Deaprtment of Transfusion Medicine ,Postgraduate Institute of Medical Education and Research CHANDIGARH, Chandigarh, India
Background/Case Studies: Storage of platelets remains a challenge due to morphological, functional, and biochemical changes that compromise efficacy and safety of platelet transfusion
Study
Design/Methods: A collaborative study was planned by the Departments of Transfusion Medicine, Haematology, Biochemistry, and Histopathology of Postgraduate Institute of Medical Education and Research, Chandigarh, India. After ethics approval, forty-two healthy blood donors (free from anti-platelet drugs for 1 week) were recruited. Whole blood was collected in 450ml CPDA-1 blood Collection bags. Buffy Coat derived Platelet-Concentrate was prepared and stored at 22°C in an agitator. On day 1, four platelet aliquots were prepared: 1st as baseline, 2nd supplemented with 10 µM resveratrol, 3rd with 50 µg/ml curcumin and the 4th acted as standard control. Baseline parameters were analysed on day 1, and other three aliquots were analysed on day 5 of storage. Morphology was evaluated via Transmission Electron Microscopy (TEM) and scored using a pre-defined scoring scale of 0 (unchanged discoid shape platelets) to 3 (degenerated/necrotic platelets). Platelet functions were assessed via light transmission aggregometry using ADP, Arachidonic Acid, collagen & epinephrine as agonists. Reactive Oxygen Species (ROS) production was quantified with 2,7-dichlorofluorescein diacetate dye, apoptosis via Annexin-V, and P-selectin expression using an APC-conjugated monoclonal antibody through flow-cytometry.
Results/Findings: On day 5 of storage, resveratrol and curcumin treated platelets exhibited a significantly higher (P< 0.05) platelet count (560.1 x103/µl and 549.42 x103/µl) as compared to control platelets (458.7.1 x103/µl). TEM revealed better structural preservation (Score 2) in platelets in resveratrol and Curcumin, than control group (Score 3). Platelet functions were better preserved in antioxidant solutions as compared to controls, with the aggregation studies (P< 0.05) showing a response of ADP (50.7%, 47.05% vs. 37.5%), epinephrine (52.9%, 51.16% vs. 40.8%), collagen (39.6%, 37.4% vs. 27.4%) and arachidonic acid (47.2%, 45.35% vs.37.08%). Similarly, platelets in resveratrol & curcumin showed significantly lower expression (P< 0.05) of Annexin-V (51.9%, 50.9%), P-selectin (31.7%, 28.3%) and intracellular ROS generation (MFI: 444.89, 370.2), than control group (Annexin-V: 63.79%, P-selectin: 35.24%, ROS: 697.89 MFI).
Conclusions: Conclusion: Our findings indicate that, resveratrol and curcumin better preserve platelet function and structure in comparison to the currently available storage solutions. Hence, they can be promising ingredients of new platelet additive solutions.
