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Superovulation and intrauterine insemination vs. superovulation alone in the treatment of unexplained infertility. A randomized study.

OBJECTIVE: To assess whether intrauterine insemination (IUI) improves the fertility rates in women with unexplained infertility. STUDY DESIGN: We recruited 68 women with unexplained infertility and allocated them randomly to treatment with three to five cycles of superovulation plus IUI (36 patients) or superovulation alone (32 patients). Superovulation was obtained with clomiphene citrate, human menopausal gonadotropins and human chorionic gonadotropins. RESULTS: The cycle fecundity rate was 10% in patients who underwent superovulation alone and 19% in those treated with superovulation plus IUI (P < .05). CONCLUSION: Our results demonstrate that superovulation plus IUI is more effective than superovulation alone in the treatment of unexplained infertility.

Adult↗

Embryonic characteristics in superovulated mouse strains. Comparative analyses of the incidence of chromosomal aberrations, morphological malformations, and mortality of embryos from two strains of superovulated mice.

Superovulation was induced in an inbred (C3HeB/FeJ) and a random-bred (CD-1) strain of mice and pregnant females were serially sacrificed from 6-15 days of gestation. All embryos were examined chromosomally and by external morphology. A G-banding technique was used to detect specific chromosomal constitutions. The common types of chromosomal aberrations were monosomies and trisomies. Our results showed that the proportion of chromosomally abnormal embryos was significantly greater in the superovulated group of inbred mice, than in the spontaneously ovulating inbred group, an observation not made in the random-bred mice. In each strain, superovulation yielded a significantly higher proportion of morphologically malformed embryos, but no significant difference between strains with respect to this criterion was noted after superovulation. There was a significant increase in the post-implantation embryonic mortality in the random-bred strain but not in the inbred strain following superovulation. This study indicates that each strain has its intrinsic pattern with regard to the fate of embryos following superovulation.

Animals↗

Superovulation in the cow: comparison of oestradiol-17 beta and progesterone patterns in plasma and milk of cows induced to superovulate; relationships with ovarian responses.

Free oestradiol-17 beta, free + conjugated oestradiol-17 beta (total oestradiol-17 beta) and progesterone in milk, and free oestradiol-17 beta and progesterone in plasma were measured in 16 cyclic cows injected with FSH to induce superovulation during the treatment and periovulatory periods. The patterns of steroid secretion were the same in milk as in plasma but at different concentrations. Among oestrogens, the highest concentrations were measured for total oestradiol-17 beta in milk, followed by free oestradiol in plasma and free oestradiol in milk. Progesterone concentrations in milk were higher than in plasma. The peak concentrations of oestrogens were related to ovulation rate: Spearman Rank Correlation coefficient (r.s.) = 0.87 (P less than 0.001), 0.78 (P less than 0.001) and 0.69 (P less than 0.001) for total oestradiol, free oestradiol in milk and free oestradiol in plasma respectively. The increase in progesterone concentrations in milk between the beginning of treatment and prostaglandin injection was negatively correlated with the percentage of viable embryos among those recovered (r.s. = -0.68; P less than 0.001). This was not observed for progesterone in plasma. These results therefore show that the steroid pattern in milk gives a better indication as to the ovarian response to a superovulatory treatment than does the steroid pattern in plasma. In addition the fact that milk samples are easier to obtain and handle than blood plasma have led us to conclude that, to follow the effect of gonadotrophin stimulation, it would be more informative to assay oestradiol-17 beta and progesterone in milk rather than in plasma.

Animals↗

Effects of repeated superovulation and surgical oocyte collection on ovarian response and natural breeding ability of the tammar wallaby (Macropus eugenii).

The aim of this study was to assess the response of a marsupial, the tammar wallaby (Macropus eugenii) to repeated superovulation and surgical oocyte collection and monitor any effects on subsequent natural breeding ability. Animals (n = 5 per group) were superovulated once, twice or three times with pig FSH (pFSH; 6 mg administered twice per day for 4 days) followed by 4 mg pig LH (pLH). There was an interval of either 5-6 weeks (n = 9) or 12 weeks (n = 1) between the first and second superovulation and 13-17 weeks (n = 5) between the second and third superovulation. Oocytes were collected surgically after each treatment. Serum was collected at the time of each treatment to monitor the formation of anti-pFSH and anti-pLH antibodies. Animals were allowed to mate naturally in the season following superovulation treatment(s). There was no significant difference between groups in the number of large follicles (2-5 mm diameter, mean +/- standard error) produced in response to the first (21.2 +/- 4.3), second (18.0 +/- 6.5) or third (29.0 +/- 4.9) superovulation treatment. Eggs were recovered from approximately 80% of follicles that were flushed during laparotomy. There were significant concentrations of anti-pFSH and anti-pLH antibodies (P < 0.05) detected in previously superovulated animals at the time of the second superovulation but not at the time of the third superovulation. The anti-gonadotrophin antibodies present at the time of repeated superovulation did not cause a significant decrease in average number of follicles. All animals produced pouch young in the breeding seasons after repeated superovulation. Combined with other reproductive technologies, repeated superovulation has the potential to increase the production of offspring from rare or valuable marsupials in captivity.

Animals↗

Acceleration of embryo transport in superovulated adult rats.

The etiology of reduced fertility in rodents and humans after exogenous gonadotropin-induced superovulation is unknown. This study examines implantation failure in adult rats induced to superovulate by gonadotropin treatment. After multiple injections of pregnant mare serum gonadotropin (PMSG) followed by human chorionic gonadotropin (hCG), superovulation in adult rats was confirmed on day-1 of pregnancy, however, on day-13, no implantation sites were observed. Daily flushing of oviducts and uteri revealed that the number of embryos recovered from superovulated rats on day-2 was only one-third of that retrieved on day-1. The numbers of embryos retrieved on these days from the oviducts of control rats, however, were almost equivalent. On day-3, no embryos were retrieved from either the oviducts or uteri of superovulated rats, while all embryos from control rats were found in the oviducts. Most embryos recovered from superovulated rats showed normal development. Compared to levels in control rats, serum estradiol (E2) in superovulated rats increased significantly on days-2 and -3 of pregnancy, whereas, serum progesterone (P) levels remained unchanged. Thus, E2/P ratio increased on days-2 and -3 of pregnancy in superovulated adult rats coinciding with the timing of embryo transport acceleration. Our results suggest that superovulated oocytes can be fertilized and reach an early stage of development within the oviduct. Implantation failure in superovulated rats may be due to the accelerated embryo transport resulting from elevated E2/P ratio.

Animals↗

Interrelationships of hormonal and ovarian responses in superovulated heifers pretreated with FSH-P at the beginning of the estrous cycle.

The objective of this study was to determine the relationships between follicle stimulating hormone, (FSH), estradiol (E(2)), and progesterone (P(4)) concentrations in peripheral blood samples and the follicular dynamics prior to and during superovulation in heifers pretreated with FSH-P (10 mg, i.m.) (FSH-P-primed; n=9) or not (saline-primed; n=9) on Day 3 (Day 0 = estrus) of the estrous cycle. On Day 10, all heifers were superovulated with FSH-P (27.7 mg i.m.) in declining dosages over 5 days. Prior to and during superovulation, blood samples were collected one to five times daily, and the follicular dynamics were monitored daily by ultrasonography. Prior to superovulation, profiles of P(4) and E(2) did not differ (P>1) between the saline- and FSH-P-primed heifers. The FSH concentrations in saline-primed heifers decreased from 0.43 +/- 0.05 ng/ml to 0.30 +/- 0.04 ng/ml between Days 3 and 7 and then increased progressively to 0.59 +/- 0.04 ng/ml on Day 10. In contrast (P<0.002), FSH concentrations in the FSH-P-primed heifers remained constant between Days 3 and 10 and averaged 0.41 +/- 0.03 ng/ml. Higher increases in E(2) during superovulation (maximum values, 100 vs 46 pg/ml) and in P(4) after superovulation (maximum values, 39 vs 22 ng/ml) in the saline-than in the FSH-P-primed heifers reflected the greater increase in the number of follicles (>10 mm) and in the number of corpora lutea (CL) in the saline-primed heifers. Prior to the preovulatory luteinizing hormone (LH) peak during superovulation, there was a parallel (P>0.1) decrease in FSH concentrations in the saline- and FSH-P-primed groups. Within heifers partial correlations indicated that E(2) was correlated positively with the number of follicles (>/= 7 mm) and the size of the largest follicle during superovulation (r=0.54 to 0.81; P<0.01). Negative correlations were detected (P<0.01) between FSH and the number of follicles >/=7 mm prior to (r=-0.26) and during superovulation (r=-0.37). The results cofirm earlier reports indicating that priming with FSH-P decreases the superovulatory response in cattle. Interrelationships of hormonal and ovarian responses support the concept that the presence of large dominant follicles prior to superovulation limits the superovulatory response.

Journal Article↗

Pregnancy can be established in superovulated adult rats treated with progesterone and an aromatase inhibitor.

An adult superovulated rat model has been developed and is characterized by high ovulation rates, early morphological degeneration of embryos, complete embryo loss within 48 hours of conception and elevated peripheral estradiol(E2)/progesterone(P4) ratios. In this study, three trials were conducted using the superovulated adult rat model. First, control naturally cycling rats were compared with superovulated rats supplemented with 1 mg P4 on days 0-3 of pregnancy. A sperm positive vaginal smear is designated as day 0 of pregnancy. The P4 treated rats demonstrated improved embryo retrieval on day 1 of pregnancy, continued embryo recovery with a decrease in normal morphologic characteristics of integrity on day 2, with nearly total embryo loss by day 3. On each day, P4 levels were elevated 2-3 times over control. The second trial compared 3 groups of rats, 1) naturally cycling, 2) superovulated unsupplemented and 3) superovulated rats supplemented with 1 mg P4/rat/day and the aromatase inhibitor, 4-hydroxyandrostenedione (4-OHA), 12.5 mg/rat/day. The superovulated unsupplemented rats had no embryo recovery after day 2 of pregnancy, while the P4 and 4-OHA treated rats showed a variable ability to maintain normally developing embryos through day 4 of pregnancy. E2 levels were elevated in both superovulated groups on days 1-4 of pregnancy as were P4 levels on days 2-4. The E2/P4 ratio was significantly lowered only on day 1 of pregnancy in the P4 and 4-OHA treated group. The third trial demonstrated implantation in 50% of the superovulated rats supplemented with P4 and 4-OHA. In conclusion, implantation in the superovulated adult rats can occur with P4 and 4-OHA supplementation, however, this biologic phenomenon could not be explained by obvious changes in peripheral E2 and P4 levels.

Analysis of Variance↗

The role for the uterine insulin-like growth factor I in early embryonic loss after superovulation in the rat.

OBJECTIVES: To examine possible roles of the insulin-like growth factor (IGF) system in increased early embryonic loss after superovulation. DESIGN: Changes in the uterine IGF system were examined in superovulated rats. Insulin-like growth factor I (IGF-I) was infused to the right uterine horns to mimic enhanced IGF-I actions after superovulation. Uterine luminal fluids were collected after IGF-I infusions and embryos were cultured with uterine luminal fluids. MAIN OUTCOME MEASURES: Steroid hormones, IGF-I, IGF binding protein (IGFBP), and IGF-I receptor levels, developmental rate, and cell numbers of embryos. RESULTS: Elevated IGF-I levels and suppressed IGFBP levels were found from days 1 to 3 of pregnancy after superovulation. Uterine luminal fluids of the IGF-I infusion and superovulation groups impaired embryo development in vitro. Anti-IGF-I antibody infusions after superovulation reversed detrimental effects of superovulation. Dialysis of uterine luminal fluids of the IGF-I infusion and superovulation groups before culture improved embryo development. CONCLUSIONS: Enhanced IGF-I actions in the uterus after superovulation may be responsible for the increase of early embryonic loss. The detrimental factor for embryo development seems a small molecule and is likely a local product of the uterus in which IGF-I actions are enhanced.

Animals↗

Efficacy of superovulation and intrauterine insemination in the treatment of infertility. National Cooperative Reproductive Medicine Network.

BACKGROUND: Induction of superovulation with gonadotropins and intrauterine insemination are frequently used to treat infertility. We conducted a large, randomized, controlled clinical trial of these treatments. METHODS: We studied 932 couples in which the woman had no identifiable infertility factor and the man had motile sperm. The couples were randomly assigned to receive intracervical insemination, intrauterine insemination, superovulation and intracervical insemination, or superovulation and intrauterine insemination. Treatment continued for four cycles unless pregnancy was achieved. RESULTS: The 231 couples in the group treated with superovulation and intrauterine insemination had a higher rate of pregnancy (33 percent) than the 234 couples in the intrauterine-insemination group (18 percent), the 234 couples in the group treated with superovulation and intracervical insemination (19 percent), or the 233 couples in the intracervical-insemination group (10 percent). Stratified, discrete-time Cox proportional-hazards analysis showed that the couples in the group treated with superovulation and intrauterine insemination were 3.2 times as likely to become pregnant as those in the intracervical-insemination group (95 percent confidence interval, 2.0 to 5.3) and 1.7 times as likely as those in the intrauterine-insemination group (95 percent confidence interval, 1.2 to 2.6). The couples in the intrauterine-insemination group and in the group treated with superovulation and intracervical insemination were nearly twice as likely to conceive as those in the intracervical-insemination group. CONCLUSIONS: Among infertile couples, treatment with induction of superovulation and intrauterine insemination is three times as likely to result in pregnancy as is intracervical insemination and twice as likely to result in pregnancy as is treatment with either superovulation and intracervical insemination or intrauterine insemination alone.

Abortion, Spontaneous↗

The role of superovulation with menotropins in ovulatory infertility: a review.

The risks of menotropin therapy (ovarian hyperstimulation syndrome, multiple gestation, adnexal torsion) are well known and have been previously described. Superovulation should not be considered for the indications described herein until more traditional therapies for infertility have been tried and found unsuccessful and sufficient time has elapsed for conception to occur. The cost of superovulation is high: the medications are expensive, frequent E2 monitoring and US studies are costly, and pregnancy complications relating to the higher rate of pregnancy loss and multiple gestation may add substantially to the overall cost. Yet, compared with IVF and GIFT, superovulation cycles combined with IUI cost between one third to one sixth that of an IVF cycle. Protocols involving combined CC/hMG/hCG, which reduce the total number of ampules of Pergonal needed per cycle and still provide multiple follicular development, may further reduce costs. There is a growing consensus that superovulation-IUI protocols should be attempted before GIFT and IVF in couples with normal pelvic viscera. There is little doubt that IVF and GIFT cycles are more costly, stressful, and complex. No comparative data have clearly shown IVF and GIFT to be superior to superovulation protocols in ovulatory women with normal pelvic anatomy. In the only study examining this issue published to date, Kaplan et al. retrospectively analyzed all GIFT and superovulation/IUI cycles at a single university center and found GIFT to be three times more efficient. However, the inherent limitations of a nonrandomized, nonprospective study of this kind are obvious as these authors have suggested. Therefore, it may be wise to consider the use of superovulation before assisted reproductive technologies until this issue is settled. It would be interesting to determine if the high PRs reported for couples with unexplained infertility or mild endometriosis in IVF and GIFT cycles in some centers not incorporating superovulation/IUI protocols would hold up if such an approach was routinely followed. Despite the increasing acceptance of superovulation protocols, we must be aware that many of the studies suggesting a role of hMG in treating ovulatory infertile women with normal pelvic anatomy suffer from deficiencies in experimental design. In a payor-driven system, such as in the United States, the difficulties in designing and carrying out scientifically sound clinical studies examining infertility therapies are obvious. The lack of federal or outside funding for the study of infertility issues contributes to the problem. It is our hope that better designed studies examining the role of superovulation in the treatment of ovulatory infertile women with normal pelvic anatomy will be forthcoming.

Female↗

Effects of a GnRH agonist on oocyte number and maturation in mice superovulated with eCG and hCG.

The objective was to investigate the effects of a gonadotropin-releasing hormone agonist (GnRH) on ovulation rate and the number and maturation of oocytes in mice superovulated with equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG). Thirty 3-month-old BALB/C female mice (weight: 25-30 g) were assigned to three experimental groups: control, superovulated, and superovulated with GnRH pretreatment (n=10 per group). Control mice received an i.p. injection of 0.1 ml physiological saline solution. Superovulation was induced with 5 IU eCG (i.p.) and 5 IU hCG 48 h later. Mice in the superovulated with GnRH pretreatment group were given GnRH (20 mg/kg Fertirelin, i.m.), 24 h before superovulation. Thirteen hours after hCG administration, mice were sacrificed by cervical dislocation and blood samples were collected to determine serum progesterone concentration (by radioimmunoassay). Ovaries and oviducts were also harvested to enumerate corpora lutea and cumulus-enclosed oocytes. Progesterone concentrations were not significantly different among groups. The oocyte number and the maturation, ovulation rate, and the number of corpora lutea were higher in GnRH-treated mice than both controls and superovulated mice. In conclusion, GnRH given 24 h before superovulation with eCG-hCG increased the number and maturation of oocytes and the rate of ovulation in mice.

Animals↗

Effect of superovulation prior to mating on milk production performance during lactation in ewes.

Thirty lactating ewes were used to evaluate the effect of superovulation on milk production. Twelve ewes had been injected, prior to mating, with 700 IU of pregnant mare serum gonadotropin; 18 ewes were injected with saline as a control. Thirteen ewes (nine control and four superovulated ewes) were fed at low plane of nutrition; the other ewes (nine control and eight superovulated ewes) were fed at high plane of nutrition. Superovulated ewes, fed at both low and high planes of nutrition, had dramatically higher milk yields (59%), and their milk composition was not changed. Plane of nutrition increased milk lactose and P contents without significant effect on milk production. The increased milk yields in the superovulated ewes were accompanied by increases in dry matter, gross energy intakes, and gross efficiency of milk synthesis. At the end of lactation, superovulated ewes had higher mammary dry fat-free tissue, total DNA, and total RNA. The results demonstrated that superovulation prior to mating dramatically increased milk production and efficiency regardless of plane of nutrition. Increased milk production and efficiency in the superovulated ewes were due to the increased mammary secretory cell numbers and their synthetic activities presumably through the increased endogenous hormonal stimulation of mammary growth and development during pregnancy.

Animal Nutritional Physiological Phenomena↗

Multiple superovulations in N'Dama heifers.

Five N'Dama heifers were superovulated with follicle stimulating hormone (FSH-P or Folltropin) a total of six times each. The superovulations were carried out between ongoing experimental Trypanosoma congolense infections. Twenty-four (80%) of the 30 superovulations had a good ovarian response with 21 (70%) producing an average of 2.7 +/- 0.4 (mean +/- s.e.m.) embryos. The highest embryo production was achieved at the third and fourth superovulation, after which both the number of embryos and their quality declined. The overall pregnancy rate after transfer into Boran (Bos indicus) cow recipients was 50.9%. The uteri of the heifers increased considerably in size throughout the six superovulations which made it difficult to flush some of the animals after the third superovulation. Embryo transfer technology is a useful breeding tool in N'Dama heifers and multiple superovulations can be carried out with success.

Animals↗

After superovulation-intrauterine insemination fails: the prognosis for treatment by gamete intrafallopian transfer/pronuclear stage transfer.

OBJECTIVE: To determine the prognosis for gamete intrafallopian transfer (GIFT)/pronuclear stage transfer (PROST) treatment after prior superovulation-intrauterine insemination (IUI). DESIGN: Matched, retrospective. SETTING: Outpatient university endocrine-infertility program. PATIENTS, PARTICIPANTS: One hundred forty-four women matched for infertility factors and age were studied according to the following three treatment groups: superovulation-IUI only, GIFT/PROST only, or GIFT/PROST after superovulation-IUI. MAIN OUTCOME MEASURES: Per cycle and cumulative pregnancy rates (PRs) were compared utilizing life table analysis. RESULTS: Cumulative PRs (0.408) for superovulation-IUI only were lower than initial (0.469) and cumulative (0.802) cycle fecundity of GIFT/PROST (P = 0.002). Per cycle and cumulative PRs did not differ between GIFT/PROST only versus GIFT/PROST after superovulation-IUI. CONCLUSIONS: Gamete intrafallopian transfer/PROST may be cost-effective when compared with superovulation-IUI. The prognosis for GIFT/PROST success is not negatively affected by earlier superovulation-IUI treatment failure.

Adult↗

Ultrasonic study of follicular dynamics following superovulation in German Merino ewes.

Ewes are commonly superovulated with a single dose of eCG or multiple doses of pFSH. It would be convenient and less expensive to use a single dose of FSH, but results of various trials have been controversial. We wished to investigate ovarian dynamics using ultrasonography after superovulation with a single dose of pFSH and hMG as compared with a single dose of eCG. Estrus was synchronized during the breeding season with fluorogestone acetate-containing intravaginal sponges in adult German Merino ewes (n = 38). They were superovulated with single doses of pFSH (17 mg; n = 10), hMG (600 IU FSH and 600 IU LH; n = 9) or eCG (1250 IU; n = 10) given at the time of sponge removal, or pFSH (17 mg; n = 9) given 36h before sponge removal. Follicular and luteal development were observed by ultrasonic scanning every 8 h from the gonadotrophin injection until the end of estrus, and then once daily until Day 6 after estrus. Jugular venous blood was collected starting immediately before and 1 h after superovulation treatment, then twice daily until the end of estrus and once daily for the following 7 days. Concentrations of estradiol-17beta (E2) and progesterone (P4) were measured in plasma. Differences in the follicular dynamics of the 4 superovulation groups were obvious. The functional duration of the pFSH action was estimated to last approximately 48 h, whereas eCG and hMG were active for up to 72 h. The diameter of the ovulatory follicles proved to be smaller than it was described for unstimulated ewes. Single applications of pFSH or hMG can induce a superovulatory response, although the post-estrus progesterone profile revealed a high premature luteal regression rate in the different superovulation groups. Premature corpus luteum regression could not be seen by ultrasonography at this early stage of the luteal phase, indicating that the technique may fail to detect these corpora lutea in an embryo transfer program. However, ultrasonography represents a suitable method to observe follicular dynamics following different superovulation regimens in sheep.

Administration, Intravaginal↗

Rate of transport and development of preimplantation embryo in the superovulated buffalo (Bubalus bubalis).

This study was designed to ascertain the rate of transport and development of preimplantation embryo in the superovulated buffalo in order to determine the optimum time for their nonsurgical collection. Eighteen Murrah-type buffalo were superovulated with 600 mg NIH-FSH-P1. Luteolysis was induced by administration of PGF2 alpha at 72 (PG + 72) and 84 h (PG + 84) after initiating gonadotrophin treatment and fixed-time AI was done beginning at 36 h post PG + 72 administration and at 12-h intervals thereafter, upto 72 h. Six control buffalo received treatment similar to experimental group except that in place of FSH they received normal saline. For embryo collection, experimental animals were humanely killed at 6-h intervals corresponding to 156 (n = 2), 162 (n = 2), 168 (n = 2), 174 (n = 3), 180 (n = 3), 186 (n = 3) and 192 h(n = 3) after PG + 72 treatment, whereas the control animals were humanely killed at 156 (n = 2), 174 (n = 2) and 192 h (n = 2). Superovulated buffalo had higher number of ovulations than untreated controls (8.78 +/- 5.00 vs 0.67 +/- 0.51) and total ova/embryos recovered was 4.11 +/- 2.46 and 0.67 +/- 0.51, respectively. The high estradiol-17 beta (E2) levels with its prolonged rise may, by leading to reverse peristalsis in the oviduct with a consequent loss of some embryos in the peritoneal cavity, be one of the reasons for our inability to recover nearly 84/158 ova/embryos in the superovulated buffalo. In superovulated animals, nearly all the ova/embryos reached the uterus between 168 and 174 h post PG + 72 treatment or about 134 h (circa 5.5 d) after the onset of superovulatory estrus, suggesting that the ideal time for non-surgical embryo collection in the buffalo is between Days 7 to 8 after PG + 72 treatment or Days 5.5 to 6.0 of the superovulated cycle (estrus = Day 0). Embryo development of superovulated buffalo showed considerable variation as various stages of embryos (8 cell to expanded blastocyst) were recovered from the same donor buffalo, and the rate of development appeared to be 24 to 36 h faster than in cattle.

Animals↗