Fig 1.
Flow chart of the treatment protocol during the study period.
Stage 1: Follicular stimulation (FS) (n = 50). Down-regulation was followed by controlled ovarian stimulation (COS) by gonadotrophin injections (Only Inj Puregon) for fourteen days. The dose was titrated with respect to basal FSH and follicular monitoring by Transvaginal Ultrasound (TVS). Out of the 50 females, 44 females reached the ovarian induction stage (stage 2). Cycle of 4 females was cancelled due to inadequate response to stimulation, whereas two did not opt to continue the treatment. Stage 2: Ovulation induction (OI)(n = 44). On the confirmation of the maturity of at least 3 follicles (measuring18 mm in diameter) by TVS, ovulation was induced by injecting human chorionic gonadotrophin (hCG) 36 ± 1 hour before oocyte pick up (OPU). Maturity of follicles was not confirmed in six females, we could not perform OPU of these females. On the same day, endometrial thickness was measured by a Transvaginal Scan (TVS) in the midsagittal plane by two-dimensional ultrasound with a 7.5 MHz vaginal probe (Hitachi E UB 525; Hitachi, Tokyo Japan) at the thickest endometrial segment [17]. Stage 3: Oocyte Pick Up (OPU) (n = 38). Mature oocytes of females (n = 38) were retrieved 36 hours after hCG injection by vaginal ultrasound probe with 16G adapter and double-lumen oocyte aspiration needle on 14th, 15th, or 16th day of stimulation. All the collected eggs were treated and then transferred to the incubator for about 1–2 hours prior to insemination by ICSI procedures. During this phase, follicular fluid (FF) was acquired from all follicles (measuring 18 mm or more on ultrasound) from each ovary of the patient [18]. Stage 4: Embryo transfer (ET) (n = 27). After confirmation of fertilization and cleavage (18 to 24 hours after egg collection), embryos were graded from 1–5 [19]. The embryo transfer of 27 females (blastocysts) was done on Day 5 using Sims-Wallace Embryo Replacement Catheter, under ultrasound guidance.
Table 1.
Comparison of the descriptive variables between outcome groups.
Fig 2.
Comparison of estradiol (a) and kisspeptin (b) in all stages of stimulation. *Significant with stage 1 at 0.05 ** Significant with stage 1 at 0.01. Stage 1, FS: Follicular stimulation, Stage 2, OI: Ovulation Induction, Stage 3, OPU: Oocyte pickup, Stage 4, ET: Embryo transfer.
Fig 3.
Comparison of estradiol (a) and kisspeptin (b) at various stages of stimulation in all three outcome groups. *Significant with stage 1 at 0.05 ** Significant with stage 1 at 0.01. FS: Follicular stimulation, OI: Ovulation Induction, OPU: Oocyte pickup, ET: Embryo transfer.
Fig 4.
a) Correlation of Follicular estradiol and follicular KP b) Correlation of Follicular KP with number of mature oocytes c) Correlation of Follicular KP with Serum KP3 d) Correlation of Serum KP3 with Serum estradiol e) Correlation of Serum KP3 with endometrial thickness.