by inducing fundamentally distinct responses in respect to sensor sensitivity and signal trends. The obtained results reveal the strong potential of the superhydrophobic QCM for future inclusion in diverse laboratory analyses closely related to the in vitro fertilization procedures, with a final aim of gaining practical approaches for diagnoses and selection of male gametes.
OBJECTIVE: To investigate the effectiveness of immobilized acid-solubilized zonae pellucidae in the selection of spermatozoa for intracytoplasmic sperm injection (ICSI).DESIGN: A prospective sibling oocytes study. MATERIALS AND METHODS: In this study were included 113 couples who fulfilled the inclusion criteria: 1) unexplained infertility factor; 2) good quality oocytes; 3) fertilization failure for 3-5 consecutive ICSI procedures; 4) at least one oocyte at germinal vesicle stage (GV) and 5) at least four metaphase II oocytes retrieved during follicular puncture. Zonae pellucidae were isolated from the patient's own GVs. Zonae were acid solubilized and diluted in carbonate buffer (pH 9.6) for air dry immobilization on glass petri dishes. The partner's semen was washed and placed in the dishes. The spermatozoa that adhered on the immobilized surface were used for ICSI in the half of the retrieved oocytes from each woman. The other half of the oocytes was fertilized by conventional ICSI. In total, 312 oocytes were injected with zonaselected spermatozoa (zona-selection group) and 366 oocytes were injected with conventionally-selected spermatozoa (control group). The resulted embryos from the zona-selection and the control group were used in 43 and 50 single embryo transfers, respectively. Main outcomes were fertilization rate, embryo quality, implantation rate and pregnancy rate. Statistical analysis was performed using SPSS Software ver.21.RESULTS: Slightly higher fertilization rate was observed among the oocytes injected with zona-bound spermatozoa in comparison to the conventional ICSI group (75.6% vs. 72.3%, p¼0.38). Also no significant differences were observed in the embryo quality and in the implantation rates between the zona-selection and the control group (p¼0.24 and p¼0.59, respectively). However, the pregnancy rate was considerably higher in the zona-selection group when compared with the control group (34.8% vs. 16.4%, p¼0.02). Moreover the miscarriage rate also differed significantly (7% in zona-selection vs. 18% in control group, p¼0.03).CONCLUSIONS: The use of patient's zona pellucida immobilized proteins in selection of spermatozoa for ICSI increases pregnancy rates and reduces the risk of miscarriage in couples with unexplained infertility and good quality oocytes.
Background Sperm abnormalities may negatively affect embryo development. Objectives To determine the influence of sperm abnormalities (morphology, motility, DNA fragmentation) on embryo morphokinetic variables and clinical outcome of conventional IVF. Materials and methods Participants were 86 couples undergoing in vitro fertilization (IVF). Sperm morphology was evaluated according to the strict criteria proposed by Kruger/Tygerberg. CASA system was applied for sperm motility assessment. Sperm DNA fragmentation was assessed by the chromatin structure assay (SCSA). Morphokinetic parameters were determined in 223 embryos obtained from conventional IVF only and cultured in a single‐step medium using time‐lapse imaging technology. Results Time‐lapse variables from the initial embryo development, such as time of pronuclei fading (tPNf) and time for two cells (t2), were those more strongly related with abnormalities of sperm motility, morphology, and DNA fragmentation. Sperm morphological abnormalities rather than sperm motility were more closely associated with embryo morphokinetics. Sperm head defects were mainly correlated with the last stages of embryonic development (t9 to tHB), sperm midpiece defects with intermediate cleaving embryos (t5‐t9), and sperm tail defects with the initial stages of embryonic development (tPNa‐t4). Excess residual cytoplasm was positively correlated with all embryo morphokinetic parameters except t2 and tM. Absence of acrosomes, pinheads, coiled tails, and multiple sperm morphological defects correlated negatively with time‐lapse embryo morphokinetic variables. Discussion A large number of sperm‐related variables, including frequency of specific morphological defects, morphological indexes, DNA fragmentation and motility, and time‐lapse embryo variables, such as time intervals based mainly of 15 time points were recorded. Conclusion There were strong associations between specific sperm defects of the head, midpiece, and tail with certain stages of embryonic development from observation of pronuclei to the hatched blastocyst. Coiled tail, cumulative head defects, and multiple abnormalities index (MAI) were associated both with embryo morphokinetics and the implantation success.
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