Objectives: Platelet Rich Plasma has widely been used in many branches of medicine for decades. Some groups believe in the mechanism of action of platelet rich plasma is promoting the healing cascade and some group does not. We believe in that the preparation of platelet rich plasma is crucial within the frame of clinical outcomes. Therefore, we demonstrated a study with non-anticoagulated prepared PRP protocols in order to evaluate the efficacy of the platelet rich plasma in-vitro with flow cytometry characterization, hypotonic stress, aggregometry and DEPA Classification.
Introduction: DEPA Classification is first proposed in 2016 by J. Magalon et al., in order to characterize and understand what exactly is injected into PRP patients. DEPA stands for Dose, Efficacy, Purity and Activation. It is considered to be the most important quality control measures and a full scale analysis of a prepared PRP to be injected into the patients. The cells were characterized using Flow Cytometry. Also, We performed the study with T0 and T4 timings for aggregation of platelets, hypotonic stress in order to identify whether any significant changes occured.
Materials / method: 4 different PRP was obtained. The NEXT PRP Syringes were centrifuged at 1500G X 4 mins. PPP was prepared in citrated tubes. In order to obtain PPP, 7 mins x 1000 RCF was performed. Upper 1,5 mL was harvested for evaluation. 0,5 mL PRP was measured with pH meter and CBC analyser at T0 and T4. In this study PRP samples were prepared without the use of any anticoagulant. Aggregometry was conducted via turbudimetric analysis on T0 and T4. Flow cytometry characterization was conducted blindly (also P-selectin) and spectrophotometric analysis was conducted. All PRP were calculated according to DEPA.
Results: Each samples were analysed due to DEPA classification. There were statistically significant differences between T0 and T4 levels of HSRT, and resting P-selectin levels The pH and aggregometry results were not statistically different. Some platelets were already activated before stimulation with ADP in all PRP product groups. This might be due to activation of platelets during preparation process.The level of already activated platelets were high in all PRP protocols before ADP stimulation at time point 4.
Conclusion: Further studies are needed to conclude whether the clinical outcome difference is significant between regular PRP and non-anticoagulated PRP. However, the Platelet Rich Plasma can also be obtained without the presence of any anticoagulant and the analysis can be done along with the data of total dose, efficacy of the protocol, purity and activation.
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