Vitamin C mitigates oxidative stress and preserves platelet efficacy during prolonged storage ‒ an in vitro study
DOI:
https://doi.org/10.15584/ejcem.2026.3.7Keywords:
antioxidants, blood platelets, oxidative stress, platelet banking, vitamin CAbstract
Introduction and aim. Platelets stored at 22–24°C beyond 3–5 days undergo oxidative changes leading to storage lesions (PSL). Platelet additive solutions (PAS) improve platelet shelf-life, and antioxidant additives in PAS can mitigate PSL. This in vitro study evaluated the impact of Vitamin C (VC), a potent free radical scavenger, as an antioxidant additive in PAS-SSP+ on the redox status and functional integrity of rat platelets during prolonged storage.
Material and methods. Platelets from male Wistar rats (n=5) were grouped into controls (SSP+) and 10-VC (10mM VC in SSP+) and stored at 22–24°C for 11 days. Markers of antioxidant and oxidative status, platelet function and metabolism were investigated on days 1, 4, 7 and 11.
Results. Superoxide radicals, thiobarbituric acid reactive substances, and advanced oxidation protein products were significantly lower in 10-VC than controls. Superoxide dismutase and catalase were maintained whereas, glutathione was higher by 27% (p<0.01) and protein sulfhydryls by 65% (p<0.01) on day 7 than controls. Total antioxidant capacity was significantly higher than controls throughout storage in 10-VC. P-selectin lowered by 58% (p<0.001) on day 7 than controls while aggregation and cell viability were preserved in 10-VC.
Conclusion. 10-VC reduced oxidative stress by scavenging reactive species, inhibiting lipid and protein oxidation, augmenting antioxidant capacity, and preserving platelet functions and viability. Thus, VC in SSP+ maintained redox status and improved platelet efficacy, highlighting its potential as an additive in PAS ‘for preserving platelet integrity during prolonged storage.
Downloads
References
Rajashekaraiah V, Rajanand MC. Platelet storage: Progress so far. J Thromb Thrombolysis. 2023;55(1):9-17. doi:10.1007/s11239-022-02716-3
Shrivastava M. The platelet storage lesion. Transfus Apher Sci. 2009;41(2):105-113. doi:10.1016/j.transci.2009.07.002
Gulliksson H. Additive solutions for the storage of platelets for transfusion. Transfus Med. 2000;10(4):257-264. doi:10.1046/j.1365-3148.2000.00262.x
Ringwald J, Zimmermann R, Eckstein R. The new generation of platelet additive solution for storage at 22 C: development and current experience. Transfus Med Rev. 2006;20(2):158-164. doi:10.1016/j.tmrv.2005.11.003
Hornsey VS, McColl K, Drummond O, et al. Extended storage of platelets in SSP+ platelet additive solution. Vox Sang. 2006;91(1):41-46. doi:10.1111/j.1423-0410.2006.00771.x
Saunders C, Rowe G, Wilkins K, Holme S, Collins P. In vitro storage characteristics of platelet concentrates suspended in 70% SSP+ TM additive solution versus plasma over a 14‐day storage period. Vox Sang. 2011;101(2):112-121. doi:10.1111/j.1423-0410.2011.01468.x
Alexandru N, Popov D, Georgescu A. Intraplatelet oxidative/nitrative stress: inductors, consequences, and control. Trends Cardiovas Med. 2010;20(7):232-238. doi:10.1016/j.tcm.2011.11.007
Rinalducci S, Zolla L. Biochemistry of storage lesions of red cell and platelet concentrates: A continuous fight implying oxidative/nitrosative/phosphorylative stress and signaling. Transfus Apher Sci. 2015;52(3):262-269. doi:10.1016/j.transci.2015.04.005
Arthur JF, Shen Y, Kahn ML, Berndt MC, Andrews RK, Gardiner EE. Ligand binding rapidly induces disulfide-dependent dimerization of glycoprotein VI on the platelet plasma membrane. J Biol Chem. 2007;282(42):30434-30441. doi:10.1074/jbc.M701330200
Qiao J, Arthur JF, Gardiner EE, Andrews RK, Zeng L, Xu K. Regulation of platelet activation and thrombus formation by reactive oxygen species. Redox Biol. 2018;14:126-130. doi:10.1016/j.redox.2017.08.021
Ray PD, Huang BW, Tsuji Y. Reactive oxygen species (ROS) homeostasis and redox regulation in cellular signaling. Cell Signal. 2012;24(5):981-990. doi:10.1016/j.cellsig.2012.01.008
Zhuang Y, Ren G, Li H, et al. In vitro properties of apheresis platelet during extended storage in plasma treated with anandamide. Transfus Apher Sci. 2014;51(1):58-64. doi:10.1016/j.transci.2014.03.009
Wang X, Fan Y, Shi R, Li J, Zhao S. Quality assessment of platelets stored in a modified platelet additive solution with trehalose at low temperature (10C) and in vivo effects on rabbit model of thrombocytopenia. Platelets. 2015;26(1):72-79. doi:10.3109/09537104.2013.872772
Handigund M, Bae TW, Lee J, Cho YG. Evaluation of in vitro storage characteristics of cold stored platelet concentrates with N acetylcysteine (NAC). Transfus Apher Sci. 2016;54(1):127-138. doi:10.1016/j.transci.2016.01.006
Mithun M, Rajashekaraiah. L-Carnitine as an additive in Tyrode's buffer during platelet storage. Blood Coagul Fibrinolysis. 2018;29(7):613-621. doi:10.1097/MBC.0000000000000760
Ekaney ML, Gray GG, McKillop IH, Evans SL. Enhanced platelet function in cold stored whole blood supplemented with resveratrol or cytochrome C. J Trauma Acute Care Surg. 2018;85(1):92-97. doi:10.1097/TA.0000000000001887
Hosseini E, Ghasemzadeh M, Atashibarg M, Haghshenas M. ROS scavenger, N‐acetyl‐l‐cysteine and NOX specific inhibitor, VAS2870 reduce platelets apoptosis while enhancing their viability during storage. Transfus. 2019;59(4):1333-1343. doi:10.1111/ trf.15114
Wang L, Xie R, Fan Z, et al. The contribution of oxidative stress to platelet senescence during storage. Transfus 2019; 59(7):2389-2402. doi:10.1111/trf.15291
Amiri F, Dahaj MM, Siasi NH, Deyhim MR. Treatment of platelet concentrates with the L-carnitine modulates platelets oxidative stress and platelet apoptosis due to mitochondrial reactive oxygen species reduction and reducing cytochrome C release during storage. J Thromb Thrombolysis. 2021;51(2):277-285. doi:10.1007/s11239-020-02241-1
Hegde S, Wellendorf AM, Zheng Y, Cancelas JA. Antioxidant prevents clearance of hemostatically competent platelets after long‐term cold storage. Transfus. 2021;61(2):557-567. doi:10.1111/trf.16200
Zhang P, Du J, Zhao L, et al. The role of intraplatelet reactive oxygen species in the regulation of platelet glycoprotein Ibα ectodomain shedding. Thromb Res. 2013:132(6):696-701. doi:10.1016/j.thromres.2013.09.034
Stef G, Csiszar A, Xiangmin Z, Ferdinandy P, Ungvari Z, Veress G. Inhibition of NAD (P) H oxidase attenuates aggregation of platelets from high-risk cardiac patients with aspirin resistance. Pharmacol Rep. 2007;59(4):428. PMID: 17901572
Rajanand MC, Ananthakrishna AB, Rajashekaraiah V. A laboratory study on N-acetyl cysteine in SSP+ modulating oxidative stress and delaying the progression of storage lesion in platelets. Biomed Res Ther. 2024;11(11):6950-6959. doi:0.15419/bmrat.v11i11.941
Rajanand MC, Ananthakrishna AB, Rajashekaraiah V. Caffeic Acid Alleviates Oxidative Stress and Prolongs the Shelf-Life of Rat Platelets. Biopreserv Biobank. 2026;24(1):41-51. doi:10.1089/bio.2025.0039
Olas B, Wachowicz B, Buczyn A. Vitamin C suppresses the cisplatin toxicity on blood platelets. Anti-cancer drugs. 2000;11(6):487-493. doi:10.1097/00001813-200007000-00010
Olas B, Wachowicz B. Resveratrol and vitamin C as antioxidants in blood platelets. Thromb Res. 2002;106(2):143-148. doi:10.1016/S0049-3848(02)00101-9
Halliwell B, Gutteridge JMC. Free Radicals in Biology and Medicine, 5th ed. New York: Oxford University Press; 2015.
Pignatelli P, Sanguigni V, Paola SG, Coco EL, Lenti L, Violi F. Vitamin C inhibits platelet expression of CD40 ligand. Free Rad Biol Med. 2005;38(12):1662-1666. doi:10.1016/j.freeradbiomed.2005.02.032
Mohammed BM, Sanford KW, Fisher BJ, et al. Impact of high dose vitamin C on platelet function. World J Crit Care Med. 2017;6(1):37. doi:10.5492/wjccm.v6.i1.37
Rajanand MC, Ananthakrishna AB, Rajashekaraiah V. Oxidative modulations in platelets stored in SSP+, PAS-G and Tyrode's buffer: a comparative analysis. Hematol Transfus Cell Ther. 2024;46:80-89. doi:10.1016/j.htct.2024.04.121
Blasa M, Angelino D, Gennari L, Ninfali P. The cellular antioxidant activity in red blood cells (CAA-RBC): a new approach to bioavailability and synergy of phytochemicals and botanical extracts. Food Chem. 2011;125(2):685-691. doi:10.1016/j.foodchem.2010.09.065
Misra HP, Fridovich I. The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem. 1972;247(10):3170-3175. doi:10.1016/S0021-9258(19)45228-9
Aebi H. Catalase in vitro. Methods Enzymol. 1984;105:121-126. doi:10.1016/s0076-6879(84)05016-3
Beutler E, Duron O, Kelly BM. Improved method for the determination of blood glutathione. J Lab Clin Med. 1963;61:882-888. PMID: 13967893.
Da Cruz G. Use of bathocuproine for the evaluation of the antioxidant power in liquids and solutions. 2003;US patent 6613577.
Chen LY, Mehta P, Mehta JL. Oxidized LDL decreases L-arginine uptake and nitric oxide synthase protein expression in human platelets: relevance of the effect of oxidized LDL on platelet function. Circ. 1996;93(9):1740-1746. doi:10.1161/01.CIR.93.9.1740
Olas B, Nowak P, Kolodziejczyk J, Ponczek M, Wachowicz B. Protective effects of resveratrol against oxidative/nitrative modifications of plasma proteins and lipids exposed to peroxynitrite. J Nutr Biochem. 2006;17(2):96-102. doi:10.1016/j.jnutbio.2005.05.010
Witko-Sarsat V, Friedlander M, Capeillère-Blandin C, et al. Advanced oxidation protein products as a novel marker of oxidative stress in uremia. Kidney Int. 1996;49(5):1304-1313. doi:10.1038/ki.1996.186
Habeeb AFSA. Reaction of protein sulfhydryl groups with Ellman's reagent. Methods Enzymol. 1972;25(25):457-464. doi:10.1016/S0076-6879(72)25041-8
Born GVR, Cross MOJ. The aggregation of blood platelets. J Physiol. 1963;168(1):178. doi:10.1113/jphysiol.1963.sp007185
Tamang HK, Stringham EN, Tourdot BE. Platelet functional testing via high-throughput microtiter plate-based assays. Curr Protoc. 2023;3(2):668. doi:10.1002/cpz1.668
Wachowicz B, Olas B, Zbikowska HM, Buczyński A. Generation of reactive oxygen species in blood platelets. Platelets. 2002;13(3):175-182. doi:10.1080/09533710022149395
Shiri R, Yari F, Ahmadinejad M, Vaeli S, Tabatabaei MR. The caspase-3 inhibitor (peptide Z-DEVD-FMK) affects the survival and function of platelets in platelet concentrate during storage. Blood Res. 2014;49(1):49. doi:10.5045/br.2014.49.1.49
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193(1):265-275. doi:10.1016/S0021-9258(19)52451-6
Amorini AM, Tuttobene M, Lazzarino G, Denti G. Evaluation of biochemical parameters in platelet concentrates stored in glucose solution. Blood Transfus. 2007;5(1):24. doi:10.2450/2007.0019-06
Basak A. Development of a rapid and inexpensive plasma glucose estimation by two-point kinetic method based on glucose oxidase-peroxidase enzymes. Indian J Clin Biochem. 2007;22(1):156-160. doi:10.1007/BF02912902
Buhl SN, Jackson KY. Optimal conditions and comparison of lactate dehydrogenase catalysis of the lactate-to-pyruvate and pyruvate-to-lactate reactions in human serum at 25, 30, and 37 degrees C. Clin Chem. 1978;24(5):828-831. doi:10.1093/clinchem/24.5.828
Soumya R, Vani R. Vitamin C as a modulator of oxidative stress in erythrocytes of stored blood. Acta Haematol Pol. 2017;48(4):350-356. doi:10.1016/j.achaem.2017.08.005
Pallavi M, Rajashekaraiah V. Vitamin C with N-acetylcysteine ameliorates the antioxidant defenses in erythrocytes during storage. J Exp Clin Med. 2024;41(2):302-310. doi:10.52142/omujecm.41.2.14
Puskas F, Gergely Jr P, Banki K, Perl A. Stimulation of the pentose phosphate pathway and glutathione levels by dehydroascorbate, the oxidized form of vitamin C. FASEB J. 2000;14(10):1352-1361. doi:10.1096/fasebj.14.10.1352
Guo L, Qian C, Gao C, et al. Activation of the pentose phosphate pathway mitigates platelet storage lesions and improves platelet preservation quality. Thromb Res. 2025;252:109377. doi:10.1016/j.thromres.2025.109377
Çimen MB. Free radical metabolism in human erythrocytes. Clin Chim Acta. 2008;390(1-2):1-1. doi:10.1016/j.cca.2007.12.025
Ou H, Huang Z, Mo Z, Xiao J. The characteristics and roles of advanced oxidation protein products in atherosclerosis. Cardiovasc Toxicol. 2017;17(1):1-12. doi:10.1007/s12012-016-9377-8
Masselli E, Pozzi G, Vaccarezza M, et al. ROS in platelet biology: functional aspects and methodological insights. Int J Mol Sci. 2020;21(14):4866. doi:10.3390/ijms21144866
Dionisio LM, Zheng Y, Cancelas JA. Redox Control in Platelet Activity and Therapy. Antioxidants. 2025;14(11):1286. doi:10.3390/antiox14111286
Freedman JE. Oxidative stress and platelets. Arterioscler Thromb Vasc Biol. 2008;28(3):11-16. doi:10.1161/ATVBAHA.107.15917
Gulliksson H. Defining the optimal storage conditions for the long-term storage of platelets. Transfus Med Rev. 2003;17(3):209-215. doi:10.1016/S0887-7963(03)00020-8
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2026 European Journal of Clinical and Experimental Medicine
This work is licensed under a Creative Commons Attribution 4.0 International License.
Our open access policy is in accordance with the Budapest Open Access Initiative (BOAI) definition: this means that articles have free availability on the public Internet, permitting any users to read, download, copy, distribute, print, search, or link to the full texts of these articles, crawl them for indexing, pass them as data to software, or use them for any other lawful purpose, without financial, legal, or technical barriers other than those inseparable from having access to the Internet itself.
All articles are published with free open access under the CC-BY Creative Commons attribution license (the current version is CC-BY, version 4.0). If you submit your paper for publication by the Eur J Clin Exp Med, you agree to have the CC-BY license applied to your work. Under this Open Access license, you, as the author, agree that anyone may download and read the paper for free. In addition, the article may be reused and quoted provided that the original published version is cited. This facilitates freedom in re-use and also ensures that Eur J Clin Exp Med content can be mined without barriers for the research needs.
