A novel method for harvesting concentrated platelet-rich fibrin (C-PRF) with a 10-fold increase in platelet and leukocyte yields

Abstract

Background and objectives Liquid platelet rich fibrin (PRF; often referred to as injectable PRF) has been utilized as an injectable formulation of PRF that is capable of stimulating tissue regeneration. Our research group recently found that following standard L-PRF protocols (2700 RPM for 12 min), a massive increase in platelets and leukocytes was observed directly within the buffycoat layer directly above the red blood cell layer. The purpose of this study was to develop a novel harvesting technique to isolate liquid PRF directly from this buffy coat layer and to compare this technique to standard i-PRF. Materials and methods Standard high g-force L-PRF and low g-force i-PRF protocols were utilized to separate blood layers. Above the red blood corpuscle layer, sequential 100-μL layers of plasma were harvested (12 layers total; i.e., 1.2 mL, which represents the total i-PRF volume), and 3 layers (3 × 100 μL) were harvested from the red blood cell layer to quantify blood cells. Each layer was then sent for complete blood count (CBC) analysis, and the cell numbers were quantified including red blood cells, leukocytes, neutrophils, lymphocytes, monocytes, and platelets. The liquid PRF that was directly collected from the buffycoat layer following L-PRF protocols was referred to as concentrated PRF (C-PRF).Results The i-PRF protocol typically yielded a 2-to 3-fold increase in platelets and a l.5-fold increase in leukocyte concentration from the 1-to 1.2-mL plasma layer compared to baseline concentrations in whole blood. While almost no cells were found in the first 4-mL layer of L-PRF, a massive accumulation of platelets and leukocytes was found directly within the buffy coat layer demonstrating extremely high concentrations of cells in this 0.3-0.5-mL layer (~20-fold increases). We therefore proposed harvesting this 0.3-to 0.5-mL layer directly above the red blood cell corpuscle layer as liquid C-PRF. In general, i-PRF was able to increase platelet numbers by ~250%, whereas a 1200-1700% increase in platelet numbers could easily be achieved by harvesting this 0.3-0.5 mL of C-PRF (total platelet concentrations of > 2000-3000 × 10 9 cells/L). Conclusion While conventional i-PRF protocols increase platelet yield by 2-3-fold and leukocyte yield by 50%, we convincingly demonstrated the ability to concentrate platelets and leukocytes over 10-fold by harvesting the 0.3-0.5 mL of C-PRF within the buffy coat following L-PRF protocols. Clinical relevance Previous studies have demonstrated only a slight increase in platelet and leukocyte concentrations in i-PRF. The present study described a novel harvesting technique with over a 10-fold increase in platelets and leukocytes that can be further utilized for tissue regeneration.

Quantitative Near-Infrared Imaging of Platelets in Platelet-Rich Fibrin (PRF) Matrices: Comparative Analysis of Bio-PRF, Leukocyte-Rich PRF, Advanced-PRF and Concentrated Growth Factors

“…In recent years, efforts to modify the original Chouroun’s method for leukocyte and platelet-rich fibrin (L-PRF) [ 8 ] have led to the development of several PRF derivatives, including concentrated growth factors (CGF) [ 9 ] and advanced-PRF (A-PRF) [ 6 ]. More recently, this has led to the development of another PRF derivative, bio-PRF [ 10 ]. According to our understanding, the main goals of these modifications have been to: (1) form mechanically tough fibrin matrices using thicker and well-crosslinked fibrin fibers; (2) improve the capacity for growth factor retention and release; (3) exclude (or include) more leukocytes.…”

Liquid Platelet-Rich Fibrin and Heat-Coagulated Albumin Gel: Bioassays for TGF-β Activity

“…After cooling, the PPP gel is mixed back with the original buffy coat fraction that maintained the coagulating properties. The liquid buffy coat is also termed concentrated PRF or C-PRF [11]. Thus, the viscous injectable PRF mixture consists of a gel prepared from heated PPP and the non-heated liquid buffy coat C-PRF fraction (Alb-PRF).…”

The Platelet Concentrates Therapy: From the Biased Past to the Anticipated Future

“…In the third generation, much attention was paid to the comparisons between PRP derivatives. In particular, the ability of PRP to retain and release growth factors and the distribution of platelets and leukocytes have been vigorously investigated [30][31][32][33][34]. It is essential and necessary to compare individual PCs from a neutral standpoint in order to optimize and standardize the preparation protocol.…”