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Effects of the Presence or Absence of a Protein Corona on Silica Nanoparticle Uptake and Impact on Cells

Lesniak, Anna, Fenaroli, Federico, Monopoli, Marco P., Åberg, Christoffer, Dawson, Kenneth A., Salvati, Anna·2012·ACS Nano
DOI 10.1021/nn300223w
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Figure 1 A) Kinetics of uptake of 25 µg/ml fluorescently labelled 50 nm silica nanoparticles in complete medium (cMEM, empty symbols) and serum free medium (SF, filled symbols) by A549 cells, as determined by flow cytometry. Error bars are the standard deviation of the mean cell fluorescence intensity averaged over 3 replicas. B) The curve in complete medium (from panel A) alone. C) Phase contrast images of A549 cells exposed for 2 h to 25 and 100 µg/ml of 50 nm silica nanoparticles in cMEM and SF. Several cells assumed a spherical shape after exposure to silica nanoparticles in serum free medium (the effect was proportional to nanoparticle concentration). The same was not observed in cells grown in the only SF or cells exposed to the same nanoparticles in cMEM. D) ATP levels of untreated A549 cells and A549 cells exposed to different doses of 50 nm silica nanoparticles in cMEM and SF for 24 h, after normalisation for the ATP content of untreated cells in the same conditions. A
Figure 1 A) Kinetics of uptake of 25 µg/ml fluorescently labelled 50 nm silica nanoparticles in complete medium (cMEM, empty symbols) and serum free medium (SF, filled symbols) by A549 cells, as determined by flow cytometry. Error bars are the standard deviation of the mean cell fluorescence intensity averaged over 3 replicas. B) The curve in complete medium (from panel A) alone. C) Phase contrast images of A549 cells exposed for 2 h to 25 and 100 µg/ml of 50 nm silica nanoparticles in cMEM and SF. Several cells assumed a spherical shape after exposure to silica nanoparticles in serum free medium (the effect was proportional to nanoparticle concentration). The same was not observed in cells grown in the only SF or cells exposed to the same nanoparticles in cMEM. D) ATP levels of untreated A549 cells and A549 cells exposed to different doses of 50 nm silica nanoparticles in cMEM and SF for 24 h, after normalisation for the ATP content of untreated cells in the same conditions. ACC-BY-NC-SA · From the PDF
Figure 2 Transmission Electron Microscopy images of A549 cells exposed to 100 μg/ml 50 nm silica nanoparticles in complete and serum free medium for 4 h (A and C respectively) and 24 h (B and D
Figure 2 Transmission Electron Microscopy images of A549 cells exposed to 100 μg/ml 50 nm silica nanoparticles in complete and serum free medium for 4 h (A and C respectively) and 24 h (B and DCC-BY-NC-SA · From the PDF
Figure 3 Details of typical nanoparticle intracellular locations: representative images from cells
Figure 3 Details of typical nanoparticle intracellular locations: representative images from cellsCC-BY-NC-SA · From the PDF
Figure 4 Confocal images of A549 cells treated for 24 h with 100 μg/ml green 50nm silica nanoparticles in serum free medium. In red: EEA1 staining of the early endosomes or LAMP1 staining of the lysosomes (secondary antibody conjugated with Alexa-647). A and B) low magnification. C and D) enlarged details of single cells in the same conditions. Blue: DAPI stained nuclei.
Figure 4 Confocal images of A549 cells treated for 24 h with 100 μg/ml green 50nm silica nanoparticles in serum free medium. In red: EEA1 staining of the early endosomes or LAMP1 staining of the lysosomes (secondary antibody conjugated with Alexa-647). A and B) low magnification. C and D) enlarged details of single cells in the same conditions. Blue: DAPI stained nuclei.CC-BY-NC-SA · From the PDF
Figure 5 Adhesion of 50 nm silica nanoparticles on the cell membrane of A549 cells in serum free (SF) and complete medium (cMEM). Nanoparticles (100 µg/ml) were exposed to A549 cells at 4°C for different times in order to prevent nanoparticle uptake (as shown in Figures 5 and S3) and allow them to adhere on the cell membrane. The nanoparticle dispersion was then replaced by complete medium without nanoparticles and cells were warmed to 37°C and grown for further 3 h prior to flow cytometry assessment of internalised nanoparticle levels (see Methods for more details). Error bars are the standard deviation of the mean cell fluorescence intensity averaged over 3 replicas. B) The results in cMEM from panel A and the background fluorescence of untreated control cells.
Figure 5 Adhesion of 50 nm silica nanoparticles on the cell membrane of A549 cells in serum free (SF) and complete medium (cMEM). Nanoparticles (100 µg/ml) were exposed to A549 cells at 4°C for different times in order to prevent nanoparticle uptake (as shown in Figures 5 and S3) and allow them to adhere on the cell membrane. The nanoparticle dispersion was then replaced by complete medium without nanoparticles and cells were warmed to 37°C and grown for further 3 h prior to flow cytometry assessment of internalised nanoparticle levels (see Methods for more details). Error bars are the standard deviation of the mean cell fluorescence intensity averaged over 3 replicas. B) The results in cMEM from panel A and the background fluorescence of untreated control cells.CC-BY-NC-SA · From the PDF
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CC-BY-NC-SA · From the PDF

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Metadata

Authors
Lesniak, A., Fenaroli, F., Monopoli, M. P., Åberg, C., Dawson, K. A., & Salvati, A.
Venue
ACS Nano
Publisher
American Chemical Society (ACS)
Published
2012-06-29
Volume
6
Issue
7
Pages
5845-5857
ISSN
  • 1936-0851
  • 1936-086X
DOI
10.1021/nn300223w
Cited by
1,020
References
2
Language
en

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