NDLI: Screening for oxidative damage by engineered nanomaterials: a comparative evaluation of FRAS and DCFH

Content Provider	Springer Nature Link
Author	Pal, Aop K. Hsieh, Shu Feng Khatri, Madhu Isaacs, Jacqueline A. Demokritou, Philip Gaines, Peter Schmidt, Daniel F. Rogers, Eugene J. Bello, Dhimiter
Copyright Year	2013
Abstract	Several acellular assays are routinely used to measure oxidative stress elicited by engineered nanomaterials (ENMs), yet little comparative evaluations of such methods exist. This study compares for the first time the performance of the dichlorofluorescein (DCFH) assay which measures reactive oxygen species (ROS) generation, to that of the ferric-reducing ability of serum (FRAS) assay, which measures biological oxidant damage in serum. A diverse set of 28 commercially important and extensively characterized ENMs were tested on both the assays. Intracellular oxidative stress was also assessed on a representative subset of seven ENMs in THP-1 (phorbol 12-myristate 13-acetate matured human monocytes) cells. Associations between assay responses and ENM physicochemical properties were assessed via correlation and regression analysis. DCFH correlated strongly with FRAS after dose normalization for mass (R $^{2}$ = 0.78) and surface area (R $^{2}$ = 0.68). Only 10/28 ENMs were positive in DCFH versus 21/28 in FRAS. Both assays were strongly associated with specific surface area and transition metal content. Qualitatively, a similar response ranking was observed for acellular FRAS and intracellular reduced:oxidized glutathione ratio (GSH:GSSG) in cells. Quantitatively, weak correlation was found between intracellular GSSG and FRAS or DCFH (R $^{2}$ < 0.25) even after calculating effective dose to cells. The FRAS assay was more sensitive than DCFH, especially for ENMs with low to moderate oxidative damage potential, and may serve as a more biologically relevant substitute for acellular ROS measurements of ENMs. Further in vitro and in vivo validations of FRAS against other toxicological endpoints with larger datasets are recommended.
Starting Page	1
Ending Page	20
Page Count	20
File Format	PDF
ISSN	13880764
Journal	Journal of Nanoparticle Research
Volume Number	16
Issue Number	2
e-ISSN	1572896X
Language	English
Publisher	Springer Netherlands
Publisher Date	2014-01-03
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Oxidative stress Engineered nanomaterials ROS Glutathione DCFH FRAS ESR Nanotechnology Environmental and health effects Inorganic Chemistry Characterization and Evaluation of Materials Physical Chemistry Optics, Optoelectronics, Plasmonics and Optical Devices
Content Type	Text
Resource Type	Article
Subject	Chemistry Nanoscience and Nanotechnology Atomic and Molecular Physics, and Optics Condensed Matter Physics Bioengineering Modeling and Simulation

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