Platelet-Rich Plasma (PRP) — Step-by-Step Preparation Guide

What is PRP?

• Platelet-Derived Growth Factor (PDGF)
• Epidermal Growth Factor (EGF)
• Transforming Growth Factor (TGF)
• Vascular Endothelial Growth Factor (VEGF)

Equipment & Supplies

Step 1 — Patient Preparation

• Instruct the patient to avoid NSAIDs, aspirin, and platelet-inhibiting medications for at least 7–10 days before the procedure.
• Ensure adequate hydration on the day of collection.
• Obtain informed consent.

Step 2 — Blood Collection

1. Identify a suitable antecubital vein.
2. Draw 10–40 mL of venous blood (volume depends on desired PRP yield — 10 mL is standard for scalp/facial treatment).
3. Collect blood directly into tubes pre-loaded with anticoagulant:
   • ACD-A (Acid Citrate Dextrose - Solution A) is the anticoagulant of choice — it best maintains platelet viability and prevents premature activation.
   • Alternatively, citrate phosphate dextrose-adenine (CPDA) can be used (approximately 10% less effective).
4. Gently invert the tube 8–10 times to mix with anticoagulant. Do not shake — this can activate platelets prematurely.

Step 3 — First Centrifugation (Soft Spin)

┌─────────────────────────────┐
│  Upper layer (PPP)          │  ← Platelet-poor plasma (pale yellow)
│─────────────────────────────│
│  Buffy coat (thin layer)    │  ← WBCs + platelets (white line)
│─────────────────────────────│
│  Bottom layer               │  ← Red blood cells (dark red)
└─────────────────────────────┘

• Leucocyte-poor PRP (P-PRP): Collect only the upper layer + superficial buffy coat
• Leucocyte-rich PRP (L-PRP): Include the full buffy coat layer

Step 4 — Transfer of Supernatant

1. Using a syringe and 3-way cannula, carefully aspirate the upper plasma layer and buffy coat into a fresh tube.
2. Avoid disturbing the RBC layer — contamination degrades PRP quality.

Step 5 — Second Centrifugation (Hard Spin)

• Top 2/3 = Platelet-poor plasma (PPP) — discard this
• Bottom 1/3 = Platelet-rich plasma (PRP) concentrate + cell pellet

Step 6 — Resuspension / Final PRP Collection

1. Carefully remove and discard the top two-thirds of the supernatant (PPP).
2. Gently resuspend the platelet pellet in the remaining small volume of plasma by pipetting up and down.
3. This yields your final PRP concentrate (typically 2–5 mL from a 10–30 mL draw).

Step 7 — Activation (Optional, Indication-Dependent)

Once activated, fibrinogen converts to fibrin and a fibrin clot begins to form. Activated PRP must be injected within minutes before it solidifies.

Step 8 — Quality Check (Optional but Recommended)

• Final PRP should appear as a pale yellow/straw-colored solution (slightly more turbid than plasma).
• Platelet count in the final PRP should ideally be ≥1,000,000 platelets/µL (4–7× baseline).

Step 9 — Injection

• Inject intradermally or subdermally into the scalp using a 30G needle, ~0.1 mL per injection point, spaced 1 cm apart across affected areas.
• Typical protocol: 3 monthly sessions, then a booster at 3–6 months.

• Inject intradermally into target areas (periorbital, cheeks, nasolabial folds) at 0.05–0.1 mL per point.
• Series of 3 sessions separated by 4 weeks yields better results than a single session.

• Ultrasound-guided injection directly into the tendon or joint.

Post-Procedure Care

• Apply ice packs to reduce swelling and discomfort.
• Avoid NSAIDs/aspirin for 1 week post-procedure.
• Avoid vigorous exercise for 24 hours.
• Mild erythema, swelling, and pain at injection sites are expected and typically resolve within a few days.

Classification Summary (Dohan Ehrenfest System)

Key Points

• ACD-A is the preferred anticoagulant for optimal platelet viability
• Double-spin method yields the highest platelet concentration
• Volume drawn: typically 10–30 mL for dermatologic use
• Activation is not required when injecting into vascularized soft tissue
• There is currently no universal standardized protocol — parameters vary across commercial kits and manual systems; reproducibility requires documenting your specific centrifuge RCF values