Low agreement between fresh and frozen‐thawed platelet‐rich plasma in the calibrated automated thrombogram assay

Ljungkvist, Marcus; Lövdahl, Susanna; Zetterberg, Eva; Berntorp, Erik

doi:10.1111/hae.13180

Cited by 9 publications

(9 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The supernatant (platelet-poor plasma) was collected and all platelet pellets were frozen at − 80 °C. Although differences have been observed in fresh versus frozen PRP [28], due to logistical concerns and because freezing has been demonstrated to be a valid method to induce the release of growth factors [20], frozen PL was used in this experiment in place of the fresh PRP used in the field.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of allogeneic freeze-dried platelet lysate in cartilage exposed to interleukin 1-β in vitro

2019

View full text Add to dashboard Cite

BackgroundPlatelet-rich plasma (PRP) as well as other platelet-derived products have been used as a potential disease-modifying treatment for musculoskeletal diseases, such as osteoarthritis (OA). The restorative properties of such products rely mainly on the high concentrations of growth factors, demonstrating encouraging results experimentally and clinically. Yet, the autologous blood-derived nature of the PRP product lead to limitations that precludes it’s widespread use. The main limitations for PRP use are; product variability, the need for minimum laboratory settings in most cases, and the need for storage at low temperatures to preserve its properties. Based on these limitations, the objective of this study was to investigate an allogeneic off-the-shelf platelet lysate (PL) in cartilage exposed to interleukin 1β (IL-1β). For this purpose, blood and cartilage were harvested from eight skeletally mature and healthy horses. Blood was processed into PL aliquots and divided into three groups (Frozen, Freeze-dried and Filtered freeze-dried), used in autologous and allogeneic conditions and in three different concentrations (1.5, 3 and 6-fold). Different PL preparations were then applied in cartilage culture with interleukin-1 beta and cultured for 10 days. Cartilage and media samples were collected and analyzed for total GAG and 35SO4-labeled GAG content.ResultsNo significant differences between the controls and PL groups in cartilage and media were demonstrated. The effects of PL on cartilage matrix were concentration dependent and intermediate concentrations (3-fold) in PL showed increased 35SO4-labelled GAG in cartilage.ConclusionIn conclusion, the allogeneic freeze-dried PL presented equivalent effects compared to frozen autologous PL. Intermediate platelet concentration on average demonstrated improved results, demonstrating less GAG loss compared to other concentrations.

show abstract

Section: Methodsmentioning

confidence: 99%

Evaluation of allogeneic freeze-dried platelet lysate in cartilage exposed to interleukin 1-β in vitro

2019

View full text Add to dashboard Cite

show abstract

“…Whereas TG results in frozen-thawed PRP were significantly higher compared to fresh PRP obtained from the same healthy subjects, several features including the detection of activated protein C resistance were comparable for the two sample conditions [55]. Recent work including healthy subjects and individuals at risk for bleeding showed poor agreement between TG results from fresh and frozen-thawed PRP [56]. Whether TG variations depend on individual functional characteristics of platelets that differ with the underlying pathophysiological conditions is not yet understood.…”

Section: Uncertainties and Limitations Of Thrombin Generation In The mentioning

confidence: 96%

Clinical Applications, Pitfalls, and Uncertainties of Thrombin Generation in the Presence of Platelets

Panova‐Noeva

Meijden

Cate

2019

JCM

View full text Add to dashboard Cite

Platelet-dependent thrombin generation is a helpful tool to assess ex vivo the interaction between platelets and plasma coagulation factors in the initiation, amplification, and inhibition of thrombin generation (TG). This review article discusses the most relevant available data on the clinical applications of fluorogenic TG, the most widely used TG assay, performed in the presence of platelets, i.e., in platelet-rich plasma. With respect to prothrombotic states, arterial hypertension and obesity were the most prominent cardiovascular conditions linked to increased platelet-dependent TG. In addition, platelet-associated hypercoagulability, assessed by the TG assay, has been shown in individuals with active cancer. In terms of bleeding, platelet-dependent TG has been applied to assess bleeding risk in individuals with hemophilia, von Willebrand disease, and Glanzmann thrombasthenia as well as in subjects with other congenital or acquired coagulation factor deficiencies. In addition to risk prediction, a role of the TG assay has been suggested in monitoring antiplatelet therapy in prothrombotic conditions and replacement therapy in bleeding diathesis. Finally, for the routine clinical use and as a biomarker of disease development and progression, better standardization and clinical validation of platelet-dependent TG are still needed.readings from a fluorometer are recorded and calculated by means of the Thrombinoscope software (Thrombinoscope BV). In principle, the measurements in PRP are assessed in fresh material, whereas TG in PFP is assessed in frozen, stored material. For TG measurements in PRP, coagulation is triggered by adding 20 µL exogenous tissue factor (TF) to 80 µL PRP (with adjusted platelet concentration of 150,000 platelets/µL), whereas for TG measurements in PFP, 20 µL exogenous TF and 4 µM phospholipids are added to 80 µL plasma. After prewarming for 10 min at 37 • C in the fluorimeter, the reaction is started by adding 20 µL of a low-affinity fluorogenic substrate for thrombin (Z-Gly-Gly-Arg-AMC) and calcium chloride mixture (FluCa). TG is measured as a function of a calibration signal obtained in a sample from the same plasma (PRP or PFP) after addition of a fixed amount of thrombin-α2-macroglobulin complex (20 µL of thrombin calibrator) and 20 µL of FluCa. The TG method is extensively reviewed in [5][6][7]. Parameters derived from a TG curve and most frequently reported in the studies are lag time, time to minimum thrombin formed expressed in minutes (min); peak height, maximum concentration of thrombin formed expressed in nM thrombin; and endogenous thrombin potential (ETP) or area under the curve expressed as nM of thrombin formed per minute.The aim of this article was to review the available evidence for the potential clinical application of platelet-dependent TG in both prothrombotic and bleeding conditions, as depicted in Figure 1. Aspects preventing its broader application and routine clinical practice will also be discussed as uncertainties and limitations of thrombin generation in the...

show abstract

“…A few groups have compared some of the TG assays in the past, and concluded that there was a poor correlation between the assays. 19,20 It is accepted that part of the poor correlation is due to lack of standardization.…”

Section: Introductionmentioning

confidence: 99%

“…Alternatively, the Innovance ETP assay (Siemens Healthcare Diagnostics), HaemoScan Thrombin Generation Assay (HemoScan), and Pefakit TDT (Pentapharm) are performed using a chromogenic substrate. A few groups have compared some of the TG assays in the past, and concluded that there was a poor correlation between the assays 19,20 . It is accepted that part of the poor correlation is due to lack of standardization.…”

Section: Introductionmentioning

confidence: 99%

Recommendations for the measurement of thrombin generation: Communication from the ISTH SSC Subcommittee on Lupus Anticoagulant/Antiphospholipid Antibodies

Ninivaggi

Laat-Kremers

Tripodi

et al. 2021

Journal of Thrombosis and Haemostasis

View full text Add to dashboard Cite

Low agreement between fresh and frozen‐thawed platelet‐rich plasma in the calibrated automated thrombogram assay

Abstract: Fresh and ft-PRP cannot be assumed to show equal results in the TGA-CAT assay.

Cited by 9 publications

References 18 publications

Evaluation of allogeneic freeze-dried platelet lysate in cartilage exposed to interleukin 1-β in vitro

Evaluation of allogeneic freeze-dried platelet lysate in cartilage exposed to interleukin 1-β in vitro

Clinical Applications, Pitfalls, and Uncertainties of Thrombin Generation in the Presence of Platelets

Recommendations for the measurement of thrombin generation: Communication from the ISTH SSC Subcommittee on Lupus Anticoagulant/Antiphospholipid Antibodies

Contact Info

Product

Resources

About