Determining the Effect of Preparation and Storage: An Effort to Streamline Platelet Components as a Source of Growth Factors for Clinical Application (original) (raw)
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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.
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.
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 work aims to introduce and test a specific technique for obtaining PRP, with precise characteristics both production and final composition of compound got, in 350 hematological healthy patients, comparing our results with those obtained by other procedures scientifically tested. Material and Methods: 350 caucasian patients were selected, 175 male and 175 female with age range between 15 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 Conclusion: Compared with other procedures discussed in the literature; This method achieves concentration between 1.5 and 3 times more platelets in the final product, as we can see in figure 5, with a purification of growth factors overall type VEGF and TGF-B clearly superior, visible in figure 6.
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.
Journal of Experimental Orthopaedics, 2019
Background: Alternative methods of platelet-rich plasma (PRP) preparation, storage, and activation that can be stably reproduced are needed to improve PRP production. The purpose of this study was to investigate the effect of the preparer's experience on the quality of prepared PRP, chronological changes occurring in PRP, and the effect of the activation procedures on the release of several growth factors from PRP, using PRP prepared with the PRGF-Endoret Kit. Methods: Leukocyte-poor PRP samples from seventeen healthy volunteers were prepared using the PRGF-Endoret Kit and the PRGF IV System Centrifuge. The platelet and leukocyte concentrations were compared based on the preparer's experience. The concentrations of platelets, hepatocyte growth factor (HGF), platelet-derived growth factor-BB (PDGF-BB), and insulin-like growth factor-1 (IGF-1) were determined at 0 and 60 min after PRP preparation, and compared. Concentrations of the above growth factors from PRP activated by freeze-thaw cycling and by calcium chloride (CaCl 2) were also compared. Results: No significant difference was observed in the platelet concentrations and leukocyte contamination rates, based on the preparer's experience. At 60 min after PRP preparation, the platelet concentration decreased significantly, while the HGF, PDGF-BB, and IGF-1 concentrations remained unchanged. Activation with CaCl 2 resulted in a significant increase in the PDGF-BB levels, although the HGF and IGF-1 concentrations remained unchanged. Conclusions: The results of this study show that leukocyte-poor PRP prepared using the PRGF-Endoret Kit did not result in any qualitative difference that depended on the experience of the preparer. However, PRP preparation required standardization in terms of the time of blood count measurement. Growth factor concentrations in PRP differed according to the platelet-activation method used.
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.
International Journal of PharmTech Research
Platelet concentrate (PC) has been used as substitute to the use of fetal bovine serum (FBS) in cell culture media. However, it's use as additive in cell culture media showed inconsistent results on cell proliferation, and the inconsistent results may be due to variability of platelet counts or growth factor content. Standard protocols are lacking for the preparation of PC before it is applied as additive in cell culture media. The growth factor content of PC can be released by freeze-thaw cycles, which range from one to three cycles before use for cell culture media. This study aimed to measure base-line platelet counts and growth factor levels and compare to platelet counts and growth factor levels after one, two and three freeze-thaw cycles. In this study, we obtained PC from Indonesian Red Cross. The PCs were aliquoted and stored at-20 o C and then subjected to 1 to 3 freeze-thaw cycles. The number of platelet before and after freeze-thaw cycles were measured using Sysmex XN...
Growth Factors, 2006
Background: In this study, three commercial systems for the preparation of platelet-rich plasma (PRP) were compared and platelet growth factors release was measured. Methods: Ten healthy volunteers donated whole blood that was fractionated by a blood cell separator, and a table-top centrifuge to prepare PRP. Furthermore, an autologous growth factor filter was used to concentrate PRP fractionated by the blood cell separator. PRP was subsequently activated with autologously produced thrombin to degranulate the platelets to measure platelet-derived growth factor-AB (PDGF-AB), transforming growth factor-beta (TGF-b), insulin-like growth factor-1 (IGF-1), and vascular endothelial growth factor (VEGF). Results: PRP contained significantly higher platelet counts compared with baseline values (p , 0.001). PDGF-AB concentrations were increased more than 18-fold in the platelet gel supernatant when the cell-separator and GPS were used, whereas only a 3-fold increase was seen with the AGF. Conclusion: The three PRP devices enable the preparation of PRP for the release of high concentrations of platelet growth factor, but showed different harvesting capacities for the collection of concentrated platelets. The administration of thrombin for PRP activation resulted in the release of high concentrations of PDGF-AB and TGF-b but only when PRP had not been activated during the preparation process in vitro.