Figure 5

Nanomaterial dependent ROS generation and lysosomal membrane destabilization (LMD) in 3T3, hT, and RAW cells. (a) ROS generation was quantified using 2',7'-dichlorodihydrofluorescein diacetate (H₂DCFDA) for untreated cells and compared to cells exposed with each test nanomaterial for 2 hours at 100 μg/ml. LMD was visualized via acridine orange staining in (b) 3T3, (c) hT, and (d) RAW cells exposed to nanomaterials at 100 μg/ml for 4 hrs. In control (healthy) cells, lysosomes can be seen as red-orange granules and cytoplasm has a diffuse green fluorescence. In cells with lysosomal membrane damage, lysosomes exhibit a shift from red-orange to a green color and overall intensity of green fluorescence is increased in these cells. (e) Quantification of LMD after 4 hours of exposure to nanomaterials at 100 μg/ml was done via acridine orange relocation technique. For each nanomaterial, bracket with a (*) indicates significant difference between different cell types by ANOVA. (*) above nanomaterial columns indicates significant difference from untreated cells by Bonferroni test.