The effect of the assisted zona hatching (AZH) procedure was investigated on 135 cycles with a poor prognosis of pregnancy due to: (i) maternal age > or = 38 years (45 cycles); (ii) three or more failed in-vitro fertilization (IVF) attempts (70 cycles), and (iii) patients possessing both inclusion criteria (20 cycles). The control groups (113 cycles) included patients possessing the same characteristics (42, 53 and 18 cycles respectively) and who did not undergo the AZH procedure. A total of 505 embryos was treated with AZH before transfer, resulting in: 14, 25 and 6 clinical pregnancies. The percentage of clinical pregnancies per cycle was significantly higher than controls for the first (31 vs 10% in control 1, P < 0.05) and second groups (36 vs 17% in control 2, P < 0.05). No significant difference in percentage of clinical pregnancies was found for the third group (30 vs 6%). Similarly, higher rates of implantation were obtained (11.5, 15 and 11%) compared to the respective controls (4%, P < 0.02; 6.3%, P < 0.01; and 1.5%). The rate of miscarriage in the AZH groups was similar to that obtained in the controls (22 vs 21%). Finally, the morphological analysis of the embryos transferred revealed that the poor prognosis condition is associated to a significantly slower rate of development and a higher rate of fragmentation. The present results indicate that AZH procedure improves pregnancy and implantation rates in patients with a poor prognosis of pregnancy by facilitating the hatching process in embryos which would otherwise be trapped inside the zona pellucida.
Purpose In this work, we describe a system for the morphological scoring of human oocytes prior to fertilisation and use this system to test whether oocyte morphology is an indicator of fertilisation, embryo development and implantation potential. Methods The study is a prospective trial of the use of oocyte morphological scores in 822 patients undergoing their first cycle of ICSI. Analyses of oocytes were performed prior to ICSI procedures and the scores compared with fertilisation rates, embryo quality and clinical results.Results 'Top quality' oocytes had a significantly higher level of fertilisation (96%) as compared to low scoring oocytes (25.6%). Where top quality oocytes formed top quality embryos, we noted a clinical success rate of 63.4%. Conclusions Clinical success rates were increased in cases where top quality oocytes formed top quality embryos after ICSI. The analysis of oocyte morphology may represent a positive selection feature during ICSI.
A reduced time interval of oocyte exposure to spermatozoa was investigated to assess whether it could enhance oocyte development and improve embryo viability, especially in cases of male factor infertility. A total of 167 patients were included in a prospective randomized study. They were randomly allocated to two major study groups, A (n = 85) and B (control group; n = 82). The oocytes from group A patients were exposed to spermatozoa for only 1 h; those from group B were exposed for 16 h. The two study groups were then subdivided according to semen quality for further analysis of the results. Significantly higher percentages were obtained in group A than in group B in terms of the fertilization rate (74 versus 68%, P < 0.025), cleavage rate (53 versus 41%, P < 0.005), pregnancy rate (27 versus 12%, P < 0.05) and implantation rate (11 versus 6%, P < 0.05). In addition, an increased fertilization rate was achieved in oocytes exposed to male factor spermatozoa for only 1 h compared with the conventional incubation period (78 versus 65%, P < 0.01). Advanced cellular stages (55 versus 41%, P < 0.02) and higher implantation rates (13 versus 4%, P < 0.05) were attained in the subgroup whose oocytes were exposed to normal spermatozoa for 1 h compared with the male factor spermatozoa with the standard culture interval. The higher fertilization rates, enhanced embryo development and viability achieved in group A indicate that prolonged exposure of oocytes to high concentrations of spermatozoa is detrimental, decreasing sperm-oocyte interaction and subsequent embryo implantation, particularly in male factor patients.
PGD (preimplantation genetic diagnosis) of aneuploidy for chromosomes X, Y, 13, 18 and 21 was carried out on 196 embryos from 36 infertile patients classified with a poor prognosis due to (i) maternal age, (ii) repeated in-vitro fertilization (IVF) failures and (iii) mosaic karyotype. The percentage of abnormal embryos was comparable in the three groups of patients: maternal age 63%, repeated IVF failure 57%, and mosaic karyotype 62%. The analysis of the overall data revealed an increased incidence of abnormal embryos in the older age categories (predominantly due to aneuploidy), even in embryos at the 7- to 8-cell stage. In addition, the percentage of chromosomally abnormal embryos was directly proportional to the number of IVF failures, where the increase in chromosomal abnormalities was not correlated to aneuploidy but to other aberrations such as mosaicism and polyploidy. Following PGD, 28 patients had at least one embryo transferred that appeared normal by fluorescent in-situ hybridization (FISH). Four clinical pregnancies resulted, with an implantation rate of 10% per normal embryo. In conclusion, the high rate of chromosomally abnormal embryos in poor prognosis patients may have been the cause of implantation failure in their previous IVF cycles. Therefore, the possibility of transferring embryos with a normal FISH complement could improve the chance of pregnancy in this category of patients.
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