الفهرس 
Review of LiteratureOnly 14 pages are availabe for public view
 |  | from | 181 |  |  |
| | | from | 181 | | |
Abstract
Infertility can be defined as the inability to get pregnancy without any artificial techniques after at least one year of regular unguarded intercourse. Male factor infertility, ovulatory insufficiency and tubal-peritoneal illness are the three main reasons of infertility for the majority of infertile couples. In these cases artificial reproductive methods as intra cytoplasmic sperm injection are recommended.
The most important challenge in ICSI is fertilization failure which may be due to abnormal sperm morphology or absence of chemical mediators which play important role in oocyte activation to complete the process of fertilization.
Artificial oocyte activation may be electrical, thermal and chemical using CA ionophore that has been successfully used to rescue oocyte activation and maintain the intracellular Ca+2 oscillations which is important for fertilization and the embryo development.
The aim of the present work was to evaluate the effect of CA ionophore on the embryonic developmental outcome in female patients with bad quality oocytes as regards the fertilization, cleavage stage (2nd and 3rd day after injection), then blastula stage (day 5 after injection).
The inclusion criteria for the study were released unexplained infertility, male partners with normal semen analysis, female patients who were diagnosed with tubal blockage or hydrosalpinx, polycystic ovarian disorder, endometriosis and premature ovarian failure.
The exclusion criteria for the study were sever male factor, genetic abnormalities and uterine malformations.
Couples were subjected into ICSI trials for the first time, were divided into three groups according to age of the female: group A (20 to 29 years old), group B (30 to 39 years old) and group C (40 to 45 years old).
Female patients were subjected to the recommended ovarian stimulation protocols during their folliculometry, the ovulation and the ovarian retrieval were planned.
Collection of cumulus oocyte complex was done from patients of each group followed by denudation of the COC.
The quality of the oocytes was assessed under the inverted microscope.
The good quality oocytes appeared rounded in shape with regular cell membrane, clear cytoplasm, intact zona pellucida and the polar body was intact in the perivitelline space. While examination of the bad quality oocytes detected oocytes irregular in shape with dark granular cytoplasm, subzonal debris and wide PVS.
To identify the quality of the oocytes some of them were examined in each age group in semithin sections using the light microscope and in ultrathin sections using transmission electron microscope.
The cytoplasm of good quality oocytes contained aggregates of mitochondria and SER with alignment of CG under intact cell membrane. Intact microvilli were seen intersecting the PVS which appeared with normal range.
The cytoplasm of bad quality oocytes included irregular mitochondria, central granules variable in size and multiple vacuoles. Also vacuoles were detected in ZP and PVS was wide in some oocytes while in others was narrow.
According the quality of oocytes, the denuded oocytes from each group were divided into 4 subgroups (I, II, III and IV):
I- Good quality MII oocyte (without CA ionophore treatment)
II- Good quality MII oocyte (with CA ionophore treatment)
III- Bad quality MII oocyte (without CA ionophore treatment)
IV- Bad quality MII oocyte (with CA ionophore treatment)
• In group A, 40 couples were subjected to ICSI cycles; the number of cumulus oocyte complex retrieved was 800 COC. The number of oocytes after denudation was 720 MII oocytes, which were divided after examination into 432 good quality oocytes (AI: 216 oocytes, AII:216 oocytes) and 288 bad quality oocytes (AIII:144 oocytes, A IV:144 oocytes).
• In group B, 40 couples were subjected to ICSI cycles; the number of cumulus oocyte complex retrieved was 480 COC. The number of oocytes after denudation was 432 MII oocytes, which were divided after examination into 260 good quality oocytes (BI:130 oocytes, BII: 130 oocytes) and 172 bad quality oocytes (B III :86 oocytes, B IV: 86 oocytes).
• In group C 38 couples were subjected to ICSI cycles; the number of cumulus oocyte complex retrieved was 115 COC. The number of oocytes after denudation was 92 MII oocytes, which were divided after examination into 48 good quality oocytes (CI: 24 oocytes, C II: 24 oocytes) and 44 bad quality oocytes (C III: 22 oocytes, C IV:22 oocytes).
The denuded oocyte from each group was injected with its corresponding sperm which was examined for normal morphology using the Diff-Quik staining method.
The injected oocytes were cultured in the embryo culture media and the oocytes for sub groups (II, IV) were treated with CA ionophore.
Fertilization was assessed 18 h after ICSI, and normal fertilization was declared when two clearly distinct pronuclei were present.
Embryo quality assessment according to Gardner’s classification was performed at day two, day three and day five after injection using the inverted microscope.
The criteria of good quality embryos in the cleavage stage included that the number of blastomeres must be compatible with embryo age, minor degree of fragmentation less than 20% and absence of the multinucleation.
In the blastula stage the criteria of good quality embryos included full blastocoel cavity, numerous and tightly packed intracellular masses and numerous and cohesive trophectoderm cells.
After collection of data using chia square technique in groups A, B, C, there was increase in the number of fertilized oocytes in the sub groups of bad quality oocytes treated with CA ionophore (IV) with statistically significance correlation only in sub group BIV compared to BIII.
As regard the quality of embryos in the cleavage and blastula stage, there was increase in the number of good quality embryos with statistically significance correlation in sub group AIV compared to AIII, BIV compared to BIII and between CIV compared to CIII especially in day three and blastula stage.
from the present study, it was concluded that using CA ionophore for couples who were subjected to ICSI cycles for the first time especially in females with bad quality oocytes lead to an improvement in the fertilization and the quality of embryos. This meant that there is an increase the possibility and chance of pregnancy.
Recommendations
Post-injection monitoring to obtain pregnancy incidence ratios in terms of pregnancy completion - autumn pregnancy or early childbirth.
Keep the ovarian quality through the following steps:
• Stay away from smoking.
• Mange stress
• Eat healthy
• Achieve normal BMI
• Check any abnormality in the ovarian cycle
• Invest supplements as Coenzyme 10, fish oil and melatonin.
• If any female decided to delay her motherhood, the best way to protect the fertility is oocyte cryopreservation, also it is very important for the young females who receiving chemotherapy.
• Oocyte cryopreservation is a very important process to preserve their fertility.