Related Experiment Video
Updated: Jun 10, 2025
Turbidimetry on Human Washed Platelets: The Effect of the Pannexin1-inhibitor Brilliant Blue FCF on Collagen-induced Aggregation
Published on: April 6, 2017
Optimized Methodology to Produce Platelet-Rich Plasma and Perform Platelet Aggregation in Patients With Coronary
Atul K Kashyap1, Bonu Saikrishna1, Bhanu Duggal1
1Cardiology, All India Institute of Medical Sciences, Rishikesh, Rishikesh, IND.
Insights
Optimized platelet-rich plasma (PRP) preparation using separating gel tubes for light transmittance aggregometry (LTA) in coronary artery disease (CAD) patients. This method reliably measures platelet aggregation (PA) in patients on clopidogrel and ticagrelor.
Area of Science:
- Cardiovascular Medicine
- Hematology
- Clinical Chemistry
Background:
- Inter-individual variability in clopidogrel metabolism and platelet counts necessitates optimized assays.
- Coronary artery disease (CAD) patients on antiplatelet therapy require accurate monitoring of platelet function.
- Light transmittance aggregometry (LTA) is a key method for assessing platelet aggregation (PA).
Purpose of the Study:
- To optimize platelet-rich plasma (PRP) preparation for LTA in CAD patients.
- To evaluate the efficacy of separating gel PRP tubes for routine LTA analysis.
- To assess PA in CAD patients treated with clopidogrel and ticagrelor.
Main Methods:
- PRP and platelet-poor plasma (PPP) were prepared using separating gel PRP tubes and standard centrifugation protocols.
- LTA was performed using adenosine diphosphate (ADP) as an agonist on healthy controls and CAD patients.
- The study involved 10 healthy controls and 28 CAD patients (16 on clopidogrel, 12 on ticagrelor).
Main Results:
- Optimized PRP preparation using PRP tubes was achieved.
- Mean maximum platelet aggregation (MPA) differed significantly between patient groups (p=0.042).
- Mean platelet volumes showed significant differences across groups (p≤0.01).
Conclusions:
- Separating gel PRP tubes provide a reliable method for PRP preparation for LTA.
- The optimized LTA assay is suitable for routine measurement of PA in cardiology clinics.
- This method aids in managing antiplatelet therapy in CAD patients.
Background:
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.
Methodology:
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.
Results:
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±2.51 fL, 7.60±0.85 fL, and 10.99±1.62 fL (p≤0.01), respectively.
Conclusion:
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.
