Keywords
follicle-stimulating hormone - intracytoplasmic sperm injection implantation - luteinizing
hormone - ovarian stimulation
Palavras-chave
hormônio folículo- estimulante - microinjeção intracitoplasmática de espermatozoides
- hormônio luteinizante - estímulo ovariano
Introduction
For assisted reproductive technology (ART), gonadotropin-releasing hormone (GnRH)
and gonadotropins are routinely administered for controlled ovarian stimulation (COS).
For that, recombinant follicle stimulating hormone (rFSH or follitropin alfa) and
recombinant luteinizing hormone (rLH or lutropin alfa) are the key hormonal stimulus,
which can be used individually or in combination. Follicle stimulating hormone and
LH play distinct but complementary roles in follicle regulation, leading to synergistic
actions in stimulating the recruitment and development of ovarian follicles, increasing
follicle estradiol secretion, and completing oocyte maturation and subsequent ovulation.[1]
Although ovarian stimulation is essential for the success of ART, it is also known
to reduce endogenous FSH and LH releases.[2] Particularly, GnRH antagonists induce a profound pituitary supression, avoiding
premature LH surge. Consequently, recruited follicles are radically deprived of LH
sustenance. Exogenous FSH stimulation will support follicular development in most
patients undergoing ART; however, up to 12% of the patients will not respond to FSH
stimulation alone, which can happen due to the absence of LH.[3] For this subpopulation of patients, there is evidence that LH supplementation to
FSH administration could be advantageous.[3]
[4]
[5]
[6]
[7]
[8]
[9]
[10]
In fact, rLH was originally commercialized to supplement follitropin alfa administration
for specific patients, especially those presenting with severe LH and FSH deficiency,
namely hypogonadotropic hypogonadism. More recently, new products were developed in
a fixed combination of 2:1 (150 IU of rFSH and 75 IU of rLH), under the presupposition
that this is the optimal FSH:LH ratio for the purpose of stimulating follicular development.
Then, it was agreed that patients (i) with previous poor response to ovarian stimulation,
(ii) inadequate ovarian response in the treatment in progress, (iii) aged ≥ 35 years
old could also benefit from LH supplementation.[10]
A recent meta-analysis that included 36 randomized controlled trials investigated
the effectiveness of rLH combined with rFSH for COS compared with rFSH alone in 8,125
women undergoing ART. Moderate quality evidence that the use of rLH combined with
rFSH may lead to more ongoing pregnancies than rFSH alone was observed. No evidence
of a difference between the two regimens was observed in terms of live birth rate.
The authors concluded that the evidence was insufficient to encourage or discourage
stimulation regimens that include rLH combined with rFSH in ART.[11]
To date, there is no evidence that ovarian simulation with rLH improves ART outcomes
in an unselected subpopulation. In addition, there is only one previous study that
investigated cycles in which patients acted as their own controls. The objective of
the present study was to investigate whether patients with a previous rFSH-stimulated
cycle would have improved outcomes with rFSH + rLH stimulation in the following cycle.
Methods
Experimental Design, Patients, and Inclusion and Exclusion Criteria
The present case-control within-subject study included data obtained via chart review
of 228 cycles performed in 114 patients undergoing ICSI between 2015 and 2018 in a
private university-affiliated IVF center. For all patients, rFSH (Gonal-f, Serono,
Geneva, Switzerland) was used for COS in the first ICSI cycle (rFSH group, n = 114), followed by ovarian stimulation with rFSH and rLH (Pergoveris, Merck Serono
S.p.A, Bari, Italy) in the next cycle (rFSH + rLH group, n = 114). Pituitary suppression was achieved with GnRH antagonist (cetrorelix acetate,
Cetrotide; Merck KGaA, Darmstadt, Germany) in both groups.
The inclusion criteria were: couples with primary infertility undergoing their first
rFSH-stimulated ICSI cycle, with intended fresh embryo transfer on day 5 of embryo
development, who underwent a second rFSH + rLH stimulated ICSI cycle, also intending
fresh embryo transfer on day 5 of embryo development.
The exclusion criteria were as follows: Female patients undergoing ICSI cycles with
vitrified/thawed or donated oocytes, surgical sperm retrieval, cryopreserved sperm,
and vitrified/thawed embryo transfer.
Ovarian response to COS and ICSI outcomes were compared between the groups.
All patients signed a written informed consent form. The present study was approved
by the local Institutional Review Board.
Controlled Ovarian Stimulation
For the first ICSI cycle of the patients, COS was started on the 3rd day of the cycle, with the administration of daily doses of r-FSH. For the second
ICSI cycle, on the 3rd day of the cycle, COS was started with the administration of r-FSH + r-LH.
The following steps were the same for both the first and the second ICSI cycles. When
at ≥ 1 follicle ≥ 14 mm was visualized, pituitary blockage was performed using GnRHa.
When ≥ 3 follicles attained a mean diameter ≥ 17 mm and adequate serum estradiol levels
were observed, final follicular maturation was triggered by the administration of
250 µg of r-hCG (Ovidrel, Merck KGaA, Geneva, Switzerland) or GnRH agonist (triptorelin
0.2 mg, Gonapeptyl; Ferring GmbH, Kiel, Germany; or leuprolide acetate 2.0mg, Lupron
Kit, Abbott S.A Societé Française des Laboratoires, Paris, France). Oocyte retrieval
was performed 35 hours later.
Intracytoplasmic Sperm Injection and Embryo Quality and Transfer
Intracytoplasmic sperm injection was performed according to Palermo et al.[12] Embryos were cultured in 50-µL drops culture medium (Global, LifeGlobal, Guilford,
USA) covered with paraffin oil, in a humidified atmosphere under 6% CO2, at 37°C, for 5 days. The embryos were morphologically evaluated on days 3 and 5 of
development. On day 5, 1 to 2 embryos were transferred per patient, depending on maternal
age and embryo quality, using a soft catheter with transabdominal ultrasound guidance.
Clinical Follow-Up
A serum pregnancy test was performed 10 days after embryo transfer. Women with a positive
β human Chorionic Gonadotropin (βhCG) test underwent a transvaginal ultrasound scan
after 2 weeks. Clinical pregnancy was confirmed when at least one intrauterine gestational
sac with fetal heartbeat was detected. Implantation rate was calculated per transferred
embryos. Clinical pregnancy rates were calculated per embryo transfer. Miscarriage
was defined as pregnancy loss before 20 weeks of gestation.
Data Analysis and Statistics
The sample size calculation suggested that 200 cycles would be enough to demonstrate
a 20% effect with 80% power and 5% significance level considering as primary outcome
the implantation rate.
In the first analysis, response to COS, and the outcomes of ICSI were compared between
the rFSH and rFSH + rLH groups (n = 228), using generalized linear models followed by the Bonferroni post hoc test.
Then, data were stratified according to female age (< 35 years old, n = 50, and ≥ 35 years old, n = 178) and response to COS (poor response: ≤ 4 retrieved oocytes, n = 102, and normal response: ≥ 5 retrieved oocytes, n = 126), and were reanalyzed as mentioned above. In all models, female age, body mass
index (BMI) and total FSH dose were included as covariates. No patient has shifted
age categories from the 1st to the 2nd ICSI cycle. Patients that became pregnant in the 1st ICSI cycle and returned for a 2nd cycle desiring another child were not excluded from the analysis, to avoid bias.
Data are expressed as mean ± standard error for continuous variables or as percentages
for dichotomous variables, and p-values. P-value was significant at 5% level (< 0.05).
The analysis was performed using IBM SPSS Statistics for Windows, version 21 (IBM
Corp., Armonk, NY, USA).
Results
All patients completed the follow-up (20 weeks of gestation), and there were no data
missing regarding the reported variables. Higher estradiol levels (1151.73 ± 194.34
pg/mL versus 1909.11 ± 194.34 pg/mL, p = 0.006), oocyte yield (63.41 versus 69.78%, p = 0.045), day-3 high-quality embryos rate (34.13 versus 47.71%, p = 0.029) and implantation rate (18.57 versus 26.47%, p < 0.001), and lower miscarriage rate (33.0 versus 5.0, p = 0.031) were observed in the rFSH + rLH group compared with the rFSH group ([Table 1]).
Table 1
Descriptive analysis of demographics, response to COS and laboratorial ICSI outcomes
of patients in repeated cycles (n = 228)
|
Variables
|
rFSH group (n = 114)
|
rFSH + rLH group (n = 114)
|
p-value
|
|
Female age
|
37.19 ± 0.35
|
37.89 ± 0.35
|
0.160
|
|
Male age
|
39.23 ± 0.63
|
39.96 ± 0.64
|
0.416
|
|
BMI
|
24.88 ± 0.42
|
24.68 ± 0.42
|
0.740
|
|
FSH dose (IU)
|
2826.92 ± 199.67
|
2693.64 ± 198.79
|
0.636
|
|
LH dose (IU)
|
0.0
|
1346.82 ± 34.50
|
NA
|
|
Estradiol level (pg/mL)
|
1151.73 ± 194.34
|
1909.11 ± 194.34
|
0.006
|
|
Cycles triggered with GnRHa
|
9/114 (7.9)
|
10/114 (8.8)
|
0.811
|
|
Follicles (n)
|
9.99 ± 0.70
|
10.38 ± 0.70
|
0.695
|
|
Retrieved oocytes (n)
|
6.37 ± 0.49
|
7.30 ± 0.49
|
0.185
|
|
Oocyte yield (%)
|
63.41 ± 2.24
|
69.78 ± 2.24
|
0.045
|
|
MII oocyte rate (%)
|
67.72 ± 2.53
|
71.48 ± 2.52
|
0.293
|
|
Fertilization rate (%)
|
77.33 ± 2.41
|
73.02 ± 2.37
|
0.202
|
|
Normal cleavage speed rate (%)
|
67.16 ± 3.16
|
73.07 ± 3.11
|
0.182
|
|
D3 high –quality embryos rate (%)
|
34.13 ± 4.37
|
47.71 ± 4.40
|
0.029
|
|
Blastocyst development rate (%)
|
36.72 ± 6.68
|
42.68 ± 5.73
|
0.499
|
|
Frozen embryos (n)
|
2.21 ± 0.61
|
3.05 ± 0.57
|
0.308
|
|
Endometrial thickness (mm)
|
10.32 ± 0.27
|
10.71 ± 0.25
|
0.288
|
|
Embryos transferred (n)
|
2.08 ± 0.09
|
2.04 ± 0.09
|
0.759
|
|
Cycles with embryo transfer (%)
|
70/114 (61.4)
|
69/114 (60.5)
|
0.892
|
|
Implantation rate (%)
|
18.57 ± 0.52
|
26.47 ± 0.62
|
< 0.001
|
|
Pregnancy rate (%)
|
15/70 (21.4)
|
20/69 (29.0)
|
0.303
|
|
Miscarriage rate (%)
|
5/15 (33.0)
|
1/20 (5.0)
|
0.031
|
|
OHSS rate (%)
|
3/114 (2.6)
|
6/114 (5.3)
|
0.308
|
Abbreviations: BMI, body mass index; COS, controlled ovarian stimulation; D3, day
3 of embryo development; ICSI, intracytoplasmic sperm injection; IU, international
unit; MII, metaphase II; NA, not applicable; OHSS, ovarian hyper stimulation syndrome;
rFSH, recombinant follicle stimulating hormone; rLH, recombinant luteinizing hormone.
Note: values are mean ± standard error, unless otherwise noted.
In patients < 35 years old, the implantation rate was significantly higher in the
rFSH + rLH group compared with the rFSH group (21.43 versus 38.46%, p < 0.001) ([Table 2]).
Table 2
Descriptive analysis of demographics, response to COS and laboratorial ICSI outcomes
of patients < 35 years old in repeated cycles (n = 50)
|
Variable
|
rFSH group (n = 25)
|
rFSH + rLH group (n = 25)
|
p-value
|
|
Female age
|
32.00 ± 0.48
|
32.05 ± 0.54
|
0.942
|
|
Male age
|
36.26 ± 1.39
|
36.31 ± 1.66
|
0.981
|
|
BMI
|
25.54 ± 0.98
|
24.58 ± 1.03
|
0.500
|
|
FSH dose (IU)
|
2521.50 ± 138.76
|
2471.05 ± 159.17
|
0.811
|
|
LH dose (IU)
|
0.0
|
1235.53 ± 401.07
|
NA
|
|
Estradiol level (pg/mL)
|
1085.05 ± 399.49
|
1916.20 ± 357.31
|
0.322
|
|
Cycles triggered with GnRHa
|
1/25 (4.0)
|
2/25 (8.0)
|
0.551
|
|
Follicles (n)
|
12.12 ± 1.49
|
13.74 ± 1.71
|
0.475
|
|
Retrieved oocytes (n)
|
8.12 ± 1.09
|
9.84 ± 1.25
|
0.298
|
|
Oocyte yield (%)
|
70.98 ± 4.49
|
75.54 ± 5.16
|
0.505
|
|
MII oocyte rate (%)
|
68.08 ± 4.75
|
67.36 ± 5.44
|
0.920
|
|
Fertilization rate (%)
|
81.08 ± 4.47
|
72.83 ± 5.2
|
0.229
|
|
Normal cleavage speed rate (%)
|
71.79 ± 5.13
|
75.32 ± 5.97
|
0.654
|
|
D3 high-quality embryos rate (%)
|
42.97 ± 9.99
|
40.41 ± 12.24
|
0.871
|
|
Blastocyst development rate (%)
|
41.60 ± 12.12
|
47.44 ± 11.06
|
0.859
|
|
Frozen embryos (n)
|
1.50 ± 0.56
|
2.83 ± 0.64
|
0.118
|
|
Endometrial thickness (mm)
|
10.30 ± 0.59
|
10.87 ± 0.66
|
0.520
|
|
Embryos transferred (n)
|
2.33 ± 0.14
|
2.07 ± 0.17
|
0.223
|
|
Cycles with embryo transfer (%)
|
21/25 (84.0)
|
18/25 (72.00)
|
0.409
|
|
Implantation rate (%)
|
21.43 ± 1.01
|
38.46 ± 1.72
|
< 0.001
|
|
Pregnancy rate (%)
|
6/21 (28.57)
|
9/18 (50.00)
|
0.197
|
|
Miscarriage rate (%)
|
2/6 (33.33)
|
0/9 (0.0)
|
0.083
|
|
OHSS rate (%)
|
0/25 (0.0)
|
1/25 (4.0)
|
0.999
|
Abbreviations: BMI, body mass index; COS, controlled ovarian stimulation; D3, day
3 of embryo development; ICSI, intracytoplasmic sperm injection; IU, international
unit; MII, metaphase II; NA, not applicable; OHSS, ovarian hyper stimulation syndrome;
rFSH, recombinant follicle stimulating hormone; rLH, recombinant luteinizing hormone.
Note: values are mean ± standard error, unless otherwise noted.
In patients aged ≥ 35 years old, higher estradiol levels (1161.80 ± 215.94 pg/mL versus
1966.55 ± 220.13 pg/mL, p = 0.009), oocyte yield (61.28%versus 68.62%, p = 0.038), day-3 high-quality embryos rate (32.01 versus 48.81%, p = 0.013), and implantation rate (17.35 versus 23.64%, p < 0.001) were observed in the rFSH + rLH group compared with the rFSH group ([Table 3]).
Table 3
Descriptive analysis of demographics, response to COS and laboratorial ICSI outcomes
of patients aged ≥ 35 years old in repeated cycles in patients (n = 178)
|
Variable
|
rFSH group (n = 89)
|
rFSH + rLH group (n = 89)
|
p-value
|
|
Female age
|
38.65 ± 0.29
|
39.06 ± 0.28
|
0.303
|
|
Male age
|
40.10 ± 0.68
|
40.67 ± 0.66
|
0.549
|
|
BMI
|
24.70 ± 0.47
|
24.70 ± 0.45
|
0.995
|
|
FSH dose (IU)
|
2913.69 ± 248.86
|
2738.16 ± 2369.51
|
0.611
|
|
LH dose (IU)
|
0.0
|
1369.08 ± 359.66
|
NA
|
|
Estradiol level (pg/mL)
|
1161.80 ± 215.94
|
1966.55 ± 220.13
|
0.009
|
|
Cycles triggered with GnRHa
|
8/89 (9.0)
|
8/89 (9.0)
|
> 0.999
|
|
Follicles (n)
|
9.39 ± 0.77
|
9.71 ± 0.75
|
0.772
|
|
Retrieved oocytes (n)
|
5.88 ± 0.54
|
6.79 ± 0.53
|
0.227
|
|
Oocyte yield (%)
|
61.28 ± 2.54
|
68.62 ± 2.46
|
0.038
|
|
MII oocyte rate (%)
|
67.61 ± 2.95
|
72.33 ± 2.83
|
0.248
|
|
Fertilization rate (%)
|
76.20 ± 2.81
|
73.06 ± 2.65
|
0.417
|
|
Normal cleavage speed rate (%)
|
65.72 ± 3.78
|
72.65 ± 3.57
|
0.184
|
|
D3 high-quality embryos rate (%)
|
32.01 ± 4.83
|
48.81 ± 4.69
|
0.013
|
|
Blastocyst development rate (%)
|
39.06 ± 7.34
|
45.10 ± 6.24
|
0.531
|
|
Frozen embryos (n)
|
2.42 ± 0.75
|
3.09 ± 0.67
|
0.508
|
|
Endometrial thickness (mm)
|
10.33 ± 0.30
|
10.68 ± 0.26
|
0.386
|
|
Embryos transferred (n)
|
1.98 ± 0.11
|
2.04 ± 0.10
|
0.712
|
|
Cycles with embryo transfer (%)
|
49/89 (55.06)
|
51/89 (57.30)
|
0.698
|
|
Implantation rate (%)
|
17.35 ± 0.60
|
23.64 ± 0.66
|
< 0.001
|
|
Pregnancy rate (%)
|
9/49 (18.37)
|
12/51 (23.53)
|
0.508
|
|
Miscarriage rate (%)
|
3/9 (33.33)
|
1/12 (8.33)
|
0.140
|
|
OHSS rate (%)
|
3/89 (3.4)
|
5/89 (5.6)
|
0.469
|
Abbreviations: BMI, body mass index; COS, controlled ovarian stimulation; D3, day
3 of embryo development; ICSI, intracytoplasmic sperm injection; IU, international
unit; MII, metaphase II; NA, not applicable; OHSS, ovarian hyper stimulation syndrome;
rFSH, recombinant follicle stimulating hormone; rLH, recombinant luteinizing hormone.
Note: values are mean ± standard error, unless otherwise noted.
In patients with poor response to COS (≤ 4 retrieved oocytes), oocyte yield (56.82
versus 63.29%, p = 0.001), mature oocytes rate (69.87 versus 78 + 12%, p < 0.001), normal cleavage speed (62.5 versus 75.83%, p < 0.001), implantation rate (10.00 versus 20.45%, p < 0.001) and miscarriage rate (100 versus 0.00%, p < 0.001) were improved in the rFSH + rLH group compared with the rFSH group ([Table 4]).
Table 4
Descriptive analysis of demographics, response to COS and laboratorial ICSI outcomes
in repeated cycles in patients with poor response to COS (≤ 4 retrieved oocytes) (n = 102)
|
Variable
|
rFSH group (n = 51)
|
rFSH + rLH group (n = 51)
|
p-value
|
|
Female age
|
38.37 ± 0.54
|
38.93 ± 0.58
|
0.481
|
|
Male age
|
39.90 ± 0.89
|
39.97 ± 0.98
|
0.959
|
|
BMI
|
25.11 ± 0.45
|
24.23 ± 0.50
|
0.194
|
|
FSH dose (IU)
|
3051.60 ± 456.34
|
2536.05 ± 492.09
|
0.442
|
|
LH dose (IU)
|
0.0
|
1268.02 ± 442.97
|
NA
|
|
Estradiol level (pg/mL)
|
596.24 ± 101.87
|
725.51 ± 111.24
|
0.391
|
|
Cycles triggered with GnRHa
|
0/51 (0.0)
|
0/51 (0.0)
|
> 0.999
|
|
Follicles (n)
|
4.65 ± 0.30
|
4.33 ± 0.33
|
0.472
|
|
Retrieved oocytes (n)
|
2.29 ± 0.16
|
2.47 ± 0.18
|
0.481
|
|
Oocyte yield (%)
|
56.82 ± 1.31
|
63.29 ± 1.34
|
0.001
|
|
MII oocyte rate (%)
|
69.87 ± 1.34
|
78 + 12 ± 1.56
|
< 0.001
|
|
Fertilization rate (%)
|
79.46 ± 1.68
|
81.0 ± 1.8
|
0.533
|
|
Normal cleavage speed rate (%)
|
62.5 ± 1.08
|
75.83
|
< 0.001
|
|
D3 high-quality embryos rate (%)
|
32.47 ± 6.44
|
49.14 ± 7.32
|
0.087
|
|
Blastocyst development rate (%)
|
32.81 ± 7.23
|
33.20 ± 6.41
|
0.967
|
|
Frozen embryos (n)
|
0.71 ± 0.17
|
0.69 ± 0.19
|
0.957
|
|
Endometrial thickness (mm)
|
10.22 ± 0.45
|
10.10 ± 0.50
|
0.861
|
|
Embryos transferred (n)
|
1.68 ± 0.12
|
1.68 ± 0.13
|
0.992
|
|
Cycles with embryo transfer (%)
|
25/51 (49.02)
|
26/51 (50.98)
|
0.836
|
|
Implantation rate (%)
|
10.00 ± 0.63
|
20.45 ± 0.96
|
< 0.001
|
|
Pregnancy rate (%)
|
3/25 (12.00)
|
6/26 (23.08)
|
0.332
|
|
Miscarriage rate (%)
|
3/3 (100)
|
0/6 (0.0)
|
< 0.001
|
|
OHSS rate (%)
|
0/51 (0.0)
|
0/51 (0.0)
|
0.999
|
Abbreviations: BMI, body mass index; COS, controlled ovarian stimulation; D3, day
3 of embryo development; ICSI, intracytoplasmic sperm injection; IU, international
unit; MII, metaphase II; NA, not applicable; OHSS, ovarian hyper stimulation syndrome;
rFSH, recombinant follicle stimulating hormone; rLH, recombinant luteinizing hormone.
Note: values are mean ± standard error, unless otherwise noted.
In patients with normal response to COS (≥ 5 retrieved oocytes), higher estradiol
levels (1725.74 ± 303.65 pg/mL versus 2788.37 ± 281.12 pg/mL, p = 0.010), oocyte yield (75.37 versus 82.69%, p = 0.006), and implantation rate (23.33 versus 29.35%, p < 0.001) were observed in the rFSH + rLH group compared with the rFSH group ([Table 5]).
Table 5
Descriptive analysis of demographics, response to COS and laboratorial ICSI outcomes
in repeated cycles in patients with normal response to COS (≥ 5 retrieved oocytes)
(n = 126)
|
Variable
|
rFSH group (n = 63)
|
rFSH + rLH group (n = 63)
|
p-value
|
|
Female age
|
36.24 ± 0.44
|
37.27 ± 0.42
|
0.092
|
|
Male age
|
38.78 ± 0.86
|
39.95 ± 0.83
|
0.322
|
|
BMI
|
24.68 ± 0.66
|
24.94 ± 0.60
|
0.770
|
|
FSH dose (IU)
|
2648.61 ± 75.42
|
2789.08 ± 71.04
|
0.175
|
|
LH dose (IU)
|
0.0
|
1394.54 ± 308.62
|
NA
|
|
Estradiol level (pg/mL)
|
1725.74 ± 303.65
|
2788.37 ± 281.12
|
0.010
|
|
Cycles triggered with GnRHa
|
9/63 (14.3)
|
10/63 (15.9)
|
0.803
|
|
Follicles (n)
|
14.32 ± 0.91
|
14.04 ± 0.86
|
0.826
|
|
Retrieved oocytes (n)
|
9.67 ± 0.61
|
10.23 ± 0.57
|
0.503
|
|
Oocyte yield (%)
|
75.37 ± 1.99
|
82.69 ± 1.78
|
0.006
|
|
MII oocyte rate (%)
|
73.33 ± 1.43
|
71.8 ± 1.69
|
0.489
|
|
Fertilization rate (%)
|
78.31 ± 2.65
|
72.40 ± 2.51
|
0.105
|
|
Normal cleavage speed rate (%)
|
70.02 ± 3.16
|
72.54 ± 3.00
|
0.565
|
|
D3 high-quality embryos rate (%)
|
35.80 ± 5.96
|
46.78 ± 5.45
|
0.174
|
|
Blastocyst development rate (%)
|
41.79 ± 11.63
|
53.29 ± 9.59
|
0.445
|
|
Frozen embryos (n)
|
3.71 ± 0.99
|
4.23 ± 0.80
|
0.680
|
|
Endometrial thickness (mm)
|
10.40 ± 0.33
|
10.98 ± 0.27
|
0.182
|
|
Embryos transferred (n)
|
2.29 ± 0.107
|
2.21 ± 108
|
0.603
|
|
Cycles with embryo transfer (%)
|
45/63 (71.43)
|
42/63 (66.67)
|
0.513
|
|
Implantation rate (%)
|
23.33 ± 0.72
|
29.35 ± 0.80
|
< 0.001
|
|
Pregnancy rate (%)
|
12/43 (27.91)
|
13/42 (30.95)
|
0.579
|
|
Miscarriage rate (%)
|
2/12 (16.67)
|
1/13 (7.69)
|
0.425
|
|
OHSS rate (%)
|
3/63 (4.76)
|
6/63 (9.52)
|
0.299
|
Abbreviations: BMI, body mass index; COS, controlled ovarian stimulation; D3, day
3 of embryo development; ICSI, intracytoplasmic sperm injection; IU, international
unit; MII, metaphase II; NA, not applicable; OHSS, ovarian hyper stimulation syndrome;
rFSH, recombinant follicle stimulating hormone; rLH, recombinant luteinizing hormone.
Note: values are mean ± standard error, unless otherwise noted.
Discussion
In the present study, we observed that COS with rFSH + rLH resulted in higher estradiol
levels, oocyte yield, day-3 high-quality embryos rate and implantation rate, and lower
miscarriage rate compared with COS with rFSH only. The only previous study that has
investigated the effect of adding rLH to stimulation in patients with a previous cycle
stimulated with rFSH alone showed lower fertilization rates associated with rLH supplementation.[13]
The use of rLH during COS is a matter of debate in the literature that has produced
controversial results. In studies that investigated the benefits of adding LH to FSH
stimulus in women with normal response to COS, higher levels of estradiol[14]
[15]
[16]
[17]
[18] and progesterone,[15] higher rate of high-quality embryos,[15] a smaller number of cycles cancelled,[14] increased pregnancy rate,[14] and less incidence of OHSS[14] were observed compared with stimulus with rFSH alone. One study demonstrated a negative
impact of LH supplementation on oocyte maturation and fertilization.[13] Conversely, several studies reported no difference in the outcomes of cycles when
rFSH alone was compared with rFSH + rLH.[8]
[19]
[20]
[21]
[22]
In patients with poor response to COS, stimulation with rFSH + rLH resulted in higher
pregnancy, implantation, and live birth rates when compared with stimulation with
rFSH alone or human menopausal gonadotropin.[23] Another study showed that stimulation with rFSH + rLH yielded higher rate of high-quality
embryos.[19] These results suggest that poor response to COS could be related to LH insufficiency,
and rLH supplementation might rescue oocyte competence that, in turn, could lead to
the development of viable embryos, thus increasing pregnancy outcomes. On the other
hand, some studies reported no significant differences in ICSI outcomes when comparing
the two stimulation regimens in poor responder patients.[24]
[25]
Significantly increased implantation ratse[6]
[8]
[9] and live birth rates[9] have been observed in older women stimulated with rFSH + rLH when compared with
their nonsupplemented counterparts.[8] Moreover, treatment with rLH significantly reduced total FSH consumption,[8] confirming that FSH and LH act synergistically. Conversely, Fábregues et al.[26] showed that rLH supplementation did not increase ovarian response to COS and implantation
rates in patients of older reproductive ages. Marrs et al.[21] observed similar pregnancy rates in young and older women receiving rFSH + rLH;
however, pregnancy rates in women ≥ 35 years old receiving rFSH alone significantly
declined when compared with those of women < 35 years old, suggesting that these patients
might benefit from the addition of rLH. It has been suggested that younger women possess
a higher number of LH receptors compared with older women and, therefore, do not require
LH supplementation, while LH supplementation in older women secures a sufficient LH-induced
response.[8] Moreover, ovarian androgen secretion is also diminished in older women, suggesting
an age-related decline in ovarian response to stimulation with LH.[8]
Few studies investigated the potential benefits of adding rLH to rFSH in patients
with reduced serum LH concentrations. Lisi et al.[27] showed an increased implantation rate in women with LH concentration < 1.0 IU/l
at downregulation who received rLH supplementation, suggesting that, in the experience
of profound LH downregulation, rLH supplementation might be beneficial. In opposition,
Humaidan et al.[8] observed increased implantation rates when LH supplementation was used in patients
with endogenous LH concentrations ≥ 1.99 IU/l. One study failed to demonstrate association
between rLH supplementation and improved outcomes.[28]
The lack of consensus in the aforementioned literature has led to the publication
of several meta-analyses, which also came to conflicting conclusions. The meta-analyses
suggested that rFSH+rLH results in shorter stimulation length and fewer rFSH consumption,[29] and yields higher estradiol levels[29]
[30] and higher number of mature oocytes.[30] While several meta-analysis showed no significant differences in implantation,[2]
[29]
[30] pregnancy,[2]
[29]
[30] and live-birth rates,[2]
[11]
[30]
[31] others have demonstrated higher implantation rates,[3] pregnancy rates,[3]
[32] ongoing pregnancy rates,[11] and lower miscarriage rates[11] in the recombinant LH-supplemented regimen.
For poor responder patients, an increase in clinical pregnancy rate was observed in
favor of supplementing rLH.[2]
[32] In addition, poor responders showed significantly more retrieved oocytes with rFSH + rLH
compared with rFSH alone.[32]
The disparity found in the literature may be due to (i) LH administration start (beginning
of treatment or late phase), (ii) type of GnRH analogue used (agonist or antagonist),
(iii) starting dose of gonadotropin and gonadotropin dose adjustment during COS, (iv)
heterogeneous definition of poor ovarian response, and (v) heterogeneous cutoff values
for advanced maternal age (35 or 36 years old).
The possible mechanisms behind the benefits offered by rLH supplementation are improved
oocyte competence and endometrial receptivity. Lower levels of cumulous cell apoptosis
have been demonstrated in cycles with rLH supplementation as compared with cycles
with rFSH alone,[33] which can reflect enhanced oocyte competence. In addition, LH stimulates CYP17 to
convert progesterone into androgens, which in turn can be aromatized to estrogens.
The supplementation with LH decreases the chance of a premature progesterone rise
prior to luteinization, thus benefitting the endometrium and increasing the chance
of implantation and clinical pregnancy.[34] Finally, the addition of rLH may improve follicular insulin sensitivity, leading
to decreased androgen levels through a cascade mediated by increased production of
adiponectin. This favorable setting may culminate in enhanced follicular maturation,
ovulation, and fertilization capacity.[35]
This is a retrospective study with its inherent limitations and bias. In addition,
although the sample size was adequate for the analysis of the general group, the present
study is underpowered for subgroups analyses. The present study was limited by its
small sample size but creates a rationale to conduct randomized studies with larger
casuistic to draw concrete conclusions about the use of rFSH and rLH for ovarian stimulation
in patients with cycles stimulated with rFSH alone. The results presented here might
provide another tool for the clinician to use in the decision-making process regarding
the trigger regimen. The most important take home message is that the outcomes of
ICSI cycles from unselected patients can be improved in a following cycle with the
use of LH supplementation for ovarian stimulation.
Conclusion
In conclusion, ovarian stimulation with LH supplementation results in higher implantation
rates, regardless of maternal age and response to COS, compared with cycles stimulated
with rFSH only. Improvements were also observed for ICSI laboratory outcomes and miscarriage
rate when the patients were stratified by age and number of retrieved oocytes. Despite
being encouraging, due to the retrospective nature of the present study, these results
should be confirmed in randomized controlled trials.
