Standardization and Application of Pure Platelet-Rich Plasma in Improving Sperm Function

Standardization and Application of Pure Platelet-Rich Plasma in Improving Sperm Function

Phase 1: Standardize and validate pure platelet-rich plasma (P-PRP) protocol

Aims:

To validate a standardized double-spin protocol for preparing pure platelet-rich plasma (P-PRP) in an IVF laboratory setting by assessing its platelet yield, purity, and reproducibility.

Outcomes

Primary outcome Platelet concentration in PRP Absolute platelet count in P-PRP and fold-increase compared to whole blood Secondary outcome Platelet recovery (%): (Platelet count in P-PRP × P-PRP volume) / (Whole blood platelet count × blood volume) × 100 Purity: White blood cell (WBC) concentration in P-PRP Growth factors :e.g.. VEGF, PDGF-AB, TGF-β, IGF-1… concentrations in plasma of PPP and P-PRP preparation Reproducibility Metrics: Coefficient of variation (CV%), intraclass correlation coefficient (ICC), and Bland-Altman limits of agreement for technical and biological duplicates.

Phase 2: Examine the effect of pure platelet-rich plasma (P-PRP) on sperm parameters Hypotheses and objectives Hypotheses The investigator hypothesizes that co-culture of semen with pure platelet-rich plasma (P-PRP) enhances sperm quality by increasing motility compared to standard culture medium without P-PRP (control group).

Primary outcome

- To compare total sperm motility after 24 hours of co-culture between semen samples co-cultured with P-PRP versus a control group (without P-PRP).

Secondary outcome

• To compare sperm parameters after other timepoints of co-culture between semen samples co-cultured with P-PRP versus a control group (without P-PRP).
• To compare the morphology and DNA Fragmentation Index (DFI) in semen samples after 24-hour co-culture with P-PRP versus control.

Inclusion Criteria:

• Female volunteer aged 18-45 years, willing to donate their blood
• Adequate sperm samples (raw sample TMS > 5 x 106)
• BMI <30 kg/m2

Exclusion Criteria:

• Concurrent administration of other agents such as prednisolone, intravenous immunoglobulin, or G-CSF
• Drug addiction
• Underlying uncontrolled genital infections, diabetes or hypertension, chromosomal or uterine abnormalities, genetic, hematologic, immunological, or endocrine disorders
• Chronic disease, systemic disease, or cancers
• Blood diseases (sepsis, thrombocytopenia)
• Do not agree to donate blood
• Use of supplements containing antioxidants within the past 3 months
• Administration of anticoagulants or NSAIDs at least 7 days before P-PRP infuse
• Haemoglobin < 11g/dL, platelet count < 150000 mm3