Dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay: a quantitative method for oxidative stress assessment of nanoparticle-treated cells

Autor: J.V. Castell, Guillermo Quintás, E. Burello, Laia Tolosa, A. Aranda, L. Sequedo, L. Gombau
Rok vydání: 2012
Předmět:
Male
RAPID - Risk Analysis for Products in Development
02 engineering and technology
Toxicology
medicine.disease_cause
Ferric Compounds
Polyethylene Glycols
Rats
Sprague-Dawley
chemistry.chemical_compound
Life
Polylactic Acid-Polyglycolic Acid Copolymer
Fluorescein
Cells
Cultured
Titanium
0303 health sciences
General Medicine
DCFH-DA
021001 nanoscience & nanotechnology
Fluoresceins
Silicon Dioxide
Fluorescence
Biochemistry
In vitro assays
Biological Assay
0210 nano-technology
Healthy Living
Cell Survival
Rhodamine
03 medical and health sciences
medicine
Food and Nutrition
Animals
Lactic Acid
030304 developmental biology
Nutrition
Fluorescent Dyes
Quenching (fluorescence)
Ethylene oxide
Ferrosoferric Oxide
Rats
Oleic acid
Oxidative Stress
chemistry
Titanium dioxide
Hepatocytes
Nanoparticles
ELSS - Earth
Life and Social Sciences
Reactive Oxygen Species
Oxidative stress
Polyglycolic Acid
Nuclear chemistry
Zdroj: Toxicology in Vitro; Vol 27
TOXICOLOGY IN VITRO
r-IIS La Fe. Repositorio Institucional de Producción Científica del Instituto de Investigación Sanitaria La Fe
instname
Toxicology in Vitro, 2, 27, 954–963
Toxicology in Vitro
ISSN: 1879-3177
0887-2333
Popis: No consensus exists on how to address possible toxicity of nanomaterials as they interfere with most in vitro screening tests based on colorimetric and fluorimetric probes such as the dichloro-dihydrofluorescein diacetate (DCFH-DA) assay for detection of oxidative species. In the present research, nanomaterial interaction with DCFH-DA was studied in relation to its nature and/or assay conditions (cell-based and time exposure) by incubating Rhodamine (Rhd)-labeled 25 nm and 50 nm silica (SiO2), naked and oleic acid coated magnetite, (Fe3O4) and maghemite (Fe2O3) iron oxide, titanium dioxide (TiO2) and poly(ethylene oxide)-poly(lactide/glycolide) acid (PLGA-PEO) nanoparticles (NPs) with metabolically active rat hepatocytes for 4 and 24-h periods. Data indicated that nanoparticle uptake correlated with quenching of dye fluorescence emission. In spite of their masking effect, the oxidative potential of NPs could be detected at a limited threshold concentration when exposed for periods of time longer than those frequently used for this test. However, changes in the experimental conditions did not systematically result in free radical formation for all nanomaterials tested. Overall data indicate that despite the quenching effect of nanoparticles on DCFH-DA assay, it can be considered as a useful tool for quantitative measurement of NPs-induced oxidative stress by minor modifications of standardized protocols.
Databáze: OpenAIRE
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