The Role of Ovarian PRP Therapy in Poseidon Poor Responders Women Undergoing ICSI Cycle

1. Materials And Methods 2.1. Study design This study was conducted at a private fertility center in Karbala, Iraq, from Jan 2024 to Feb 2025. All participants provided written informed consent prior to enrollment, and ethical approval was obtained from the College of Medicine, Karbala University (reference number 25-8).

2.2. Participants A total of 102 Poseidon POR women were enrolled where Inclusion criteria were infertile Poseidon groups 2, 3, 4 women with no history of chronic illness or pelvic pathology. All male partners were either normozoospermic or had mild OAT delivered fresh ejaculated samples for ICSI.

Exclusion criteria were ovarian failure due to sex chromosome abnormalities, current use of anticoagulants and history of bleeding disorders. Participants were assigned into two groups: Study group (n=50), who underwent ovarian PRP injections once or twice times within six months prior to ICSI cycle, and control group (n=52), who did not receive PRP therapy.

2.3. PRP preparation and injection Twenty mL of peripheral blood was drawing under sterile conditions using Lora PRP tubes (Modern Technology Company). Frist the blood was centrifuged at 1600 rpm for 10 minutes (soft spin). Then the plasma and superficial buffy coat were transferred into a sterile tube containing no anticoagulant and centrifuged again at 3500 rpm for 5 minutes (hard spin). The upper 2/3rd (platelet-poor plasma, PPP) were discarded, and the platelet pellets were homogenized in the lower 1/3rd of plasma to create 3 mL of platelets concentrate of 3-5 times higher than basal and injected within one hour of preparation (11). During early-mid follicular phase, using TV-US guidance and conscious sedation, 1.5 ml of PRP was injected into each ovary's subcortical layer using a 17-gauge ovum pick-up needle. A second PRP injection was administered when subsequent ORTs (AMH,AFC) and hormone levels (FSH, LH, E2) showed suboptimal results in patient's next menstrual cycles.

2.4. ICSI cycle outcomes All patients starting same COS protocol began on CD3 using (300-450 iu) of Gonal-F (r-FSH); Merck ± (u-HMG);LG in flexible antagonist protocol, (Cetrotide); Merck. Final oocyte maturation was triggered by 250 mcg r-hCG (Ovitrelle; Serono) ± 0.2 mg GnRH agonist (Decapeptyl; Merck) 35 hours before oocyte retrieval when dominant follicles reached (17-18) mm. Embryological outcomes including; MII oocytes, embryos quantity and quality were compared in both group. In all embryo transfer cycles, 2-4 embryos grade A± B of age 2-5 day were transferred. The clinical outcomes included pregnancy rate (serum β-HCG ≥100 mIU/mL), miscarriage rate (the loss of a pregnancy before 24 weeks), Live Birth rates (delivery of a living baby after 24 weeks) and overall cycle outcomes were compared in both groups.