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Avoiding leukocyte contamination and early platelet activation in platelet-rich plasma
Ivy K Trindade-Suedam1, Fábio R M Leite, Juliana A N D de Morais
1Department of Diagnostic and Surgery, Division of Periodontics, School of Dentistry at Araraquara, São Paulo State University-UNESP, Brazil. ivytrin@uol.com.br
Abstract:
The objective of this study was to describe a new platelet-rich plasma (PRP) protocol with a reduced concentration of leukocytes and intact platelets. We collected 8 mL of venous blood (VB) from marginal ear veins of 10 male New Zealand white rabbits in acid dextrose citrate Vacutainer tubes. Tubes were centrifuged at 302g for 10 minutes. All plasma was collected in plastic tubes to avoid buffy-coat contamination and centrifuged at 2862g for 5 minutes. A 10% calcium chloride activator (10 PRP:2 CaCl2) was added to the lower third of this plasma (PRP), and the PRP gel was obtained. Mean platelet count was 317.7 x 10(3) +/- 39.9/microL in VB and 1344.9 x 10(3) +/- 347.5/microL in PRP. Leukocyte counts were 3.96 x 10(3) +/- 2.01/microL and 0.46 x 10(3) +/- 0.45/microL in VB and PRP, respectively. Mean platelet enrichment was 327.4 +/- 97.8%. All differences were statistically significant (P > .05). This protocol is practical and reproducible, resulting in a high concentration of intact platelets to help tissue repair and low levels of leukocytes.
Insights
This study introduces a new platelet-rich plasma (PRP) preparation method. The protocol yields a high concentration of intact platelets for tissue repair while significantly reducing leukocytes.
Area of Science:
- Regenerative Medicine
- Biotechnology
- Hematology
Background:
- Platelet-rich plasma (PRP) is utilized for tissue repair.
- Existing PRP protocols may have variable leukocyte concentrations.
- Optimizing PRP composition is crucial for therapeutic efficacy.
Purpose of the Study:
- To describe a novel platelet-rich plasma (PRP) protocol.
- To achieve a high concentration of intact platelets.
- To minimize leukocyte content in the final PRP product.
Main Methods:
- Venous blood (VB) collected from New Zealand white rabbits.
- Two-stage centrifugation process to isolate plasma and concentrate platelets.
- Addition of calcium chloride to activate and form PRP gel.
Main Results:
- Mean platelet count increased from 317.7 x 10(3)/microL in VB to 1344.9 x 10(3)/microL in PRP.
- Leukocyte count decreased from 3.96 x 10(3)/microL in VB to 0.46 x 10(3)/microL in PRP.
- Platelet enrichment achieved was 327.4%, with statistically significant differences (P > .05).
Conclusions:
- The described PRP protocol is practical and reproducible.
- It effectively produces PRP with a high concentration of intact platelets.
- The protocol yields low levels of leukocytes, potentially enhancing therapeutic applications.
