Optimized Methodology to Produce Platelet-Rich Plasma and Perform Platelet Aggregation in Patients With Coronary Artery Disease.

Abstract

Inter-individual differences in clopidogrel metabolism and platelet counts within platelet-rich plasma (PRP) intrigued us to optimize light transmittance aggregometry (LTA) assay in coronary artery disease (CAD) patients administered with clopidogrel and ticagrelor.The objective of the study was to optimize PRP preparation using separating gel PRP tubes to perform LTA among CAD patients on clopidogrel and ticagrelor. Initially, we optimized PRP preparation and platelet aggregation (PA) on healthy controls. To validate the protocol, we recruited 10 healthy controls and 28 CAD patients, comprising 16 on clopidogrel and 12 on ticagrelor regimen. Bio-X, India, supplied PRP tubes (9 mL) with 3.2% sodium citrate. PRP and autologous platelet-poor plasma (PPP) were prepared by centrifugation at 151 g for seven minutes and 3,780 g for 10 minutes, respectively. LTA was performed using platelet aggregometer TA-4V (Stago, Asnières-sur-Seine, France). Adenosine diphosphate (ADP) (10 µM) was used as an agonist. The mean maximum platelet aggregation (MPA) among controls, clopidogrel patients, and ticagrelor patients were 55.161±13.69%,53.17±22%, and 35.84±20.79% (p=0.042), respectively. The mean platelet volumes among groups were 9.34±2.51fL, 7.60±0.85fL, and 10.99±1.62fL (p≤0.01), respectively. We successfully optimized PRP preparation for the LTA assay using a PRP tube. Its application to measure PA routinely in a cardiology clinic seems propitious.