In vitro study of platelet function confirms the contribution of the ultraviolet B (UVB) radiation in the lesions observed in riboflavin/UVB-treated platelet concentrates

Transfusion. 2015 Sep;55(9):2219-30. doi: 10.1111/trf.13123. Epub 2015 Apr 15.

Abstract

Background: Platelet inactivation technologies (PITs) have been shown to increase platelet storage lesions (PSLs). This study investigates amotosalen/ultraviolet (UV)A- and riboflavin/UVB-induced platelet (PLT) lesions in vitro. Particular attention is given to the effect of UVB alone on PLTs.

Study design and methods: Buffy coat-derived PLT concentrates (PCs) were treated with amotosalen/UVA, riboflavin/UVB, or UVB alone and compared to untreated PCs throughout storage. In vitro PLT function was assessed by blood gas and metabolite analyses, flow cytometry-based assays (CD62P, JC-1, annexin V, PAC-1), hypotonic shock response, and static adhesion to fibrinogen-coated wells.

Results: In our experimental conditions, riboflavin/UVB-treated PCs showed the most pronounced differences compared to untreated and amotosalen/UVA-treated PCs. The riboflavin/UVB treatment led to a significant increase of anaerobic glycolysis rate despite functional mitochondria, a significant increase of CD62P on Day 2, and a decrease of JC-1 aggregates and increase of annexin V on Day 7. The expression of active GPIIbIIIa (PAC-1) and the adhesion to fibrinogen was significantly increased from Day 2 of storage in riboflavin/UVB-treated PCs. Importantly, we showed that these lesions were caused by the UVB radiation alone, independently of the presence of riboflavin.

Conclusion: The amotosalen/UVA-treated PCs confirmed previously published results with a slight increase of PSLs compared to untreated PCs. Riboflavin/UVB-treated PCs present significant in vitro PSLs compared to untreated PCs. These lesions are caused by the UVB radiation alone and probably involve the generation of reactive oxygen species. The impact of these observations on clinical use must be investigated.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Annexin A5 / metabolism
  • Blood Buffy Coat / metabolism
  • Blood Buffy Coat / pathology
  • Blood Platelets / metabolism*
  • Blood Platelets / pathology
  • Blood Preservation*
  • Dual Specificity Phosphatase 2 / metabolism
  • Female
  • Flow Cytometry*
  • Furocoumarins / blood
  • Glycolysis* / drug effects
  • Glycolysis* / radiation effects
  • Humans
  • Male
  • Osmotic Pressure / drug effects
  • Osmotic Pressure / radiation effects
  • P-Selectin / metabolism
  • Platelet Function Tests
  • Riboflavin / pharmacology*
  • Time Factors
  • Ultraviolet Rays*

Substances

  • Annexin A5
  • Furocoumarins
  • P-Selectin
  • SELP protein, human
  • DUSP2 protein, human
  • Dual Specificity Phosphatase 2
  • amotosalen
  • Riboflavin