   . .   

IVF

30 A 30 B

71.4 B 81.8 A

. .. .. .  ... .... 

..  .         .                 .. .. . 

...  ..  

ﺍﳌﺠﻠﺔ ﺍﻟﺼﺤﻴﺔ ﻟﺸﺮﻕ ﺍﳌﺘﻮﺳﻂ، ﻣﻨﻈﻤﺔ ﺍﻟﺼﺤﺔ ﺍﻟﻌﺎﳌﻴﺔ، ﺍﳌﺠﻠﺪ ﺍﻟﺜﺎﻟﺚ ﻋﺸﺮ، ﺍﻟﻌﺪﺩ ٤، ٢٠٠٧  

 thermore, hyaluronic acid, when added to sperm preparation media, increases sperm From the early beginnings of in vitro fertili- motility and improves retention of sperm zation (IVF) it has been recognized that the motility in long-term incubation of both culture media supplemented with proteins fresh and cryopreserved, thawed human have a direct role in osmoregulation. Pro- spermatozoa [13] teins also serve as a source of energy and Because there have been relatively few as reservoirs for the release of hormones, human studies with hyaluronic acid and be- vitamins and minerals [1]. Because of its cause we have not made use of this medium characteristic high abundance in the female before in our department, we aimed to com- reproductive tract, albumin has traditionally pare the IVF implantation and pregnancy served as the main macromolecule in most rate by using hyaluronic acid and albumin culture media used for in vitro growth of as transfer medium. human embryos [2]. In addition, albumin confers on the culture medium the useful physical properties of lubrication and vis-  cosity, thus promoting ease of handling the embryo and preventing its adherence to the The study design is a prospective double- culture dish [3]. blind study performed from September Because the use of blood-derived al- 2003 to January 2004. As the embryos were bumin can cause both biological variation grown at the embryonic laboratory, the doc- and the possibility of disease transmission, tors and women were unaware which me- several macromolecules, such as polyvinyl- dium had been used. It was conducted at the pyrolidone, polyvinyl alcohol and hyaluron- Research and Clinical Centre for Infertility ic acid have been suggested as alternatives in Yazd, Islamic Republic of Iran and was [4]. Although the potential of polyvinyl- approved by the local ethics committee. pyrolidone and polyvinyl alcohol mac- The study included 60 women undergo- romolecules to promote efficient embryo ing IVF who were divided into Group A development in vitro is still questionable (whose embryos were transferred to a me- [5,6], hyaluronic acid effectively supports dium supplemented with hyaluronic acid) mouse and human embryo development and and a control Group B (whose embryos also their growth [7,8]. were transferred to a medium containing Hyaluronic acid is a naturally exist- albumin). ing macromolecule related to the gly- cosaminoglycans family extra-cellular . matrix and it is a linear polysaccharide [9]. The indication for IVF in groups A and It has been observed that both human [10] B was because of male factors (n = 17, n and bovine [11] embryos possess a surface = 18), tubal factors (n = 9, n = 7) and poly- receptor for hyaluronic acid that can be cystic ovary syndrome (n = 4, n = 5) respec- detected throughout their development up tively. Ovary stimulation was carried out to the blastocyst stage. Research on bovine for all women by a desensitizing protocol embryos suggests that the use of hyaluronic using GnRH-agonist (Suprefact D-65926, acid in the culture medium can increase Hoechst AG, Germany). To suppress pi- the rate of bovine blastocyst and embryo tuitary function, women were treated with development in IVF programmes [12]. Fur- Suprefact 0.5 mg/day given subcutaneously

ﺍﳌﺠﻠﺔ ﺍﻟﺼﺤﻴﺔ ﻟﺸﺮﻕ ﺍﳌﺘﻮﺳﻂ، ﻣﻨﻈﻤﺔ ﺍﻟﺼﺤﺔ ﺍﻟﻌﺎﳌﻴﺔ، ﺍﳌﺠﻠﺪ ﺍﻟﺜﺎﻟﺚ ﻋﺸﺮ، ﺍﻟﻌﺪﺩ ٤، ٢٠٠٧    for 14 days from the mid-luteal phase of the (30 women), embryos were transferred as preceding cycle (day 20–22). As all women routinely done, to a medium containing were ovulatory, they experienced menses albumin 20% (Bayer Corporation, United with Suprefact treatment. After pituitary States) for 10 minutes. All embryo transfers down regulation was confirmed by the ab- were performed using a Labotect catheter sence of any ovarian follicle of > 10 mm (Labotect GmbH, Germany) on day 3 after in size, women underwent gonadotropin oocyte retrieval. stimulation. The luteal phase was supported by pro- Induction of ovulation with human gesterone in oil (Progestan, Nowr pharme- menopausal gonadotropin (HMG) (Menop- Neth, Tehran, Islamic Republic of Iran), 100 ur, Ferring, Germany) was started given in- mg daily administered intramuscularly. tramuscularly in a dosage of 3 ampoules per Patients were tested for serum -hCG day (215 UI of HMG) from day 2 for 7–12 assay 14 days after embryo transfer. If the days depending on the woman’s response. pregnancy test was positive, women were The women were followed by vaginal followed with serial ultrasounds to deter- sonography and monitoring of serum estra- mine fetal viability. Clinical pregnancy was diol (E2) level. When serum E2 level was > defined as the presence of a gestational sac 2000 pmol/L and at least 2 follicles were > on transvaginal ultrasound. Luteal phase 17 mm in diameter, 10 000 units of human support was continued until 10 weeks gesta- chorionic gonadotropin (Organon, Holland) tion. were administered intramuscularly. Trans- vaginal oocyte retrieval was performed . 34–36 hours after hCG administration un- Clinical results of the embryo transfer cy- der transvaginal ultrasound guidance. The cles were compared between the 2 groups. retrieved oocytes were then fertilized (in Data are presented as mean and standard G-fert + albumin 10% for 36–40 hours) deviation (SD). The results were analysed according to sperm quality either by con- using the chi-squared test, Fisher exact test ventional IVF (n = 10) or intracytoplasmic and ANOVA. P-value < 0.05 was consid- sperm injection (n = 20) in each group. ered significant.

.   Sixty (60) women were recruited on the day Table 1 shows some characteristics of the of embryo transfer and signed an informed 2 study groups. Mean age (SD) was 27.5 consent and be included. The inclusion cri- (4.26) years for group A and was 28.60 teria were: age 35 years or younger, having (3.68) years for group B. Mean duration of at least 3 embryos suitable for transfer, and infertility was 7.24 (3.68) years for group having no previous IVF embryo transfer cy- A and 6.93 (3.60) years for group B. Mean cle. The women were randomly divided into embryo transfer was 2.68 (0.66) for group A 2 groups. In group A (30 women), embryos and 2.70 (0.79) for group B (Table 1). were transferred to a medium supplemented In group A, 85 embryos were transferred with 0.5 mL/mL of hyaluronic acid (Em- resulting in 11 pregnancies and in group B, bryoGlue®, Vitrolife, Sweden) for 10 min- 98 embryos were transferred resulting in utes before intrauterine transfer took place. 7 pregnancies. The implantation rate was In group B, that served as a control group

ﺍﳌﺠﻠﺔ ﺍﻟﺼﺤﻴﺔ ﻟﺸﺮﻕ ﺍﳌﺘﻮﺳﻂ، ﻣﻨﻈﻤﺔ ﺍﻟﺼﺤﺔ ﺍﻟﻌﺎﳌﻴﺔ، ﺍﳌﺠﻠﺪ ﺍﻟﺜﺎﻟﺚ ﻋﺸﺮ، ﺍﻟﻌﺪﺩ ٤، ٢٠٠٧  

 supplemented with hyaluronic acid, had  equivalent rates of blastocyst development and equivalent cell numbers compared to     embryos cultured with albumin. In addition, . the implantation and pregnancy rate were     similar in the 2 groups [8].    Our results are similar to another study . conducted in 2003 which suggested that . the clinical pregnancy, implantation and     ongoing pregnancy rates were higher with    hyaluronic acid transfer medium compared . with albumin transfer medium [1]. In an-     other study a significant increase in both    implantation and fetal development rates .  were reported when hyaluronic acid was the only macromolecule in the transfer medium of mouse embryos in comparison to transfer 36.7% in group A and 23.3% in group B, media that were combined with bovine however this difference was not statisti- serum albumin and hyaluronic acid [14]. cally significant (P = 0.26) (Table 2). All The use of hyaluronic acid in the transfer pregnancies were singleton. The abortion medium may offer several advantages in rate was higher in group B (28.6% versus the implantation process, such as a sig- 18.2%), but this was not statistically signifi- nificant increase in both implantation and cant (P 1) (Table 2). fetal development rates. Concern has been expressed about the immediate or late ex- pulsion of embryos after their transfer to the  uterine cavity [15]. However, our experi- ence, as well as that of other researchers This study shows that hyaluronic acid can [1,12,14], suggests that the implantation successfully replace albumin as a human and pregnancy rates were higher in the hy- embryo transfer medium resulting in com- aluronic acid treatment group. We conclude, parable high pregnancy and implantation therefore, that hyaluronic acid medium can rates. successfully replace albumin medium. A study in 2000 showed that the em- bryos, which were developed in media

.        .                            . 

ﺍﳌﺠﻠﺔ ﺍﻟﺼﺤﻴﺔ ﻟﺸﺮﻕ ﺍﳌﺘﻮﺳﻂ، ﻣﻨﻈﻤﺔ ﺍﻟﺼﺤﺔ ﺍﻟﻌﺎﳌﻴﺔ، ﺍﳌﺠﻠﺪ ﺍﻟﺜﺎﻟﺚ ﻋﺸﺮ، ﺍﻟﻌﺪﺩ ٤، ٢٠٠٧   

  . . .      .. .  .               .            .  .       .       .  .     .  .   .          . ..     . ..                       .   .    .         .    . . .   .   .            .     ... . .      ...  .              .          .

ﺍﳌﺠﻠﺔ ﺍﻟﺼﺤﻴﺔ ﻟﺸﺮﻕ ﺍﳌﺘﻮﺳﻂ، ﻣﻨﻈﻤﺔ ﺍﻟﺼﺤﺔ ﺍﻟﻌﺎﳌﻴﺔ، ﺍﳌﺠﻠﺪ ﺍﻟﺜﺎﻟﺚ ﻋﺸﺮ، ﺍﻟﻌﺪﺩ ٤، ٢٠٠٧