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Fig. 4 | Particle and Fibre Toxicology

Fig. 4

From: Direct stimulation of human fibroblasts by nCeO2 in vitro is attenuated with an amorphous silica coating

Fig. 4

nCeO2, but not amsCeO2, directly induces the formation of fibroblastic nodules in NHLFs in a dose dependent manner. Fibroblasts were plated on poly-L-lysine coated coverslips and treated with the indicated nanoparticle at 0.006 or 0.2 μg/cm2 for 48 h. Cells were then stained for collagen I and DAPI and fibroblastic nodule formation was determined for each condition as the number of nodules formed per well. (a) Representative images of nodule formation in response to 0.2 μg/cm2 of the indicated particles. Collagen I expression is shown in green and nuclear stain (DAPI) in blue. (b) Brightfield images of the fibroblastic nodules (arrows) were used to quantitate the number of nodules formed per well in response to each of the indicated nanoparticles. Statistical significance is indicated as *p < 0.05, **p < 0.01, and ***p < 0.001 compared to non-treated (NT) conditions and ###p < 0.001 for nCeO2 compared to amsCeO2. (c) Confocal microscopy Z-stack analysis with orthogonal views (upper panels) of the X-Z plane (right) and Y-Z plane (top). Three dimensional reconstruction was generated (bottom panels) based on the results of the Z-stack images and demonstrated increased thickness of fibrotic nodule clusters formed in response to nCeO2 and TGFβ as compared to the untreated monolayer of NHLFs, indicating the mounding of the cells within the nodule

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