Image- and Fluorescence-Based Test Shows Oxidant-Dependent Damages in Red Blood Cells and Enables Screening of Potential Protective Molecules.

Details

Serval ID
serval:BIB_5D01C543DB17
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Image- and Fluorescence-Based Test Shows Oxidant-Dependent Damages in Red Blood Cells and Enables Screening of Potential Protective Molecules.
Journal
International journal of molecular sciences
Author(s)
Bardyn M., Allard J., Crettaz D., Rappaz B., Turcatti G., Tissot J.D., Prudent M.
ISSN
1422-0067 (Electronic)
ISSN-L
1422-0067
Publication state
Published
Issued date
20/04/2021
Peer-reviewed
Oui
Volume
22
Number
8
Pages
4293
Language
english
Notes
Publication types: Journal Article
Publication Status: epublish
Abstract
An increase of oxygen saturation within blood bags and metabolic dysregulation occur during storage of red blood cells (RBCs). It leads to the gradual exhaustion of RBC antioxidant protective system and, consequently, to a deleterious state of oxidative stress that plays a major role in the apparition of the so-called storage lesions. The present study describes the use of a test (called TSOX) based on fluorescence and label-free morphology readouts to simply and quickly evaluate the oxidant and antioxidant properties of various compounds in controlled conditions. Here, TSOX was applied to RBCs treated with four antioxidants (ascorbic acid, uric acid, trolox and resveratrol) and three oxidants (AAPH, diamide and H <sub>2</sub> O <sub>2</sub> ) at different concentrations. Two complementary readouts were chosen: first, where ROS generation was quantified using DCFH-DA fluorescent probe, and second, based on digital holographic microscopy that measures morphology alterations. All oxidants produced an increase of fluorescence, whereas H <sub>2</sub> O <sub>2</sub> did not visibly impact the RBC morphology. Significant protection was observed in three out of four of the added molecules. Of note, resveratrol induced diamond-shape "Tirocytes". The assay design was selected to be flexible, as well as compatible with high-throughput screening. In future experiments, the TSOX will serve to screen chemical libraries and probe molecules that could be added to the additive solution for RBCs storage.
Keywords
Physical and Theoretical Chemistry, Inorganic Chemistry, Organic Chemistry, Spectroscopy, Molecular Biology, Catalysis, General Medicine, Computer Science Applications, antioxidant, digital holographic microscopy, fluorescence, high-throughput screening, oxidant, red blood cell, transfusion
Pubmed
Web of science
Open Access
Yes
Create date
22/04/2021 7:38
Last modification date
08/05/2021 6:32
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