Advertisement

Journal of Thrombosis and Thrombolysis

, Volume 48, Issue 3, pp 430–438 | Cite as

Platelet spreading on fibrinogen matrix, a reliable and sensitive marker of platelet functional activity during storage

  • Ehteramolsadat Hosseini
  • Mehran GhasemzadehEmail author
  • Elaheh Azizvakili
  • Pezhman Beshkar
Article

Abstract

Upon platelet activation, inside-out signals synergistically induced by a variety of agonists and adhesion molecules can enhance the affinity of platelet main integrin, αIIbβ3 to its ligands. Integrin ligation with fibrinogen induces potent signals which develop platelet function including aggregation, release and spreading of which platelet spreading is considered as a major early consequence of αIIbβ3 outside-in signaling. Study presented here evaluated platelet spreading on fibrinogen matrix as a marker of platelet activation during storage. PRP-platelet concentrates were subjected to flowcytometry analysis and the expression levels of P-selectin, CD61, GPIbα and active conformation of the αIIbβ3 (PAC-1 binding) were examined on day 0, 1, 3 and 5 post-storage. Concurrently platelet adhesion and spreading on fibrinogen matrix, glucose concentration and LDH activity were also determined at the same intervals. Results showed significant decreases in platelet spreading on fibrinogen matrix during storage. Spreading was dominant pattern of adhesion till the first day of storage. In 3 day-stored platelets, filopodial or lamellipodial formation was dominant event whereas 5 day-stored platelets simply adhered to fibrinogen with less protrusion formation and partially failed to spread. Compared to simple adhesion, reduction of platelet spreading was also more significantly correlated with the usual markers of platelet storage lesion including P-selectin (r = − 0.88; p < 0.0001) and GPIbα expression (r = 0.76; p = 0.0001), PAC-1 binding (r = 0.66; p = 0.001), glucose concentration and LDH activity. Taken together, we introduced platelet spreading on fibrinogen matrix as a reliable and sensitive marker of platelets functional activity during storage. As a valid marker which is directly and obviously relevant to the platelet functional capacities, the rapid reduction of platelet spreading during storage overshadows other markers of platelet storage lesion while raising serious question about the quality of 5 day-stored platelets.

Keywords

Adhesion Fibrinogen Integrin alpha IIb beta 3 (αIIbβ3Platelet spreading Platelet storage lesion Platelet transfusion Thrombosis 

Notes

Acknowledgements

This work was part of Dr. Ghasemzadeh and Dr. Hosseini’s approved projects (Nos. 1394-01-33-1861 and 1396-06-33-2036) which were supported by Iranian blood transfusion organization and High Institute for Research and Education in Transfusion Medicine in Iran, also approved by local ethic committee. The authors declare no conflict of interests.

Author contributions

MG supervised and designed the study, did the experiments, analyzed the data and wrote the paper. EH did some experiments, helped with analyzing data and co-wrote the paper. EA did some experiments as a part of her MS project. PB did some experiments.

References

  1. 1.
    Shrivastava M (2009) The platelet storage lesion. Transfus Apher Sci 41(2):105–113.  https://doi.org/10.1016/j.transci.2009.07.002 CrossRefGoogle Scholar
  2. 2.
    Hosseini E, Ghasemzadeh M, Nassaji F, Jamaat ZP (2017) GPVI modulation during platelet activation and storage: its expression levels and ectodomain shedding compared to markers of platelet storage lesion. Platelets 28(5):498–508CrossRefGoogle Scholar
  3. 3.
    Hosseini E, Ghasemzadeh M, Atashibarg M, Haghshenas M (2019) ROS scavenger, N-acetyl-l-cysteine and NOX specific inhibitor, VAS2870 reduce platelets apoptosis while enhancing their viability during storage. Transfusion 59(4):1333–1343CrossRefGoogle Scholar
  4. 4.
    Ghasemzadeh M (2009) Investigation of signaling cross-talk between platelets and neutrophils. Monash University. Faculty of Medicine, Nursing and Health Sciences. Australian Centre for Blood DiseasesGoogle Scholar
  5. 5.
    Hosseini E, Ghasemzadeh M (2012) Different stages of platelet adhesion to the site of vascular injury. Iranian Journal of Blood and Cancer 4(3):133–142Google Scholar
  6. 6.
    Seghatchian J, Krailadsiri P (1997) The platelet storage lesion. Transfus Med Rev 11(2):130–144CrossRefGoogle Scholar
  7. 7.
    Mehrpoori M, Hosseini E, Amini Kafi-Abad S, Ghasemzadeh M (2015) The effect of pre-storage leukoreduction on the levels of expression and shedding of the pro-inflammatory molecule P-Sel in random PRP platelets. Sci J Iran Blood Transfus Org 12(2):153–162Google Scholar
  8. 8.
    Sharifrazi M, GhasemzadehM SKE, Hosseini E (2015) The effect of pre-storage leukoreduction on the levels of expression of the pro-inflammatory molecule CD40 ligand in random PRP platelets. Razi J Med Sci 22(141):38–46 (in persian) Google Scholar
  9. 9.
    Bontekoe IJ, van der Meer PF, van den Hurk K, Verhoeven AJ, de Korte D (2017) Platelet storage performance is consistent by donor: a pilot study comparing “good” and “poor” storing platelets. Transfusion 57(10):2373–2380CrossRefGoogle Scholar
  10. 10.
    Hosseini E, Beshkar P, Ghasemzadeh M (2018) Reverse correlations of collagen-dependent platelet aggregation and adhesion with GPVI shedding during storage. J Thromb Thrombolysis 46(4):534–540CrossRefGoogle Scholar
  11. 11.
    George J, Pickett E, Heinz R (1988) Platelet membrane glycoprotein changes during the preparation and storage of platelet concentrates. Transfusion 28(2):123–126CrossRefGoogle Scholar
  12. 12.
    Metcalfe P, Williamson LM, Reutelingsperger CP, Swann I, Ouwehand WH, Goodall AH (1997) Activation during preparation of therapeutic platelets affects deterioration during storage: a comparative flow cytometric study of different production methods. Br J Haematol 98(1):86–95CrossRefGoogle Scholar
  13. 13.
    Albanyan AM, Harrison P, Murphy MF (2009) Markers of platelet activation and apoptosis during storage of apheresis- and buffy coat-derived platelet concentrates for 7 days. Transfusion 49(1):108–117.  https://doi.org/10.1111/j.1537-2995.2008.01942.x CrossRefGoogle Scholar
  14. 14.
    Gardiner EE, Al-Tamimi M, Andrews RK, Berndt MC (2012) Platelet receptor shedding. Platelets and megakaryocytes, vol 3. Additional protocols and perspectives. Springer, New York, pp 321–339CrossRefGoogle Scholar
  15. 15.
    Andrews RK, Karunakaran D, Gardiner EE, Berndt MC (2007) Platelet receptor proteolysis. Arterioscler Thromb Vasc Biol 27(7):1511–1520CrossRefGoogle Scholar
  16. 16.
    Wood B, Padula MP, Marks DC, Johnson L (2016) Refrigerated storage of platelets initiates changes in platelet surface marker expression and localization of intracellular proteins. Transfusion 56(10):2548–2559CrossRefGoogle Scholar
  17. 17.
    Sandgren P, Diedrich B (2015) Pathogen inactivation of double-dose buffy-coat platelet concentrates photochemically treated with amotosalen and UVA light: preservation of in vitro function. Vox Sang 108(4):340–349CrossRefGoogle Scholar
  18. 18.
    Johnson L, Schubert P, Tan S, Devine DV, Marks DC (2016) Extended storage and glucose exhaustion are associated with apoptotic changes in platelets stored in additive solution. Transfusion 56(2):360–368CrossRefGoogle Scholar
  19. 19.
    Beshkar P, Hosseini E, Ghasemzadeh M (2018) Superior integrin activating capacity and higher adhesion to fibrinogen matrix in buffy coat-derived platelet concentrates (PCs) compared to PRP-PCs. Transfus Apher Sci 57(1):76–81.  https://doi.org/10.1016/j.transci.2017.12.003 CrossRefGoogle Scholar
  20. 20.
    Ghasemzadeh M, Hosseini E, Roudsari ZO, Zadkhak P (2018) Intraplatelet reactive oxygen species (ROS) correlate with the shedding of adhesive receptors, microvesiculation and platelet adhesion to collagen during storage: does endogenous ROS generation downregulate platelet adhesive function? Thromb Res 163:153–161CrossRefGoogle Scholar
  21. 21.
    Li Z, Delaney MK, O’brien KA, Du X (2010) Signaling during platelet adhesion and activation. Arterioscler Thromb Vasc Biol 30(12):2341–2349CrossRefGoogle Scholar
  22. 22.
    Shattil SJ, Newman PJ (2004) Integrins: dynamic scaffolds for adhesion and signaling in platelets. Blood 104(6):1606–1615CrossRefGoogle Scholar
  23. 23.
    Nieswandt B, Bergmeier W, Eckly A, Schulte V, Ohlmann P, Cazenave J-P, Zirngibl H, Offermanns S, Gachet C (2001) Evidence for cross-talk between glycoprotein VI and Gi-coupled receptors during collagen-induced platelet aggregation. Blood 97(12):3829–3835CrossRefGoogle Scholar
  24. 24.
    Nieswandt B, Schulte V, Zywietz A, Gratacap M-P, Offermanns S (2002) Costimulation of Gi-and G12/G13-mediated signaling pathways induces integrin αIIbβ3 activation in platelets. J Biol Chem 277(42):39493–39498CrossRefGoogle Scholar
  25. 25.
    Kulkarni S, Woollard KJ, Thomas S, Oxley D, Jackson SP (2007) Conversion of platelets from a proaggregatory to a proinflammatory adhesive phenotype: role of PAF in spatially regulating neutrophil adhesion and spreading. Blood 110(6):1879–1886CrossRefGoogle Scholar
  26. 26.
    Vanderlinde RE (1985) Measurement of total lactate dehydrogenase activity. Ann Clin Lab Sci 15(1):13–31Google Scholar
  27. 27.
    Di Minno G, Capitanio AM, Thiagarajan P, Martinez J, Murphy S (1983) Exposure of fibrinogen receptors on fresh and stored platelets by ADP and epinephrine as single agents and as a pair. Blood 61(6):1054–1059Google Scholar
  28. 28.
    Goodall A (1991) Platelet activation during preparation and storage of concentrates: detection by flow cytometry. Blood Coagul Fibrinol 2(2):377–382CrossRefGoogle Scholar
  29. 29.
    Lozano M, Estebanell E, Cid J, Diaz-Ricart M, Mazzara R, Ordinas A, Escolar G (1999) Platelet concentrates prepared and stored under currently optimal conditions: minor impact on platelet adhesive and cohesive functions after storage. Transfusion 39(9):951–959CrossRefGoogle Scholar
  30. 30.
    Tynngård N, Wallstedt M, Södergren AL, Faxälv L, Ramström S (2015) Platelet adhesion changes during storage studied with a novel method using flow cytometry and protein-coated beads. Platelets 26(2):177–185CrossRefGoogle Scholar
  31. 31.
    Janes S, Wilson D, Chronos N, Goodall A (1993) Evaluation of whole blood flow cytometric detection of platelet bound fibrinogen on normal subjects and patients with activated platelets. Thromb Haemost 70(4):659–666Google Scholar
  32. 32.
    Sandgren P, Saeed K (2011) Storage of buffy-coat-derived platelets in additive solution: in vitro effects on platelets of the air bubbles and foam included in the final unit. Blood Transfus 9(2):182Google Scholar
  33. 33.
    Johnson L, Hyland R, Tan S, Tolksdorf F, Sumian C, Seltsam A, Marks D (2016) In vitro quality of platelets with low plasma carryover treated with ultraviolet C light for pathogen inactivation. Transfus Med Hemother 43(3):190–197CrossRefGoogle Scholar
  34. 34.
    Kulkarni S, Jackson SP (2004) Platelet factor XIII and calpain negatively regulate integrin αIIbβ3 adhesive function and thrombus growth. J Biol Chem 279(29):30697–30706CrossRefGoogle Scholar
  35. 35.
    Mehrpoori M, Hosseini E, Amini Kafi-Abad S (2015) The effect of prestorage leukoreduction on the levels of expression and shedding of the pro-inflammatory molecule P-Sel in random PRP platelets. Sci J Iran Blood Transfus Organ 12(2):153–162Google Scholar
  36. 36.
    Fu Q, Garnham CP, Elliott ST, Bovenkamp DE, Van Eyk JE (2005) A robust, streamlined, and reproducible method for proteomic analysis of serum by delipidation, albumin and IgG depletion, and two-dimensional gel electrophoresis. Proteomics 5(10):2656–2664.  https://doi.org/10.1002/pmic.200402048 CrossRefGoogle Scholar
  37. 37.
    Furman MI, Krueger LA, Linden MD, Barnard MR, Frelinger AL, Michelson AD (2004) Release of soluble CD40L from platelets is regulated by glycoprotein IIb/IIIa and actin polymerization. J Am Coll Cardiol 43(12):2319–2325CrossRefGoogle Scholar
  38. 38.
    Garcia BA, Smalley DM, Cho H, Shabanowitz J, Ley K, Hunt DF (2005) The platelet microparticle proteome. J Proteome Res 4(5):1516–1521.  https://doi.org/10.1021/pr0500760 CrossRefGoogle Scholar
  39. 39.
    Levin E, Culibrk B, Gyöngyössy-Issa MI, Weiss S, Scammell K, LeFresne W, Jenkins C, Devine DV (2008) Implementation of buffy coat platelet component production: comparison to platelet-rich plasma platelet production. Transfusion 48(11):2331–2337CrossRefGoogle Scholar
  40. 40.
    Ohto H, Nollet KE (2011) Overview on platelet preservation: better controls over storage lesion. Transfus Aphere Sci 44(3):321–325CrossRefGoogle Scholar
  41. 41.
    Vetlesen A, Mirlashari MR, Ezligini F, Kjeldsen-Kragh J (2007) Evaluation of platelet activation and cytokine release during storage of platelet concentrates processed from buffy coats either manually or by the automated OrbiSac system. Transfusion 47(1):126–132CrossRefGoogle Scholar
  42. 42.
    Tynngård N, Studer M, Lindahl TL, Trinks M, Berlin G (2008) The effect of gamma irradiation on the quality of apheresis platelets during storage for 7 days. Transfusion 48(8):1669–1675CrossRefGoogle Scholar
  43. 43.
    Johnson L, Schubert P, Tan S, Devine DV, Marks DC (2015) Extended storage and glucose exhaustion are associated with apoptotic changes in platelets stored in additive solution. Transfusion 56(2):360–368CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Ehteramolsadat Hosseini
    • 1
  • Mehran Ghasemzadeh
    • 1
    • 2
    • 4
    Email author
  • Elaheh Azizvakili
    • 1
  • Pezhman Beshkar
    • 1
    • 3
  1. 1.Blood Transfusion Research CenterHigh Institute for Research and Education in Transfusion MedicineTehranIran
  2. 2.Australian Centre for Blood DiseasesMonash UniversityMelbourneAustralia
  3. 3.Cellular and Molecular Research Centre, Basic Health Science InstituteShahrekord University of Medical ScienceSharekordIran
  4. 4.Blood Transfusion Research CentreHigh Institute for Research and Education in Transfusion MedicineTehranIran

Personalised recommendations