الفهرس 
Intra Cytoplasmic Sperm Injection
Fertilization and Embryo Cleavage after ICSIOnly 14 pages are availabe for public view
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Abstract
nfertility is a common problem, approximately one sixth of marriages are involuntarily childless, while a large number of human pregnancies fail during the implantation stage because the uterine endometrium is unreceptive to the implanting embryo.
Integrins comprise a large family of transmembrane receptors, composed of non-covalently linked α and β subunits. These adhesion molecules are expressed by most cells, including cells involved in immune responses.
In the human endometrium, αvß3 integrin plays an important role in the implantation process & is a well characterized biomarker of uterine receptivity.
The aim of the present study is to find out the utilization of endometrial αvβ3 integrin immunohistochestry scoring as a predictor of embryo implantation in intracytoplasmic sperm injection (ICSI).
This prospective study was performed on fifty (50) patients undergoing intracytoplasmic sperm injection (ICSI) in the Assisted Reproductive Technology Unit of Ain Shams University Maternity Hospital during the period from August 2008 to July 2010.
All patients were 22-36 years old, never conceived before, with no use of any hormonal contraception or IUD, no history of debilitating disease, PID or pelvic operations, with regular menstrual cycles, and they received no hormonal treatment in the month before the study All patients were subjected to the following:
History taking, abdominal and pelvic examination.
1- Trans-vaginal U/S for:
a. Folliculometry (exclusion if abnormal).
b. Exclusion of any ovarian or uterine pathology.
c. Measurement of pre-ovulatory endometrial thickness.
2- Measurement of basal serum FSH, LH, TSH and prolactin, at day 3 of the menstrual cycle.
Each patient had a timed endometrial biopsy during the implantation window, (cycle day 20-24, of a 28 regular cycle; 7 days after ovulation).
All 50 endometrial tissue samples were fixed in 10% formalin solution, embedded in paraffin, sections of 5µm thick were prepared and stained with Haematoxylin-Eosin stain. Then histopathological examination of endometrial biopsies was done at Pathology Department, Ain Shams University Hospital for exclusion of endometrial pathology if any and for proper histological dating according to the histopathological criteria of Noyes et al., (1950).
Paraffin embedded tissue sections of 5µm thick were prepared on positively-charged slides to be stained with anti- αVb3 antibody using Biotin streptavidin immuno-peroxidase technique. Results of integrin staining was scored with a semi-quantitative scoring system by score (0-3) and H score.
In this study, one patient was cancelled (from the 50 selected patients), due to failure of fertilization so they had no embryos transferred back to the uterus.
In this study 17 patients got pregnant (successful implantation), while 32 patients did not get pregnant (failed implantation).
Age ranged from 22 to 36 years with mean age 29.22 years and standard deviation (SD) of 3.51, there was no statistically significant difference (p>0.05) between both (pregnant and non pregnant groups) regarding the age. Also, the age was not correlated to αvβ3 integrin expression among all the patients (p>0.05).
Duration of infertility in patients with failed ICSI was ranging between 1 year and 15 years with mean 6.4 + 3.78 years. There was no statistically significant difference (p>0.05) between both pregnant and non pregnant groups regarding the duration of infertility. Also, the duration of infertility was not correlated to αvβ3 integrin expression among all the subjects (p>0.05).
The maximum number of ova retrieved was 20 and the minimal was 1 ovum with mean 9.69+5.57. There was no statistically significant difference (p>0.05) between both regarding the pregnancy rate. Also, the number of ova retrieved was not correlated to αvβ3 integrin expression among all the patients (p>0.05).
The number of embryos transferred was 1, 2, or 3 embryos, all of grade A, with the mean number of embryos 3 for the pregnant group and 2 for the non pregnant one. Cases with successful implantation showed significantly higher number of embryos transferred when compared to those with failed implantation P<0.001. But there was no correlation between HSCORE, or score (0-3), and number of embryos transferred P>0.05.
All Biopsy endometrial biopsies were taken from day 20 to day 24 from unstimulated cycle, no statistically significant difference (p>0.05) between both regarding the day of Biopsy.
While, there was a positive correlation between HSCORE and date of biopsy Also cases with +ve score in score (0-3) showed significantly higher Date of Biopsy when compared to those with –ve score P<0.05.
Endometrial dating was done and all biopsies showed secretory endometrium from day 19 to day 25, which shows that histological endometrial dating co-incident with menstrual dating, without any evidence of anvolution or luteal phase defect and no endometrial pathology was encountered.
There was no statistically significant difference (p>0.05) between the pregnant and the non pregnant regarding the histological dating.
There was a positive correlation between HSCORE and histological dating P<0.05. And cases with +ve score in score (o-3) showed significantly higher histological dating, when compared to those with –ve score P<0.05.
Immunohistochmical examination of αvß3 integrin of the 49 patients examined revealed positive immunoreactivity in 23(47%) and negative staining in 26 (53&) cases. The αvß3 integrin was localized to the glandular and luminal epithelium in 17 cases. Focal stromal and vessel wall staining was only detected in 2 cases.
Intensity of staining was weak or (+) αvß3 integrin expression was detected in 14 out of the positive 23 cases (60.87%), Moderate or (++) αvß3 integrin expression in 5 cases (21.74%) and strong or (+++) αvß3 integrin expression in 4 cases (17.39%).
In this study, cases with successful implantation showed a significantly higher H score when compared to those with failed implantation P<0.001.
The cut off value of HSCORE was 0.72.
This study showed that the mean intensity of αvβ3 integrin expression, increased progressively with the histological date of the endometrium. H score was reliable to predict endometrial receptivity P<0.0001