Evaluation of platelet mitochondria integrity after treatment with Mirasol pathogen reduction technology

Abstract

Previous studies showed that Mirasol (Navigant Biotechnologies, Inc.) pathogen reduction technology (PRT) treatment resulted in an increase in platelet (PLT) glucose consumption and lactate production rates and decrease in pH in media during PLT storage. Increased glycolytic flux could result from damage to mitochondria and/or increased ATP consumption. PLT concentrates were collected by standard automated blood component collection system (Trima, Gambro BCT) procedure on Day 0 and treated with Mirasol PRT treatment on Day 1. PLT mitochondrial transmembrane potential was evaluated by staining PLTs with JC-1 followed by flow cytometry analysis. Mitochondrial enzymatic activity was measured by the MTT assay. ATP content and pH were also quantified. The values for these measurements were compared among control, untreated, and pathogen reduction technology (PRT)-treated PLTs during PLT storage for up to 7 days. No significant changes were found in pH, JC-1 signal, MTT activity, and ATP content of the PLTs immediately after PRT treatment. The treated PLTs exhibited a moderate but significantly accelerated decrease in pH and lower ATP content after 7-day storage when compared to control PLTs. Neither the JC-1 assay nor the MTT assay, however, showed a significant difference between control and treated PLTs during PLT storage. There is no evidence from these studies that Mirasol PRT treatment alters PLT mitochondrial structural and functional integrity immediately after treatment and during PLT storage. An increased demand for ATP may be the driving force for observed increases in both the glycolytic flux and the oxidative metabolism observed in treated PLTs.