Reactive oxygen species (ROS) induced cytokine production and cytotoxicity of PAMAM dendrimers in J774A.1 cells

• Published in: Toxicology and applied pharmacology 2010-07, Vol.246 (1), p.91-99
• Main Authors: Naha, Pratap C., Davoren, Maria, Lyng, Fiona M., Byrne, Hugh J.
• Format: Article
• Language: English
• Subjects: 60 APPLIED LIFE SCIENCES; Amino groups; ANIMAL CELLS; ANIMALS; APOPTOSIS; BIOASSAY; Biological and medical sciences; Cell Line, Tumor; Chemokine CXCL2 - analysis; Chemokine CXCL2 - biosynthesis; CONNECTIVE TISSUE CELLS; Cytokine production; Cytokines - analysis; Cytokines - biosynthesis; Cytotoxicity Tests, Immunologic; Dendrimers - toxicity; ELEMENTS; ENZYME IMMUNOASSAY; Female; GROWTH FACTORS; IMMUNOASSAY; IN VITRO; Interleukin-6 - analysis; Interleukin-6 - biosynthesis; LYMPHOKINES; Macrophage; MACROPHAGES; Macrophages - chemistry; Macrophages - drug effects; Macrophages - metabolism; MAMMALS; Medical sciences; MICE; Mice, Inbred BALB C; MITOGENS; Nanotoxicology; NONMETALS; ORGANIC COMPOUNDS; Oxazines; OXYGEN; PAMAM dendrimers; PHAGOCYTES; PROTEINS; RADIATION, THERMAL, AND OTHER ENVIRONMENTAL POLLUTANT EFFECTS ON LIVING ORGANISMS AND BIOLOGICAL MATERIALS; Reactive oxygen species; Reactive Oxygen Species - analysis; RODENTS; SOMATIC CELLS; TOXICITY; Toxicology; Tumor Necrosis Factor-alpha - analysis; Tumor Necrosis Factor-alpha - biosynthesis; VERTEBRATES; Xanthenes
• ISSN: 0041-008X; 1096-0333
• DOI: 10.1016/j.taap.2010.04.014

The immunotoxicity of three generations of polyamidoamine (PAMAM) dendrimers (G-4, G-5 and G-6) was evaluated in mouse macrophage cells in vitro. Using the Alamar blue and MTT assays, a generation dependent cytotoxicity of the PAMAM dendrimers was found whereby G-6 > G-5 > G-4. The toxic response of the PAMAM dendrimers correlated well with the number of surface primary amino groups, with increasing number resulting in an increase in toxic response. An assessment of intracellular ROS generation by the PAMAM dendrimers was performed by measuring the increased fluorescence as a result of intracellular oxidation of Carboxy H 2DCFDA to DCF both quantitatively using plate reader and qualitatively by confocal laser scanning microscopy. The inflammatory mediators macrophage inflammatory protein-2 (MIP-2), tumour necrosis factor-α (TNF-α) and interleukin-6, (IL-6) were measured by the enzyme linked immunosorbant assay (ELISA) following exposure of mouse macrophage cells to PAMAM dendrimers. A generation dependent ROS and cytokine production was found, which correlated well with the cytotoxicological response and therefore number of surface amino groups. A clear time sequence of increased ROS generation (maximum at ∼ 4 h), TNF-α and IL-6 secretion (maximum at ∼ 24 h), MIP-2 levels and cell death (∼ 72 h) was observed. The intracellular ROS generation and cytokine production induced cytotoxicity point towards the mechanistic pathway of cell death upon exposure to PAMAM dendrimers.