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Fig. 6 | Particle and Fibre Toxicology

Fig. 6

From: Macrophage-mediated tissue response evoked by subchronic inhalation of lead oxide nanoparticles is associated with the alteration of phospholipases C and cholesterol transporters

Fig. 6

Liver after 11-week PbO NP inhalation. A Ferritin agglomerate (sized 112 nm). B Hepatocyte mitochondria (mi) with agglomerates of PbO nanoparticles (arrowheads) surrounded with electron-dense matrix. C Detail of agglomerates of PbO nanoparticles (size range 10–40 nm). D Hepatocyte mitochondrion with ROI windows analyzed inside. E SEM in transmission mode, using TESCAN RSTEM detector, and energy-dispersive X-ray spectroscopy (X-EDS) of lead treated samples of liver. Data obtained from two ROI windows analyzed inside hepatocyte mitochondria (D—with and without NPs). Spectra were compared with a reference sample and analyzed using Oxford AZtec. Presence of Pb was confirmed by comparing reference Spectrum 31 (without NPs) and non-reference Spectrum 30 (with NPs, arrowhead on image D). The peak displays increased signal at a spectral position of Pb (arrow on graph E). Nickel was observed during analysis as it was issued from the grid and osmium from post-fixation. F Liver in control and PbO NP treated animals (HE staining); bv blood vessels, he hepatocytes, fn focal necrosis. Scale bar in all panels = 100 µm. G Collagen fibers (green) in liver are around blood vessel (bv) in both groups (GT staining). There is no presence of fibrosis in focal necrosis (fn). Scale bar in all panels = 100 µm. H Detection of CD68-positive cells (marker of macrophages, Kc, Kupffer cells) in liver (arrows). Scale bar in panels = 100 µm. I The number of macrophages was significantly increased compared with the control group (the graphs values indicate average ± SD; *p < 0.05). J TEM images of hepatocytes (he) with lipid droplets (arrows), Ito cell (It) with lipid droplets (arrow), and Kupffer cell (Kc) with phagosomes. Arrowhead displays cholesterol crystal in phagosome. Scale bars are displayed individually for each picture. K Gene expression of receptors CD36, SR-A1, SR-B1, Abca1, Abcg1 and phospholipases C after PbO NP inhalation. The graphs values indicate average ± SD; *p < 0.05, **p < 0.01 by unpaired t-test. L Protein expression level of receptor CD36 after PbO NP inhalation. The quantitative comparison of CD36 level was normalized to GAPDH level. M Protein expression level of phospholipase PLCβ1 after PbO NPs treatment. The quantitative comparison of PLCβ1 level was normalized to GAPDH level

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