1 Centre For Reproductive Medicine and 2 Center for Medical Genetics, University Hospital, Dutch-speaking Brussels Free University (Vrije Universiteit Brussel), Laarbeeklaan 101, B-1090 Brussels, Belgium
3 To whom correspondence should be addressed. e-mail: valerie.vernaeve{at}az.vub.ac.be
![]() |
Abstract |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Key words: azoospermia/follow-up/ICSI/pregnancy outcome/testicular sperm
![]() |
Introduction |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
The different percentages found in the published studies about major and minor congenital malformations cannot be compared, but overall the data in large and reliable surveys do not indicate a higher rate of malformations in ICSI children than in IVF- or naturally conceived children (Wennerholm et al., 2000a; Bonduelle et al., 2002a
). However, the study by Ericson and Källen (2001
) found an increased risk of the total malformation rate after IVF which could mainly be explained by maternal factors. Furthermore, the limited available data on ICSI fetal karyotypes reveal that, in comparison with a general neonatal population, there is a slight but increased risk of chromosomal anomalies, predominantly affecting the sex chromosomes (Bonduelle et al., 2002b
).
Today, ICSI is also widely used for patients with azoospermia even when severe testicular failure is present. Since there is increasing evidence that in these patients spermatozoa have an increased chromosomal aneuploidy rate (Bernardini et al., 2000; Martin et al., 2000
; Levron et al., 2001
; Mateizel et al., 2002
), follow-up of the pregnancies obtained after the use of testicular sperm from these patients is very important. Few studies involving a small number of patients have been published on the outcome of pregnancies and the health of children born after ICSI with testicular sperm (Aytoz et al., 1998
; Wennerholm et al., 2000b
; Bonduelle et al., 2002a
; Ludwig et al., 2002
). Furthermore, a distinction between normal spermatogenesis and spermatogenetic failure is lacking.
The aim of this study is to analyse the pregnancy outcome and neonatal outcome of children born after ICSI with testicular sperm in patients with obstructive and non-obstructive azoospermia.
![]() |
Materials and methods |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Pregnancies obtained after ICSI with both fresh and frozen testicular spermatozoa were included in the study.
Technical procedures, including testicular sperm recovery, ovarian stimulation protocols, micro-injection procedure, embryo culture and transfer, and luteal phase support, have been described previously (Verheyen et al., 1995; Tournaye et al., 1997
; Joris et al., 1998
; Van Steirteghem et al., 1998
).
A rise in serum HCG on two consecutive occasions from 11 days after transfer indicated pregnancy. Each pregnancy with at least one sac revealed by ultrasonography 5 weeks after transfer was considered as a clinical pregnancy. If there was no sac, the pregnancy was considered subclinical. Gestational age was calculated as the time between the beginning of the last menstrual period and the date of birth of the child. The last menstrual period was calculated by adding 15 days to the date of embryo transfer.
We evaluated both early (before 20 weeks of gestation) and late (20 weeks of gestation) pregnancy outcome in the NOA and obstuctive azoospermia (OA) cohorts.
Abortion was defined as pregnancy loss before 20 weeks of gestational age, and preterm delivery was defined as delivery of a live born or stillborn infant before 37 weeks of gestational age. A live born or stillborn infant weighing <2500 g at birth was considered a low birth weight infant. A live born or stillborn infant weighing <1500 g at birth was considered a very low birth weight infant. The death of a fetus of at least 20 weeks gestation before delivery was defined as intrauterine death, and the death of an infant during the first week following delivery was defined as early neonatal death. The early perinatal mortality rate was expressed as the sum of stillbirths and early neonatal deaths per 1000 births.
Major malformation was defined as those malformations causing death or functional impairment, or requiring surgical correction. The remaining malformations were considered minor malformations.
Since all outcome measures were not available for all patients, calculations were performed on appropriate data subsets.
Comparisons of the NOA and OA groups for qualitative variables were performed using a Fisher exact test. A MannWhitney test was used when data were not normally distributed. A P-value of <0.05 was considered to be statistically significant. Additional relative risk (RR) with corresponding 95% confidence interval (CI) was calculated whenever relevant.
This study was approved by our institutional review board.
![]() |
Results |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
The mean age of the women was 31.4 years (95% CI, 29.733.0) versus 32.7 years (95% CI, 31.833.6) in the NOA and OA group, respectively. Primigravidae were more common in the NOA group (51 out of 67 with known gestational status versus 127 out of 203 with known status, i.e. 76 versus 63%), but statistical significance was not reached (P = 0.053). The parity was not statistically different between the two cohorts.
There were no statistical significant differences with respect to the outcome of the pregnancies in the two groups studied (Table I).
|
Of the 196 children in the OA group, 193 were live born and three were stillborn. One-hundred and ten children were from singleton pregnancies, 74 were from twin pregnancies and 12 were from triplet pregnancies.
There was a strong tendency towards a lower gestational age among the singletons and a higher percentage of preterm twins in the NOA group. The gestational age at birth and percentages of preterm deliveries are summarized in Table II
|
Three fetuses died in utero after 20 weeks in both groups, and one infant died in the early neonatal period in the NOA group. The early perinatal mortality rate was thus 66 per 1000 births (n = 4) and 15 per 1000 (n = 3) in the NOA and OA group, respectively (RR: 4.3, 95% CI 0.823.7).
Major malformations were present in 4% of the liveborn children (n = 2 out of 54 with a known result) obtained with testicular sperm of NOA men versus in 3% in children of OA men (n = 5 out of 188 with a known result) (RR: 1.4, 95% CI 0.197.8). One polymalformative syndrome was present in a stillborn child of the NOA group. The minor malformation rate in live born children in the NOA and OA group was 2% (n = 1 out of 54 with a known result) versus 4% (n = 8 out of 188 with a known result) (RR: 0.4, 95% CI 0.023.27) (Table III).
|
![]() |
Discussion |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Few publications so far have addressed the obstetric and neonatal outcome of children born after ICSI using testicular sperm. The study by Ludwig et al. (2002) analysed the outcome of 229 pregnancies in which testicular sperm was used and found no additional risk of major malformations in children born after the use of testicular spermatozoa. The major malformation rate, up to 8 weeks after birth, in this study was 9% based on live born and stillborn children and including spontaneous and induced abortions, compared wth 8.4% in ICSI with ejaculated spermatozoa. Bonduelle et al. (2002a
) examined the outcome of malformation rates and found no differences, up to 8 weeks after birth, between ejaculated (3.4%, n = 2477) and testicular sperm (2.9%, n = 206). The study performed by Wennerholm et al. (2000b
) found no major malformations in the 31 children born after the use of testicular sperm for ICSI. In these studies, the subgroups of testicular sperm are small and no discrimination is made between OA and NOA. Only the study by Palermo et al. (1999
) differentiated between these two subgroups, although it was not clear whether it was done on a clinical or a histopathological basis. In the 34 pregnancies obtained after the use of testicular sperm, eight were classified as obstructive and 26 as non-obstructive. Similarly, in this study, the prevalence of congenital malformation did not vary in relation to the sample origin (or the cause of azoospemia). In our study, we found a malformation rate of 4% after the use of testicular sperm of NOA patients and 3% after the use of testicular sperm of OA patients. These rates are comparable with the rates observed in the study by Bonduelle et al. (2000a
) where a 3.4% major malformation rate was found in ICSI children after the use of ejaculated sperm, using the same methodology and definitions as in this study.
With regards to the pregnancy outcome, we observed a strong tendency towards lower gestational age among the singletons and a higher percentage of premature twins in the NOA group, when comparing two different subgroups of azoospermic patients.
In this study we did not include a historical control group such as oligoasthenoteratozoospermic patients or spontaneous conceptions. The sample size of our current data set would be too small to draw any valid conclusion. However, given our findings and the concerns about the use of immature testicular spermatozoa from men with spermatogenetic failure, further study is recommended. In view of the low pregnancy rate after ICSI in NOA, a multicentre study in which a distinction is made between azoospermic patients with OA and those with NOA is suggested.
![]() |
Acknowledgements |
---|
![]() |
References |
---|
![]() ![]() ![]() ![]() ![]() ![]() ![]() |
---|
Bernardini, L., Gianaroli, L., Fortini, D., Conte, N., Magli, C., Cavani, S., Gaggero, G., Tindiglia, C., Ragni, N. and Venturini, P.L. (2000) Frequency of hyper-, hypohaploidy and diploidy in ejaculated, epididymal and testicular germ cells of infertile patients. Hum. Reprod., 15, 21652172.
Bonduelle, M., Liebaers, I., Deketelaere, V., Derde, M.P., Camus, M., Devroey, P. and Van Steirteghem, A. (2002a) Neonatal data on a cohort of 2889 infants born after ICSI (19911999) and of 2995 infants born after IVF (19831999). Hum. Reprod., 17, 671694.
Bonduelle, M., Van Assche, E., Joris, H., Keymolen, K., Devroey, P., Van Steirteghem. A. and Liebaers, I. (2002b) Prenatal testing in ICSI pregnancies: incidence of chromosomal anomalies in 1586 karyotypes and relation to sperm parameters. Hum. Reprod., 17, 26002614.
Ericson, A. and Källen, B. (2001) Congenital malformations in infants born after IVF: a population-based study. Hum. Reprod., 16, 504509.
Hewitson, L., Simerly, C. and Schatten, G. (2002) Fate of sperm components during assisted reproduction: implication for infertility. Hum. Fertil., 5, 111116.
Joris, H., Nagy, Z., Van de Velde, H., De Vos, A. and Van Steirteghem, A. (1998) Intracytoplasmic sperm injection: laboratory set-up and injection procedure. Hum. Reprod., 13 (Suppl. 1), 7686.
Levron, J., Aviram-Goldring, A., Madgar, I., Raviv, G., Barkai, G. and Dor, J. (2001) Sperm chromosome abnormalities in men with severe male factor infertility who are undergoing in vitro fertilization with intracytoplasmic sperm injection. Fertil. Steril., 76, 479484.[CrossRef][ISI][Medline]
Ludwig, M. and Katalinic, A. (2002) Malformation rate in fetuses and children conceived after ICSI: results of a prospective cohort study. Reprod. Biomed. Online, 5, 171178.[Medline]
Martin, R.H., Greene, C., Rademaker, A., Barclay, L., Ko, E. and Chernos, J. (2000) Chromosome analysis of spermatozoa extracted from testes of men with non-obstructive azoospermia. Hum. Reprod., 15, 11211124.
Mateizel, I., Verheyen, G., Van Assche, E., Tournaye, H., Liebaers, I. and Van Steirteghem, A. (2002) FISH analysis of chromosome X, Y and 18 abnormalities in testicular sperm from azoospermic patients. Hum. Reprod., 17, 22492257.
Palermo, G., Joris, H., Devroey, P. and Van Steirteghem, A.C. (1992) Pregnancies after intracytoplasmic injection of single spermatozoon into an oocyte. Lancet, 340, 1718.[ISI][Medline]
Palermo, G., Schlegel, P.N., Hariprashad, J.J., Ergun, B., Mielnik, A., Zaninovic, N., Veeck, L.L. and Rosenwaks, Z. (1999) Fertilization and pregnancy outcome with intracytoplasmic sperm injection for azoospermic men. Hum. Reprod., 14, 741748.
Tesarik, J. and Mendoza, C. (1996) Genomic imprinting abnormalities: a new potential risk of assisted reproduction. Mol. Hum. Reprod., 2, 295298.[Medline]
Tournaye, H., Camus, M., Vandervorst, M., Nagy, Z., Joris, H., Van Steirteghem, A. et al. (1997) Surgical sperm retrieval for intracytoplasmic sperm injection. Int. J. Androl., 20 (Suppl. 3), 6973.[ISI][Medline]
VanSteirteghem, A., Nagy, P., Joris, H., Janssenswillen, C., Staessen, C., Verheyen, G., Camus, M., Tournaye, H. and Devroey, P. (1998) Results of intracytoplasmic sperm injection with ejaculated, fresh and frozen-thawed epididymal and testicular spermatozoa. Hum. Reprod., 13 (Suppl. 1), 13442.[ISI][Medline]
Verheyen, G., De Croo, I., Tournaye, H., Pletincx, I., Devroey, P. and Van Steirteghem, A.C. (1995) Comparison of four mechanical methods to retrieve spermatozoa from testicular tissue. Hum. Reprod., 10, 29562959.[Abstract]
Vernaeve, V., Tournaye, H., Osmanagaoglu, K., Verheyen, G., Van Steirteghem, A. and Devroey, P. (2003) Intracytoplasmic sperm injection with testicular spermatozoa is less successful in men with nonobstructive azoospermia than in men with obstructive azoospermia. Fertil. Steril., 79, 529533.[CrossRef][ISI][Medline]
Wennerholm, U.-B., Bergh, C., Hamberger, L., Lundin, K., Nilsson, L., Wikland, M. and Källén, B. (2000a) Incidence of congenital malformations in children born after ICSI. Hum. Reprod., 15, 944948.
Wennerholm, U.-B., Bergh, C., Hamberger, L., Westlander, G., Wikland, M. and Wood, M. (2000b) Obstetric outcome of pregnancies following ICSI, classified according to sperm origin and quality. Hum. Reprod., 15, 11891194.
Submitted on November 16, 2002; resubmitted on May 12, 2003; accepted on June 18, 2003.