Expression of glycodelin and cyclooxygenase-2 in human endometrial tissue following three-dimensional culture.

PROBLEM: Our previous study showed that in vitro culture of human endometrial tissue in a three-dimensional (3D) fibrin matrix could mimic the early stages of endometriosis with invasion, gland and stroma formation and sprouting of new vessels. The objective of the present study was to evaluate the expression of glycodelin (Gd) and cyclooxygenase-2 (COX-2), two angiogenic factors, to further validate the 3D culture model of endometriosis. METHOD OF STUDY: Human endometrial fragments were obtained from endometrial biopsies and placed in a 3D fibrin matrix culture. Immunohistochemistry with specific antibodies to Gd and COX-2 was used to examine endometrial epithelium and blood vessels, and 4, 6-diamidino-2-phenylindole staining was used for nuclear identification. RESULTS: Three-dimensional culture of human endometrial tissue in the fibrin matrix resulted in the proliferation of endometrial stromal cells, glandular epithelium and angiogenesis. Gd positive glandular epithelium was seen in 85% of wells with developing endometrial glands and COX-2 positive new vessels were seen in 80% of wells with angiogenesis-like structures after 4 weeks of culture. CONCLUSION: Our findings confirm that angiogenesis occurs following the culture of endometrial tissue in the 3D fibrin matrix, and suggests that Gd and COX-2 might play important roles in promoting neovascularization and cell proliferation in the establishment of endometriosis.

Complete failed fertilization after intracytoplasmic sperm injection--analysis of 10 years' data.

OBJECTIVE: To investigate incidence and causes of complete failed fertilization after intracytoplasmic sperm injection (ICSI) in a tertiary care facility. METHODS: A total of 1,779 cycles between February 1994 and December 2003 were analyzed. Study parameters were female age, infertility diagnosis, ovarian stimulation protocol, estradiol level on day of hCG administration, number of follicles, number of oocytes retrieved, number of oocytes injected, and semen parameters. RESULTS: Complete failed fertilization occurred in 23 cycles (1.29%) involving a total of 85 oocytes injected. Infertility causes among patients with failed fertilization included unexplained (43.6%), male factor (26%), presence of more than one factor (17.4%), hysterectomy (4.4%), premature ovarian failure (4.3%), and advanced age (4.3%). In 12 cycles (52%), fewer than 5 follicles were present. In three (13%) cycles, no mature (MII) oocyte was available and in 61% (14/23) fewer than 3 MII oocytes were available for ICSI. Immotile sperm was used for ICSI in 5 cycles (21.7%). The source of sperm in 17 (74%) cycles was from ejaculate, in 4 cycles from testicular aspiration (TESA), one from percutaneous epididymal sperm aspiration (PESA) and one from retrograde ejaculation. CONCLUSIONS: Our data indicate that major contributing factors to failed fertilization after intracytoplasmic sperm injection are number of MII oocytes retrieved and availability of viable sperm for injection. Although the incidence of complete failed fertilization is not remarkable, it may increase with increasing patient age and a lower number of follicles.

Duration of sexual arousal predicts semen parameters for masturbatory ejaculates.

A man's semen parameters may vary considerably from one specimen to the next, partly due to variability in the conditions under which the specimens are produced. In the present study, the relationship between the duration of preejaculatory sexual arousal and the quality of semen produced by masturbation was investigated. Twenty-five regular semen donors aged 22-44 provided a total of 292 semen specimens (median 11 per donor) over a period of 4 months. Each specimen was produced after a minimum of 3 days of ejaculatory abstinence and measures included the time taken to produce the specimen, ejaculate volume, sperm concentration, and percent motility. Linear regression revealed that, controlling for donor identity, there was a significant (t=2.13, P<.05) positive relationship between the time taken to produce a specimen and sperm concentration. We conclude that the duration of preejaculatory sexual arousal is an important predictor of ejaculate quality for specimens produced by masturbation and that variation in the duration of preejaculatory arousal may contribute to within-male fluctuations in semen parameters over time.