The Role of DNA Damage of Granulosa Cell on Oocyte Quality and in Vitro Fertilization Outcome

Suleyman Demirel University60 enrolled

Overview

Deoxyribonucleic acid (DNA) damage of granulosa cells obtained during oocyte retrieval will be evaluated by comet assay in unexplained infertile patients undergoing in vitro fertilization (IVF) treatment. The oocytes will be graded by particular criteria. Fertilization, embryo quality, transfer rate, implantation, clinical pregnancy, pregnancy outcomes (gestational age at delivery, route of delivery, and birthweight etc.) will be recorded as well as demographic data. DNA damage of granulosa cells will be compared between unexplained infertile and control groups. The effect of DNA damage of granulosa cells on fertilization, quality of oocyte and embryo, implantation, and clinical pregnancy will be also evaluated.

Granulosa cells surrounding the oocytes will be mechanically obtained during the oocyte pick-up procedure in women undergoing in vitro fertilization (IVF) treatment due to unexplained infertility. Deoxyribonucleic acid (DNA) damage in these cells will be evaluated by comet assay. The quality of oocytes retrieved during the oocyte pick-up procedure will be graded by particular criteria (zona pellucida thickness, granulation, vacuolization, etc). Fertilization rates, embryo quality by grading, and transfer rates will also be assessed. Implantation and clinical pregnancy rates, and pregnancy outcomes including gestational age at delivery, route of delivery, and birthweight will be recorded as well as demographic data such as age, body-mass index, smoking, alcohol use, employment, coexisting chronic disease, infertility duration, etiology of infertility, treatment protocol, and hormone levels on day 3. Implantation will be evaluated by determination of serum human chorionic gonadotropin (hCG) at day 12 following an embryo transfer. Clinical pregnancy will be diagnosed upon presence of gestational sac on ultrasound examination. DNA damage of granulosa cells will be compared between unexplained infertile group and control group. The effect of DNA damage of granulosa cells on fertilization, quality of oocyte and embryo, implantation, and clinical pregnancy will be also evaluated.

Study Type

OBSERVATIONAL

Enrollment

Conditions

in Vitro Fertilization

Eligibility

Sex: FEMALEMin age: 19 YearsMax age: 44 YearsHealthy volunteers:

Medical Language ↔ Plain English

Inclusion Criteria: * Unexplained infertile patients undergoing in vitro fertilization (IVF) treatment Exclusion Criteria: 1. Chronic systemic disease (rheumatoid arthritis, hypertension, diabetes..) 2. Endocrinopathy (Thyroid, prolactin... abnormalities) 3. Chemotherapy or radiotherapy history 4. Cigarette, alcohol, chronic medication use 5. Diminished ovarian reserve 6. Endometriosis 7. Having any medication use 8. History of any surgical procedure on ovaries and uterus 9. Smoking or alcohol consumption 10. Severe oligoasthenospermia 11. Tubal infertility associated with hydrosalpinx, severe pelvic adhesions, endometriosis or pelvic inflammatory disease

Outcomes

Primary Outcomes

Clinical pregnancy defined as the presence of gestational sac in transvaginal ultrasonographic examination 5 weeks after embryo transfer

Presence of a gestational sac in transvaginal ultrasonographic examination

Time frame: 5 weeks after embryo transfer

Secondary Outcomes

Fertilization defined as the presence of two pronuclei under light microscope one day after intracytoplasmic sperm injection procedure

Presence of two pronuclei under light microscope

Time frame: 1 day after intracytoplasmic sperm injection procedure

Oocyte grade assessed by four parameters (cytoplasmic granulation, properties of the polar body, the perivitelline space and properties of the zona pellucida) under invert microscope 36 hours after human chorionic gonadotropin administration

Oocytes were graded by particular criteria including cytoplasmic granulation, properties of polar body, perivitellin space and zona pellucida. Central cytoplasmic granulation=0, homogenous cytoplasmic granulation=1; large fragmented polar bodies=0, non-fragmented polar bodies with normal size=1; presence of debris in perivitellin space=0, absence of debris in perivitelline space=1; \>15 mm thickness and rough surfaced zona pellucida=0, \<15 mm thickness and smooth surface zona pellucida=1. A total of used 3-4 points were considered as Grade 1 oocytes (best quality), 2-3 points were grade 2 (moderate quality), 0-1 points were considered as grade 3 (poor quality).

Time frame: 36 hours after human chorionic gonadotropin administration

Embryo grade assessed under invert microscope 3 days after intracytoplasmic sperm injection procedure

The quality of embryos was graded from 1 to 3 under inverted microscope 3 days after intracytoplasmic sperm injection procedure. Embryos with even-sized blastomers and/or \<5% fragments were classified as Grade 1 (good quality). Grade 2 embryos (moderate quality) had blastomeres with slightly-moderate size differences and/or 5-50% fragments. Grade 3 embryos (poor quality) had markedly different-sized blastomers and/or \>50% fragments.

Time frame: 3 days after intracytoplasmic sperm injection procedure

Implantation defined as positive serum human chorionic gonadotropin levels 12 days after embryo transfer

Positive human chorionic gonadotropin levels

Time frame: 12 days after embryo transfer