Table 1.
Characterization of the GO, rGO and the reference P90.
Table 2.
BAL fluid cell counts (x103) (mean ±SEM) in mice at day 1, 3, 28 and 90 post exposure to VC (0.1% TW80), GO, rGO or Printex90 at doses 0, 18, 54 or 162 μg/mouse (n = 7–8).
Fig 1.
Graphical presentation of the number of neutrophils in bronchoalveolar lavage (Mean ± SEM) from at day 1, 3, 28 and 90 following exposure to VC, GO, rGO or P90 (n = 7–8).
*, ** and ***: Statistically significantly different from corresponding VC at level p < 0.05, p < 0.01, p < 0.001, respectively. #: GO statistically significantly different from corresponding rGO group at level p <0.001. For group GO 18 μg/mouse at day 1, the symbol is larger than the corresponding error bar. Therefore, the error bar is not visible (SEM is shown in Table 2).
Table 3.
mRNA expression level of Saa3 and Saa1 in lung and liver of mice 1, 3, 28 and 90 days post exposure to VC, GO, rGO or P90 at doses 0, 18, 54 or 162 μg/mouse.
Fig 2.
mRNA expression level of the acute phase response genes Saa3 in lung and Saa1 in liver 1, 3, 28 and 90 days following exposure to VC, GO, rGO and P90 (n = 7–8) at doses 0, 18, 54 and 162 μg/mouse.
Expression of mRNA was normalized to 18S rRNA and multiplied by 107. Statistical analysis for P90 compared to VC was performed separately using a t-test. *, **, ***: Statistically significantly different from corresponding VC at level p < 0.05, p < 0.01, p < 0.001, respectively.
Fig 3.
H&E stained histopathological lung sections of mice 3 days post exposure.
VC (A, B) and GO 162 μg/mouse (C, D, E, F). (A and B) No pathological changes. (C) Patchy appearance of acute pulmonary inflammation in areas with GO deposits (black arrows) in the parenchyma distal to the terminal and respiratory bronchioles, alveolar ducts, alveoli. (D) Free GO deposits (red arrow) and within alveolar cells (black arrows) in inflammatory area. Accumulation of granulocytes (green arrow). Hyperplastic type II cells (blue arrow). Congestion of vessels (CV). Alveolar granular exudate (AGE). (E) Patchy inflammation in peripheral section sites with GO deposits. Alveolar macrophage with GO in inflammatory lesion and in alveoli (black arrows), polymorphonuclear leukocytes (green arrow). Alveolar granular exudate (AGE). Hyperplastic type II cells (blue arrows). (F) Perivascular lymphoid accumulation (PVLA) with GO deposits.
Fig 4.
H&E stained histopathological lung sections of mice 90 days post exposure.
VC (A-C), GO 18 μg/mouse (D-F) or rGO 162 μg/mouse (G-I). (A-C) No pathological changes. (D-F) GO appeared as dark-brown pigments. Scattered prominent perivascular lymphoid accumulation (PVLA). Granuloma formation (GL) containing GO and macrophages with GO in alveoli (black arrows). Rare prominent perivascular lymphocytic accumulation (AGE). (G-I) Scarce accumulation of compact black rGO agglomerates (red arrows) and minimal tissue reactions.
Table 4.
Level of DNA damage (Mean ± SEM) in BAL, lung and liver assessed with the comet assay (% DNA) 1, 3, 28 and 90 days post exposure to VC, GO, rGO or P90 (n = 7–8).