الفهرس 
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Abstract
Premature ovarian failure was the occurrence of hyergonadotrophic, hypoestrogenic and amenorrhea before the age of 40 years. Cyclophosphamide is a medication used as chemotherapy and immunosuppresor in certain diseases. Cyclophosphamide treatment caused POF in 42% of treated women. Protection of ovarian function and the repair of injured ovary tissue represented a key concern both before and after chemotherapy treatments. In recent years, many therapeutic agents had been discovered to treat premature ovarian failure induced by chemotherapy. Many investigators mentioned the effect of human amniotic fluid stem cells injection on reversing ovarian damage induced by chemotherapy.
The current study was designed to evaluate the efficacy of human amniotic fluid stem cells injection on premature ovarian damage induced by cyclophosphamide using rat model.
Material and methods
Forty five young adult female albino rats 300-350 grams in weight, between 4-5 weeks of age were used in this study.
The animals were divided into the following groups.
• group Ι (control group):
Fifteen rats were received balanced diet and were divided into 3 groups 5 animals each:
group 1A: the animals received physiological saline by intraperitoneal injection.
group 1B: the animals received physiological saline in the tail vein.
group 1C: the animals were left without treatment.
• group ΙΙ (ovarian damage group)was subdivided into 2 subgroups:
Subgroup ΙΙA: included ten rats that received single intraperitoneal injection of 70 mg/kg cyclophosphamide. The animals were sacrificed one week later (Liu et al., 2012).
Subgroup ΙΙB: included ten rats that received single intraperitoneal injection of 70 mg/kg cyclophosphamide, the animals were sacrificed two weeks later.
• group ΙΙΙ (stem cell treated group):
Consisted of ten rats received single intraperitoneal injection of 70 mg/kg cyclophosphamide, and then 1ml of HAFSCs (approximately 1х10³ cells/µl) was injected via the tail vein 1 week after administration of cyclophosphamide. Then the animals were sacrificed two week later.
At the end of the experimental periods, blood samples were taken from the tail vein then the animals were sacrificed by cervical dislocation under anesthesia by pentobarbital sodium. Their ovaries were dissected out and processed for light microscopic examination using H&E and Masson trichrome stain. Immunohistochemical study for CD105, KI67 and caspase-3 were done .Morphometric and statistical analyses were performed.
Immunohistochemical results of group II revealed positive reaction for caspase-3 and negative immunoreaction of CD105.
In subgroup IIA significant increase in the number of ovarian cyst was detected while the number of growing follicle and corpora lutea showed significant decrease.
In subgroup IIB nearly all ovarian follicles were atretic was absence of corpora lutea .sever inflammatory infiltration of ovarian parenchyma with ovarian fibrosis. These results was associated with significant decrease in estradiol level compared to group I & III
In group III revealed positive CD105 immunoreactivity in OSE, epithelial crypts and interstitium. In some sections the ovary was atrophied and was composed only of interstitial glands. However localized proliferation of OSE with appearance of primordial and growing follicles could be detected in some specimen. Other sections showed more or less more ovarian structure with the presence of recent corpus luteum indicating ovulation. These results were confirmed by significant increase in number of healthy follicles and corpora lutea with significant decrease in the number of atretic follicles and ovarian fibrosis. These results were associated with significant decrease in estradiol level compared to group II.