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Intrauterine insemination: Which sperm characteristics matter?
tility-Ctr de Fertilizac¸ ao Assistida, Sao Paulo, Brazil; UNICAMP, Campinas, Brazil. Objectives: Using cryptorchidic surgical induced rats, we assessed the suppression of spermatogenesis and the results obtained from TESE (testicular sperm extraction) and TESA (testicular sperm aspiration). Design: Experimental study. Material and Methods: Forty adult rats (Wistar) were used in the study. First of all, 8 rats underwent a bilateral induced cryptorquidia to induce spermatogenesis disorders in different time interval (2 rats/7 days of cryptorchidism; 2 rats/10 days; 2 rats/15 days and 2 rats/30 days). After histological analysis in each interval, a 15-day period was chosen. From the remaining animals 25 underwent a bilateral induced cryptorquidia and 7 were used as sham-operated control. Tissue samples were obtained under stereoscopic microscopy from both testicles in the control group and 15 days after cryptorchidia induced. Firstly, all 32 right testicles underwent a TESA procedure through two points of needle entrance, but searching for spermatozoa in all testis extension, and afterwards TESE in 3 different poles of the testicles (superior, middle, and inferior). In the same way, 32 left testis underwent a TESA procedure in order to search for spermatozoa in 3 different poles (superior, middle, and inferior), and afterwards, TESE in a similar way described for the left testis. Samples were processed according to usual procedure for TESE and TESA in humans. According to the number of spermatozoon retrieved from the testis, 4 scores were created: 0 (absent, no sperm), 1 (few sperm, 1 to 3 x 106 sperm/ml), 2 (fairly, 3 to 15 x 106 sperm/ml), and 3 (good, more than 3 x 106 sperm/ml and lots of sperm outside the reticule grid of the Makler chamber). Afterwards, testicles were extracted, weighted and a morphological analysis at conventional light microscopy level was done. Results: The weight of the left testis in the cryptorchidic rats was significantly lower compared to the left testis of the control rats (p⫽0.0002). Although no significant different, a trend was seen in the right testis of the cryptorchidic rats to have lower weight compared to the control rats (p⫽0.09). A histological study of the testicles showed that experimental cryptorchidism was induced in adults rats and caused a severe failure in a spermatogenesis process. Significant differences were seen between the cryptorchidic and control rats in the scores (p⫽0.012). When we compared the scores seen in the left and right testis employing the TESA technique, no differences were observed (p⫽ 0.58). Also, there were no differences in the scores obtained from the left and right testis with the TESE technique. Comparing the TESE and the TESA, no differences were seen in the number of spermatozoa retrieved (p⬎0.05). No differences were seen in the superior, middle and inferior poles of the testis, irrespective the type of biopsy technique employed. Conclusions: It seems that 15-day period cryptorchidia is enough to induce spermatogenesis disorders. No differences were seen in the scores obtained in the right and left testicles, irrespective the testicular pole. Furthermore and more important, no differences in the score levels were seen in the two techniques.
P-262 Intrauterine insemination: Which sperm characteristics matter? David T. Wyncott, Nikos Vlahos, Jairo Garcia, Howard Zacur, Edward Wallach, Yulian Zhao. Johns Hopkins Hosp, Baltimore, MD. Objective: To evaluate how sperm characteristics including sperm concentration, motility, forward progression and total motile sperm count influence the outcome of infertility treatment using intrauterine insemination (IUI). Design: Retrospective cohort study. Materials and Methods: One thousand and seven IUI treatment cycles from June 1999 to Oct 2002 were included in the study. The age (Mean⫾SD) for the male patients was 37.1⫾13.3 years and for the female patients was 35.2⫾4.5 years. Sperm parameters before and after preparation by density gradient separation were evaluated according to World Health Organization (WHO) guidelines. Statistical analysis for sperm characteristics in relation to pregnancy outcome was performed using two-tailed t-test, Pearson correlation, and Chi-square where appropriate. A p⬍.05 was considered statistically significant. Results: Clinical pregnancy occurred in 121 (12%) of 1007 cycles. Comparison of semen parameters in pregnancy versus non-pregnancy cycles is shown in the accompanying table.
FERTILITY & STERILITY威
TMS ⫽ total motile sperm, a progression 4-1⫽A-D (WHO), * ⫽ statistically significant Pregnancy was positively correlated with initial sperm motility (p⬍0.05) and processed forward progression (p⬍0.05). No pregnancies occurred when the motility was ⬍26% and processed forward progression was ⬍3. When between-group comparisons were performed, sperm motility of the initial semen specimen ⬎80% (p⫽0.02) offered a favorable pregnancy rate (PR) at 17.6%. After processing for insemination, sperm concentration of 51-100 x 106/ml (p⫽0.01, PR 16.5%), TMS of 11-100 x 106 (p⫽0.049, PR 13.5%) and forward progression of ⫽3 (p⫽0.02, PR 12.8%) are optimal for pregnancy. Initial sperm concentration, forward progression and total motile sperm count had no significant effect on IUI outcome. Conclusions: This study demonstrated that sperm motility is an independent factor predicting IUI pregnancy. Forward progression equal to or greater than 3 in processed specimen may be necessary for a positive IUI outcome. Sperm concentration of 51-100 x 106/ml and TMS ⫽11 million per insemination offer the best potential for success.
P-263 Blastocyst formation rate after ICSI with extreme oligozoospermic samples. Anick De Vos, Lisbet Van Landuyt, Kristien De Bent, Hubert Joris, Paul Devroey, Andre´ Van Steirteghem. Ctr for Reproductive Medicine, Brussels, Belgium. Objective: Male factor infertility is believed to have an impact not only on fertilisation rate but also on the early stages of human embryonic development. Several studies support a paternal influence on blastocyst formation when comparing ICSI and IVF cycles: the blastocyst formation rate being consistently lower in ICSI cycles than in IVF cycles. In the present study the blastocyst formation rate using extreme oligozoospermic samples for ICSI was evaluated. Design: Retrospective case-controlled study. Materials and Methods: The study group involved 115 ICSI cycles where the sperm count was less than 0.1 million per ml. The control group consisted of 115 ICSI cycles from the same period (January 2001-July 2002) where the sperm count was at least 20 million per ml. They were matched for female age, indication, rank of trial and number of cumulusoocyte-complexes. The outcome parameters were 2-PN fertilisation rate, number of four-cell (type A and B) and eight-cell (type A, B and C) embryos on day 2 and day 3 respectively, compaction on day 4, and good quality blastocyst formation (at least stage 3 and score BB for ICM and trophectoderm) on day 5 and day 6. Results: Evaluating the individual sperm morphology at the moment of injection revealed that more abnormal forms were injected in the study group (13.6%) than in the control group (1.2%; P⬍0.001). The fertilisation rate was significantly different between the two groups (P⬍0.001). Significant lower percentages of eight-cell and compact embryos were obtained in the study group (P⬍0.05). However, the percentages of good quality blastocysts on day 5 and day 6 were not different between the two groups.
Conclusion: Less eight-cell and compact embryos are available after ICSI in cases of extreme oligozoospermia. These results point to a paternal effect on human embryonic development that becomes evident as early as the activation of the embryonic genome. However, further development to the blastocyst stage seems not to be impaired.
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P-262 Intrauterine insemination: Which sperm characteristics matter? David T. Wyncott, Nikos Vlahos, Jairo Garcia, Howard Zacur, Edward Wallach, Yulian Zhao. Johns Hopkins Hosp, Baltimore, MD. Objective: To evaluate how sperm characteristics including sperm concentration, motility, forward progression and total motile sperm count influence the outcome of infertility treatment using intrauterine insemination (IUI). Design: Retrospective cohort study. Materials and Methods: One thousand and seven IUI treatment cycles from June 1999 to Oct 2002 were included in the study. The age (Mean⫾SD) for the male patients was 37.1⫾13.3 years and for the female patients was 35.2⫾4.5 years. Sperm parameters before and after preparation by density gradient separation were evaluated according to World Health Organization (WHO) guidelines. Statistical analysis for sperm characteristics in relation to pregnancy outcome was performed using two-tailed t-test, Pearson correlation, and Chi-square where appropriate. A p⬍.05 was considered statistically significant. Results: Clinical pregnancy occurred in 121 (12%) of 1007 cycles. Comparison of semen parameters in pregnancy versus non-pregnancy cycles is shown in the accompanying table.
FERTILITY & STERILITY威
TMS ⫽ total motile sperm, a progression 4-1⫽A-D (WHO), * ⫽ statistically significant Pregnancy was positively correlated with initial sperm motility (p⬍0.05) and processed forward progression (p⬍0.05). No pregnancies occurred when the motility was ⬍26% and processed forward progression was ⬍3. When between-group comparisons were performed, sperm motility of the initial semen specimen ⬎80% (p⫽0.02) offered a favorable pregnancy rate (PR) at 17.6%. After processing for insemination, sperm concentration of 51-100 x 106/ml (p⫽0.01, PR 16.5%), TMS of 11-100 x 106 (p⫽0.049, PR 13.5%) and forward progression of ⫽3 (p⫽0.02, PR 12.8%) are optimal for pregnancy. Initial sperm concentration, forward progression and total motile sperm count had no significant effect on IUI outcome. Conclusions: This study demonstrated that sperm motility is an independent factor predicting IUI pregnancy. Forward progression equal to or greater than 3 in processed specimen may be necessary for a positive IUI outcome. Sperm concentration of 51-100 x 106/ml and TMS ⫽11 million per insemination offer the best potential for success.
P-263 Blastocyst formation rate after ICSI with extreme oligozoospermic samples. Anick De Vos, Lisbet Van Landuyt, Kristien De Bent, Hubert Joris, Paul Devroey, Andre´ Van Steirteghem. Ctr for Reproductive Medicine, Brussels, Belgium. Objective: Male factor infertility is believed to have an impact not only on fertilisation rate but also on the early stages of human embryonic development. Several studies support a paternal influence on blastocyst formation when comparing ICSI and IVF cycles: the blastocyst formation rate being consistently lower in ICSI cycles than in IVF cycles. In the present study the blastocyst formation rate using extreme oligozoospermic samples for ICSI was evaluated. Design: Retrospective case-controlled study. Materials and Methods: The study group involved 115 ICSI cycles where the sperm count was less than 0.1 million per ml. The control group consisted of 115 ICSI cycles from the same period (January 2001-July 2002) where the sperm count was at least 20 million per ml. They were matched for female age, indication, rank of trial and number of cumulusoocyte-complexes. The outcome parameters were 2-PN fertilisation rate, number of four-cell (type A and B) and eight-cell (type A, B and C) embryos on day 2 and day 3 respectively, compaction on day 4, and good quality blastocyst formation (at least stage 3 and score BB for ICM and trophectoderm) on day 5 and day 6. Results: Evaluating the individual sperm morphology at the moment of injection revealed that more abnormal forms were injected in the study group (13.6%) than in the control group (1.2%; P⬍0.001). The fertilisation rate was significantly different between the two groups (P⬍0.001). Significant lower percentages of eight-cell and compact embryos were obtained in the study group (P⬍0.05). However, the percentages of good quality blastocysts on day 5 and day 6 were not different between the two groups.
Conclusion: Less eight-cell and compact embryos are available after ICSI in cases of extreme oligozoospermia. These results point to a paternal effect on human embryonic development that becomes evident as early as the activation of the embryonic genome. However, further development to the blastocyst stage seems not to be impaired.
S209