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Atomic, Molecular and Chemical Physics, Biochemical research methods, Nano-materials
The need for rapid and cost effective pre-screening protocols of the toxicological response of the vast array of emerging nanoparticle types is apparent and the emerging consensus on the paradigm of oxidative stress by generation of intracellular reactive oxygen species as a primary source of the toxic response suggests the development of acellular assays to screen for nanoparticle surface reactivity. This study explores the potential of the monoamine oxidase A (MAO-A) enzyme based assay with polymeric dendrimers as cofactors and serotonin as substrate, which generates H2O2, quantified by the conversion of the Carboxy-H2DCFDA dye to its fluorescent form. A range of generations of both PAMAM (poly(amido amine)) (G4-G7) and PPI (poly(propylene imine)) (G0-G4) dendritic polymer nanoparticles are used as test particles to validate the quantitative nature of the assay response as a function of nanoparticle physico-chemical properties. The assay is well behaved as a function of dose, over low dose ranges and the acellular reaction rate (ARR) is well correlated with the number of surface amino groups for the combined dendrimer series. For each series, the ARR is also well correlated with the previously documented cytotoxicity, although the correlation is substantially different for each series of dendrimers, pointing to the additional importance of cellular uptake rates in the determination of toxicity.
Byrne, H.J., Maher, M. A., & Khalid, H. (2016) Acellular reactivity of polymeric dendrimer nanoparticles as an indicator of oxidative stress in vitro”, Analytical and Bioanalytical Chemistry, 408, 695-703 (2016). doi:10.1007/s00216-015-9171-7