Basic studies on the clinical applications of platelet-rich plasma (original) (raw)

Determining the Effect of Preparation and Storage: An Effort to Streamline Platelet Components as a Source of Growth Factors for Clinical Application

Transfusion Medicine and Hemotherapy, 2015

Background: In the present study, different methods for preparation of platelet-rich plasma (PRP) are investigated in order to standardize the component in terms of growth factor content. The effects of concentration technique and storage duration are also analyzed. Methods: PRP was collected from 40 donors by plateletpheresis as well as by the buffy coat and tube method. Concentration of growth factors was performed using double freeze thaw- and CaCl2-induced degranulation techniques. Growth factor estimation was performed using ELISA. Results: The levels of growth factors were highest in PRP from buffy coat, moderately lower in plasma gained by plateletpheresis and lowest in that obtained by the tube method. Mean levels of platelet-derived growth factors (PDGF) AB and BB are significantly higher when CaCl2 was used for concentrating the growth factors. The mean levels of transforming growth factor β1 and insulin-like growth factor I were higher when applying the double freeze thaw...

Do the Fibrin Architecture and Leukocyte Content Influence the Growth Factor Release of Platelet Concentrates? An Evidence-based Answer Comparing a Pure Platelet-Rich Plasma (P-PRP) Gel and a Leukocyte- and Platelet-Rich Fibrin (L-PRF)

Current Pharmaceutical Biotechnology, 2012

Platelet concentrates for surgical use are tools of regenerative medicine designed for the local release of platelet growth factors into a surgical or wounded site, in order to stimulate tissue healing or regeneration. Leukocyte content and fibrin architecture are 2 key characteristics of all platelet concentrates and allow to classify these technologies in 4 families, but very little is known about the impact of these 2 parameters on the intrinsic biology of these products. In this demonstration, we highlight some outstanding differences in the growth factor and matrix protein release between 2 families of platelet concentrate: Pure Platelet-Rich Plasma (P-PRP, here the Anitua's PRGF-Preparation Rich in Growth Factorstechnique) and Leukocyte-and Platelet-Rich Fibrin (L-PRF, here the Choukroun's method). These 2 families are the extreme opposites in terms of fibrin architecture and leukocyte content. The slow release of 3 key growth factors (Transforming Growth Factor 1 (TGF 1), Platelet-Derived Growth Factor AB (PDGF-AB) and Vascular Endothelial Growth Factor (VEGF)) and matrix proteins (fibronectin, vitronectin and thrombospondin-1) from the L-PRF and P-PRP gel membranes in culture medium is described and discussed. During 7 days, the L-PRF membranes slowly release significantly larger amounts of all these molecules than the P-PRP gel membranes, and the 2 products display different release patterns. In both platelet concentrates, vitronectin is the sole molecule to be released almost completely after only 4 hours, suggesting that this molecule is not trapped in the fibrin matrix and not produced by the leukocytes. Moreover the P-PRP gel membranes completely dissolve in the culture medium after less than 5 days only, while the L-PRF membranes are still intact after 7 days. This simple demonstration shows that the polymerization and final architecture of the fibrin matrix considerably influence the strength and the growth factor trapping/release potential of the membrane. It also suggests that the leukocyte populations have a strong influence on the release of some growth factors, particularly TGF 1. Finally, the various platelet concentrates present very different biological characteristics, and an accurate definition and characterization of the different families of product is a key issue for a better understanding and comparison of the reported clinical effects of these surgical adjuvants.

In vitro release of growth factors from platelet-rich fibrin (PRF): a proposal to optimize the clinical applications of PRF

2009

Objective. Determine the release of growth factors (GF) from platelet-rich fibrin (PRF) and supernatant serum to optimize clinical use. Study design. Platelet-derived growth factors-AB (PDGF-AB), transforming growth factor-␤1 (TGF-␤1), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), and insulin-like growth factor-1 (IGF-1) were quantified in PRF releasate and in the supernatant serum (N ϭ 8) over 300 minutes after clot formation. Protein profiles were determined by SDS-PAGE. Results. Mean quantity of PDGF-AB, TGF-ß1, VEGF, and EGF in PRF releasate increased significantly to about 52, 72, 1, and 3 ng, respectively, whereas mean IGF-1 content remained at 250 ng. GF was also found in serum supernatant. Protein profiles of the releasates and the supernatant serum were similar. Conclusion. The PRF membrane should be used immediately after formation to maximize release of GF to the surgical site. The remaining fluid can be recovered as an additional source of GF for grafting.

Comparison of Growth Factor and Platelet Concentration From Commercial Platelet-Rich Plasma Separation Systems

American Journal of Sports Medicine, 2010

Background: Clinical studies claim that platelet-rich plasma (PRP) shortens recovery times because of its high concentration of growth factors that may enhance the tissue repair process. Most of these studies obtained PRP using different separation systems, and few analyzed the content of the PRP used as treatment. Purpose: This study characterized the composition of single-donor PRP produced by 3 commercially available PRP separation systems. Study Design: Controlled laboratory study. Methods: Five healthy humans donated 100 mL of blood, which was processed to produce PRP using 3 PRP concentration systems (MTF Cascade, Arteriocyte Magellan, Biomet GPS III). Platelet, white blood cell (WBC), red blood cell, and fibrinogen concentrations were analyzed by automated systems in a clinical laboratory, whereas ELISA determined the concentrations of platelet-derived growth factor ab and bb (PDGF-ab, PDGF-bb), transforming growth factor b1 (TGF-b1), and vascular endothelial growth factor (VEGF). Results: There was no significant difference in mean PRP platelet, red blood cell, active TGF-b1, or fibrinogen concentrations among PRP separation systems. There was a significant difference in platelet capture efficiency. The highest platelet capture efficiency was obtained with Cascade, which was comparable with Magellan but significantly higher than GPS III. There was a significant difference among all systems in the concentrations of WBC, PDGF-ab, PDGF-bb, and VEGF. The Cascade system concentrated leukocyte-poor PRP, compared with leukocyte-rich PRP from the GPS III and Magellan systems. Conclusion: The GPS III and Magellan concentrate leukocyte-rich PRP, which results in increased concentrations of WBCs, PDGF-ab, PDGF-bb, and VEGF as compared with the leukocyte-poor PRP from Cascade. Overall, there was no significant difference among systems in the platelet concentration, red blood cell, active TGF-b1, or fibrinogen levels. Clinical Relevance: Products from commercially available PRP separation systems produce differing concentrations of growth factors and WBCs. Further research is necessary to determine the clinical relevance of these findings.

Platelet and growth factor concentrations in activated platelet-rich plasma: a comparison of seven commercial separation systems

Journal of Artificial Organs, 2014

Platelet-rich plasma (PRP) is blood plasma that has been enriched with platelets. It holds promise for clinical use in areas such as wound healing and regenerative medicine, including bone regeneration. This study characterized the composition of PRP produced by seven commercially available separation systems (JP200, GLO PRP, Magellan Autologous Platelet Separator System, KYOCERA Medical PRP Kit, SELPHYL, MyCells, and Dr. Shin's System THROMBO KIT) to evaluate the platelet, white blood cell, red blood cell, and growth factor concentrations, as well as platelet-derived growth factor-AB (PDGF-AB), transforming growth factor beta-1 (TGF-b1), and vascular endothelial growth factor (VEGF) concentrations. PRP prepared using the Magellan Autologous Platelet Separator System and the KYOCERA Medical PRP Kit contained the highest platelet concentrations. The mean PDGF-AB concentration of activated PRP was the highest from JP200, followed by the KYOCERA Medical PRP Kit, Magellan Autologous Platelet Separator System, MyCells, and GLO PRP. TGF-b1 and VEGF concentrations varied greatly among individual samples, and there was almost no significant difference among the different systems, unlike for PDGF. The SELPHYL system produced PRP with low concentrations of both platelets and growth factors. Commercial PRP separation systems vary widely, and familiarity with their individual advantages is important to extend their clinical application to a wide variety of conditions.

Platelet content and growth factor release in platelet-rich plasma: a comparison of four different systems

Vox Sanguinis, 2006

BACKGROUND: Cell therapy and cell culture have received much attention in recent decades. Suitable cell growth requires growth supplements such as fetal bovine serum (FBS). FBS is component rich in nutrients, growth factors and supplementary compounds. However, FBS utilization has some limitations including mass production. Therefore, finding alternatives with the same growth promoting effects is inevitable. OBJECTIVES: This study was designed to compare the effect of bovine platelet lysate (PL) and PRP on different cell lines as a cost effective and available alternative for FBS. METHODS: Three conventional cell lines were investigated. Protein pattern of PL and platelet rich plasma (PRP) in comparison to FBS was determined using SDS page electrophoresis, and MTT and plating efficiency of cell lines in presence of PL and PRP were evaluated. RESULTS: The results demonstrated that platelet rich plasma and platelet lysate could increase cells' viability similar to FBS. These results were significant in comparison with control group. CONCLUSIONS: It can be concluded that platelet lysate could be a valuable candidate to replace FBS in cell culture techniques, however, more studies should be done to understand its exact efficacy.