Influence of antiplatelet substances on platelet-rich plasma (original) (raw)
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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...
Archives of plastic surgery, 2013
Platelet-rich plasma (PRP) has more concentrated platelets than normal plasma (approximately 150-400×10(3) cell/dL). Platelets excrete several growth factors and cytokines that are associated with the healing and regeneration process. However, even though PRP is widely used, the mechanism or actual effect is presently unclear. Therefore, this study was performed to investigate the levels of growth factors and platelet concentration rate. Autologous blood for preparing PRP was obtained from healthy subjects aged 25 to 35 years. The samples were divided into 4 experimental groups (inactivated whole blood, inactivated PRP, activated whole blood with thrombin and calcium chloride, and activated PRP). The platelet counts in the blood were analyzed and the growth factors were quantitatively measured. A statistical analysis was performed by using Dunn's multiple comparison test. In the blood cell analysis, the platelet count of the PRP group was approximately 4.25 times higher than tha...
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.
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.
Cell and tissue banking, 2021
BACKGROUND The cellular and biochemical composition of the platelet rich plasma (PRP) may impact its regenerative capacity. PRP composition have been shown to vary substantially among different separation systems and protocols. The type and the dose of anticoagulant might affect the content of PRP. OBJECTIVE The objective of this study was to evaluate the effect of anticoagulant use, on cellular composition and the amount of growth factors in fresh PRP. METHODS Three different methods were used to prepare PRP from 12 healthy participants. The protocol 1 included standart dose sodium citrate (SC) (0.9 ml, 3.8%), protocol 2 included 0.5 ml SC and no anticoagulant was used in protocol 3. The PRP's were compared in regards to cellular content, capture efficiency of platelets (CE), concentrations and total doses of fresh studied vascular endothelial growth factor (VEGF), platelet derived growth factor -BB, (PDGF-BB), transforming growth factor β1 (TGF-β1) levels. RESULTS The CE and t...
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.
Basic studies on the clinical applications of platelet-rich plasma
Cell transplantation, 2003
Platelets, which contain many growth factors such as platelet-derived growth factor (PDGF) and transforming growth factor-beta (TGF-beta), are being used in clinical applications as platelet-rich plasma (PRP). Only a few studies, however, have been conducted on the growth factors present in PRP and on the clinical applications using the drug delivery system (DDS). For the purpose of clinical application, we first modified the PRP preparation method and assessed the amounts of growth factors contained in the human platelet concentrates. Furthermore, we assessed fibrin glue as a DDS of platelet concentrates. Platelet precipitations were made by twice centrifuging human whole blood. The precipitated platelet was resuspended to yield the platelet concentrates. The growth factor concentrations were measured. Fibrin glue sheets containing this platelet concentrate were implanted in rabbit pinna and samples were obtained for immunostaining (anti-PDGF antibody) to assess the use of PRP over...
INTRODUCTION: The diversity of procedures for obtaining platelet and plasmatic growth factors, the absence of control in most of them and the growing field of clinical application, makes them necessary methods adequately structured, documented, controlled and tested, playable by any author. The present series of clinical cases aims to introduce and test a specific technique for obtaining PRP, with precise characteristics both production and final composition of compound got, in 15 hematological healthy patients, comparing our results with those obtained by other procedures scientifically tested. MATERIAL AND METHODS: 15 caucasian patients were selected , 8 male and 7 female with age range between 35 and 65, healthy haematologically. The procedure for obtaining the PRP, consisted of a single centrifugation of the blood sample for 30 minutes at 3500 rpm in a angular shaft of 16 tubes centrifuge serie (CEMCON 2) and micropipetting the protein fraction rich in platelet and plasmatic growth factors and cell through open technique under aseptic conditions in horizontal laminar flow hood Grade A at a temperature of 22 ° C, with the use of leuco-platelet or Buffy-coat layer (PRP rich in leukocytes). RESULTS: No correlation between the amount of concentrated platelets and the amount of growth factors finally obtained was observed. The protocol set forth concentrated levels of platelets and leukocytes approximately 3 to 5 times higher than baseline levels with a predominance of mononuclear. Levels of growth factors from 7-10 times greater than the patient's baseline levels, with little variation in them. The growth factor levels were stable in the blood of each patient within 24 h of treatment between 7 and 9 times higher compared to the previous baseline. Compared with other procedures discussed in the literature; This method achieves concentration between 1.5 and 3 times more platelets in the final product, with a purification of growth factors overall type VEGF and TGF-B clearly superior. CONCLUSION: the technique disclosed is more effective since concentrate achieves greater amount of platelets and growth factors and efficient since it maintains a serum protein in these stable sera of patients after 24 hours of administration thereof.
Determination of platelet-derived growth factors in platelet unenriched plasma
Медицинский альманах, 2018
The authors describe a competitive aptamer based assay for detection of the platelet-derived growth factor BB (PDGF-BB; used as a model protein). The assay is making use of thrombin (a serine protease) as an enzyme label for reporting signals. It is taking advantage of a highly selective aptamer and of the fairly specific enzymatic activity of thrombin in terms of cleaving artificial fluorogenic peptide substrates. In a first step, the surface of wells of microplates is coated with PDGF-BB. On addition of a sample containing PDGF-BB, free and bound PDGF-BB compete with each other for binding to a DNA probe that consists of an aptamer sequence for PDGF-BB and a 29-mer aptamer sequence for thrombin. After washing, thrombin is added and will attach to the DNA probe that bound to the PDGF-BB on the microplates. Following addition of a fluorogenic peptide substrate, the bound thrombin will catalyze the cleavage of the substrate to generate a fluorescent product whose fluorescence intensity is measured at excitation/emission wavelengths of 370/ 440 nm. Fluorescence intensity decreases with increasing PDGF-BB concentration in the sample because less thrombin will bind to the PDGF-BB coated surface of the microplate. Under optimal conditions, PDGF-BB can be quantified in the 0.125 to 3 nM concentration range. This assay was successfully applied to the determination of PDGF-BB in spiked 100fold diluted human serum.