kızlar makaleyi gönderiyorum Rescue of spermatogenesis arrest in azoospermic
men after long-term gonadotropin treatment
Several studies have shown that FSH treatment can improve sperm production quantitatively and increase the
spermatogonial population in oligozoospermic men with normal hormonal profiles. In this study, we describe the
results of long-term gonadotropin therapy of normogonadotropic patients with nonobstructive azoospermia. (Fertil
Steril 2006;86:466–8. ©2006 by American Society for Reproductive Medicine.)
Several forms of male-factor infertility have been treated
successfully with intracytoplasmic injection (ICSI), even
for patients who have few spermatozoa in their ejaculate or
in samples extracted from testicular biopsies. However, a
number of azoospermic men may fail to have any sperm in
their testicular biopsies because of complete maturation
arrest.
Follicle-stimulating hormone plays a key role in spermatogenesis
and sperm production (1, 2). Gonadotropin
treatment for men with azoospermia that is caused by hypogonadotropic
hypogonadism has been well documented (3, 4).
Several studies have shown that FSH treatment can improve
sperm production quantitatively and increase the spermatogonial
population in oligozoospermic men with normal hormonal
profiles (5–7). However, the effectiveness of gonadotropin
treatment on normogonadotropic azoospermic patients
is debatable. In this report, we describe the results of longterm
gonadotropin therapy on normogonadotropic patients
with nonobstructive azoospermia.
A total of 49 infertile men (aged 32–41 years) with
nonobstructive azoospermia were included in this clinical
study in two different IVF centers; of the men, 23 were
treated in the first center, and 26, in the second. All of the
patients had normal hormonal profiles and normal testicular
volume, and none had any history of varicocele or cryptorchidis.
In addition, genetic analysis confirmed the absence
of Y-chromosome microdeletion in all the patients.
Semen analysis was performed at both centers at 2-week
intervals. To discover or rule out the presence of sperm, the
ejaculate was diluted with equal volumes of media and
centrifuged at 1,000g for 10 minutes. The supernatant was
removed, and the whole pellet was diluted in a maximum
of 50 L and thoroughly examined for the presence of
sperm. All of the patients were shown to have a complete
absence of spermatozoa in the ejaculate.
Explorative open multiple testicular biopsies were performed,
and tissues were dissected and digested with collagenase
and DNAse (Sigma, Milan, Italy). Examination of
all testicular samples failed to produce any mature sperm in
all samples. Histological examination of the testicular biopsies
showed maturation arrest of spermatogenesis at the
spermatocytes or spermatids stage. Both clinical IVF centers
used the identical protocol in performing testicular
biopsies and processing testicular samples. The procedure
was explained to the patients, and the patients gave informed
consent before treatment. The procedure was approved
by the local institutional ethics committee.
All patients underwent long-term gonadotropin therapy
by SC administration of 75 IU of recombinant FSH (rFSH;
Gonal-F, Serono, Rome, Italy) on alternate days for the first
2 months.
At the beginning of the 3rd month, the rFSH dose was
increased to 150 IU on alternate days, and from the 4th
month onward, 2,000 IU of hCG (Gonasi, IBSA, Switzerland)
were administered twice weekly in addition to the
rFSH. The same treatment protocol was followed at both
clinical centers. After 6 months of gonadotropin therapy,
analysis of the fresh semen samples failed to detect spermatozoa.
Multiple testicular biopsies then were performed
as a potentially more reliable means to detect the presence
of sperm, If sperm were present, the biopsy material was
cryopreserved, and the female partners were enrolled for
IVF treatment.
Female partners, aged 30 to 38 years, underwent a standard
down-regulation with GnRH analogue (Decapeptyl;
Ipsen, Milan, Italy) and rFSH (Gonal-F; Serono, Rome,
Italy) for ovarian stimulation. Retrieved mature oocytes
were injected by using sperm found and extracted from the
frozen–thawed testicular biopsies. Fertilization was assessed,
and normally fertilized oocytes were cultured for a
further 24 hours. Embryo transfer took place 48 hours after
insemination.
After the gonadotropin treatment, few sperm were found
in the processed testicular biopsies in 11 of 49 patients
(22.4%
. The total sperm number ranged between 0.001 to
0.004 106, with only one third of the total spermatozoa
having in situ motility. Fifty-three mature oocytes were
Received August 28, 2005; revised and accepted December 20, 2005.
Reprint requests: Helmy Selman, Ph.D., Spatocco Hospital, Viale
Amendola 93, 66100 Chieti, Italy (FAX: 39-075-578-3260; E-mail:
selmanha@yahoo.com).
466 Fertility and Sterility Vol. 86, No. 2, August 2006 0015-0282/06/$32.00
Copyright ©2006 American Society for Reproductive Medicine, Published by Elsevier Inc. doi:10.1016/j.fertnstert.2005.12.055injected with the sperm that was extracted from frozen–
thawed testicular biopsies, resulting in 34 normally fertilized
oocytes (64.1% fertilization rate). Twenty-nine zygotes
developed to cleaved embryos at the two- to four-cell
stage on the day after fertilization. Embryo transfer was
performed 48 hours after insemination. Clinical pregnancies
were established in three patients (27.2%), as confirmed
by the presence of fetal heartbeat 6 weeks after the
embryo transfer. After 36 weeks of gestation, three fullterm
pregnancies resulted in the delivery of three healthy
infants.
As depicted in Table 1, there was no statistical difference
in terms of patient distribution and clinical outcome between
the patients of the two clinical centers. For statistical
analysis, 2 and Student’s t test were used where appropriate,
and the differences were considered significant when
P.05.
Nonobstructive azoospermia is a frustrating and stressful
dilemma for infertile couples because there are few treatment
options. Treatment with ICSI has alleviated several
forms of severe male infertility, as in patients with cryptozoospermia
or in azoospermic patients, for whom only a
few sperm could be extracted from the testicular biopsies.
The role of FSH in treatment of hypogonadotropic hypogonadal
men has been investigated widely. (3, 4). It has
been reported that administration of FSH combined with
hCG rescues the spermatogenesis and sperm output in
hypogonadotropic hypogonadal azoospermic men (3, 4, 8).
Moreover, treatment with either recombinant (9) or urinary
FSH (10) stimulates spermatogenesis and increases the
spermatogonial population and the number of spermatozoa
in the ejaculate (11) of normogonadotropic patients with
oligozoospermia or severe oligozoospermia. In addition,
administration of FSH to men with azoospermia as a result
of Y-chromosome microdeletion has resulted in the production
of spermatozoa in the ejaculate, which has led to
successful IVF outcomes (12).
Our study demonstrates that gonadotropin treatment of
men who are azoospermic as a result of maturation arrest,
but who have normal hormonal profiles, can improve the
spermatogenesis process in about one fourth (22.4%) of the
treated patients. The mechanism by which some patients
respond to FSH treatment whereas others do not still is
unknown. However, a possible biological explanation may
be that the responding patients may have healthy seminiferous
tubules that can resume spermatogenesis after exogenous
FSH treatment. In addition, it has been shown that men
with normal immunoreactive gonadotropin do not necessarily
have normal biologic gonadotropin activity (13). Therefore, it
could be postulated that at least some of these patients can
benefit from FSH treatment. Interestingly, it has been argued
that a selective decrease in the FSH drive to the seminiferous
tubule leads to a decrease in spermatogenesis, and thus, an
elevation in FSH levels with FSH treatment apparently improves
the spermatogenic process (2).
In this study, before beginning the FSH treatment, we
performed open multiple testicular biopsies on the patients
(6 in each testicle) to achieve an optimal diagnosis and to
locate a site in testis at which spermatogenesis might be
present. It has been claimed that the distribution of spermatogenesis
in the testicles of men with nonobstructive
azoospermia is homogeneous, suggesting that a single diagnostic
biopsy is sufficient to predict the presence of
spermatozoa (14). However, other investigators have demonstrated
that the sperm is produced focally in these cases
and that the chance of finding sperm is correlated to the
testicular pathology and the amount of testicular tissue ex-
TABLE 1
Clinical outcome of patients treated with FSH in two IVF centers.
Characteristic Center 1 Center 2 Total P value
No. of treated men 23 26 49
No. of men responders to FSH treatment (%) 5 (21.7) 6 (24) 11 (22.4) .806
No. of patients who underwent oocyte recovery 5 6 11 .806
No. of retrieved oocytes 29 38 67 .847
No. of injected MII oocytes (%) 24 (82.7) 29 (76.3) 53 (79) .973
No. of fertilized oocytes (%) 15 (62.5) 19 (65.5) 34 (64.1) .256
No. of cleaved embryos (%) 13 (86.6) 16 (84.2) 29 (85.2) .961
No. of embryo transfers 5 6 11 .806
No. of embryos transferred 13 16 29 .749
Mean no. of embryos per patient 2.6 2.7 2.6 .694
No. of clinical pregnancies (%) 1 (20) 2 (33.3) 3 (27.2) .778
% Deliveries 1 2 3 .778
Note: There is no significant difference between the two groups.
Selman. Gonadotropin treatment of azoospermic men. Fertil Steril 2006.
Fertility and Sterility 467