AOA in Recurrent Fertilization Failure
AOA in Recurrent Fertilization Failure
Background: Despite the high success rate of ICSI, total fertilization failure still occurs
 in 1-3% of all ICSI cycles and can recur in subsequent cycles, even when a sufficient number
 of oocytes and motile spermatozoa are available. Several reports show that the majority of
 couples suffering from ICSI failure benefit from the application of ICSI combined with
 assisted oocyte activation. A variety of artificial activating methods is used in human
 assisted reproduction treatment, including physical, mechanical or chemical stimuli, which
 provoke one or more calcium rises in the oocyte cytoplasm. Study Design: Randomized
 controlled trial. Setting: A university fertility center. Methods: 150 infertile patients who
 underwent ICSI and all had history of recurrent fertilization failure. The patients were
 randomly allocated into 2 equal groups. Group1=75 patients who underwent ICSI without oocyte
 activation. Group2 patients =75 and underwent ICSI Patient underwent ICSI with oocyte
 activation. Reproductive outcomes were compared between both groups. Results: there were
 significant differences between groups regarding number of oocytes retrieved, number of
 mature oocyte, fertilization rate and pregnancy rate. Conclusion: Assisted oocyte activation
 with calcium ionophore results in significant improvement in the fertilization, cleavage and
 pregnancy rates after ICSI.
This study is a randomized controlled trial included 150 ICSI patients with history of
 recurrent fertilization failure. Patients had undergone ICSI trials in ART unit, Al-Azhar
 University, Cairo, Egypt, from July 2017 to November 2018. The study was approved by the
 university medical ethical committee (under registry number 202010454) and all couples had
 signed a written consent before initiation of the study and the treatment cycles. AOA
 candidate couples were counseled regarding the procedure. The patients were selected
 according to the following criteria:

 Inclusion Criteria: -

 1. Age between 20 and 40 years old.

 2. Cases with history of total fertilization failure in previous ICSI cycles

 3. Oocytes with normal morphology.

 4. Male factor infertility. Exclusion Criteria: - Abnormal oocyte morphology degenerated or
 immature oocytes.

 Two groups were randomly designated:

 The first Group: The oocytes were treated by calcium ionophore. This group involved 75 ICSI
 cycles.

 The Second Group: Oocytes were not treated by calcium ionophore. This group involved 75 ICSI
 cycles.

 Methods:

 1. Ovarian stimulation:

 All women received ovarian stimulating drugs according to the ART protocols (long
 agonist, flare up or antagonist protocols). Deca 0.1 was given s.c. daily, for
 down-regulation in short and long protocol while cetrotide 0.25 was given s.c. daily for
 down-regulation in antagonist protocol. Follicular development was monitored by
 ultrasound scanning and serum estradiol. Patients received 10,000 IU of Human Chronic
 Gonadotrophin (HCG) when most of the follicles measured more than 17-20 mm in diameter.

 2. Semen preparation:

 • The husband was asked to submit a semen sample in a sterile plastic container after a
 2 to 3-day period of abstinence and about 2 hours before the ICSI procedure. The
 specimen container must be clean, sterile and wide mouthed to minimize collection error.

 3. Oocyte retrieval:

 -  Under general anesthesia, the oocytes were aspirated by a specialized,
 ultrasound-guided needle (Labotect aspiration catheter, Germany) at 34-36 h after
 HCG injection. Warmed HEPES buffered medium (Irvine Scientific, Irvine, CA, USA)
 was used for handling and washing of oocytes.

 -  After ICSI, the whole injected oocytes were washed with Global total Fertilization
 Medium (Global pharm, Life Global, Brussels, Belgium, Europe) and the patients were
 randomly allocated into two equal groups (75 patients each) by using computer-based
 randomization program. Group one in which, the injected oocytes were transferred to
 a medium that contain 10 µmol of calcium ionophore (GM508 Cult-Active, Gynemed,
 Germany) and were incubated for 10 minutes, then again oocytes were washed with
 Global total Fertilization Medium (Global pharm, Life Global, Brussels, Belgium,
 Europe) and were incubated at 37°C in 6% CO2. In group two, the injected oocytes
 were not submitted to calcium ionophore activation.

 -  18 hours after injection, the whole injected oocytes in both groups were evaluated
 for fertilization, cleavage and quality at the day of embryo transfer (ET).

 Embryo Transfer

 • After ET, luteal phase support was conducted (intramuscularly progesterone injection 100 mg
 daily) for 14 days until pregnancy test.

 After data collection, both groups were compared regarding oocyte number, oocyte
 fertilization rate, number and quality of embryos, implantation rate and clinical pregnancy
 rate.
Inclusion Criteria:

 -  (1) Age between 20 and 40 years old. (2) Cases with history of total fertilization
 failure in previous ICSI cycles (3) Oocytes with normal morphology.

 Exclusion Criteria:

 -  (1) Abnormal oocyte morphology degenerated or immature oocytes. (2) Husbands with
 spermatogenic arrest or Sertoli/Leydig cells only.