Research Paper

Journal of Nanoparticle Research

, 16:2167

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

  • Anoop K. PalAffiliated withBiomedical Engineering and Biotechnology Program, University of Massachusetts Lowell Email author 
  • , Shu-Feng HsiehAffiliated withDepartment of Clinical Laboratory and Nutritional Sciences, University of Massachusetts Lowell
  • , Madhu KhatriAffiliated withBiomedical Engineering and Biotechnology Program, University of Massachusetts Lowell
  • , Jacqueline A. IsaacsAffiliated withDepartment of Mechanical, Industrial, and Manufacturing Engineering, Northeastern University
  • , Philip DemokritouAffiliated withDepartment of Environmental Health, Harvard School of Public Health
  • , Peter GainesAffiliated withDepartment of Biological Sciences, University of Massachusetts LowellCenter for High-rate Nanomanufacturing, University of Massachusetts Lowell
  • , Daniel F. SchmidtAffiliated withDepartment of Plastics Engineering, University of Massachusetts LowellCenter for High-rate Nanomanufacturing, University of Massachusetts Lowell
  • , Eugene J. RogersAffiliated withDepartment of Clinical Laboratory and Nutritional Sciences, University of Massachusetts LowellCenter for High-rate Nanomanufacturing, University of Massachusetts Lowell
  • , Dhimiter BelloAffiliated withBiomedical Engineering and Biotechnology Program, University of Massachusetts LowellDepartment of Work Environment, University of Massachusetts LowellCenter for High-rate Nanomanufacturing, University of Massachusetts Lowell Email author 

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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.

Keywords

Oxidative stress Engineered nanomaterials ROS Glutathione DCFH FRAS ESR Nanotechnology Environmental and health effects