Richard J Miron1,2, Jihua Chai3, Peng Zhang3, Yuqing Li3, Yunxiao Wang3, Carlos Fernando de Almeida Barros Mourão4, Anton Sculean5, Masako Fujioka Kobayashi6, Yufeng Zhang3,7. 1. The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, Wuhan, China. richard.miron@zmk.unibe.ch. 2. Department of Periodontology, University of Bern, Bern, Switzerland. richard.miron@zmk.unibe.ch. 3. The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, Wuhan, China. 4. Department of Oral Surgery, Dentistry School, Fluminense Federal University, Niterói, Rio de Janeiro, Brazil. 5. Department of Periodontology, University of Bern, Bern, Switzerland. 6. Department of Maxillofacial Surgery, Inselspital, University of Bern, Bern, Switzerland. 7. Department of Dental Implantology, School and Hospital of Stomatology, Wuhan University, Wuhan, China.
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 buffy-coat 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 buffy-coat 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 × 109 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.
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 buffy-coat 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 buffy-coat 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 × 109 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.