Because of an increasing exposure to environmental and occupational nanoparticles (NPs), the potential risk of these materials for human health should be better assessed. Since one of the main routes of entry of NPs is via the lungs, it is of paramount importance to further characterize their impact on the respiratory system. Here, we have studied the uptake of fluorescently labeled SiO2 NPs (50 and 100 nm) by epithelial cells (NCI-H292) and alveolar macrophages (MHS) in the presence or absence of pulmonary surfactant. The quantification of NP uptake was performed by measuring cell-associated fluorescence using flow cytometry and spectrometric techniques in order to identify the most suitable methodology. Internalization was shown to be time and dose dependent, and differences in terms of uptake were noted between epithelial cells and macrophages. In the light of our observations, we conclude that flow cytometry is a more reliable technique for the study of NP internalization, and importantly, that the hydrophobic fraction of lung surfactant is critical for downregulating NP uptake in both cell types.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Blaaderen A, Vrij A (1992) Synthesis and characterization of colloidal dispersions of fluorescent. Monodisperse Silica Spheres, Langmuir, pp 2921–2931
Casals E, Pfaller T, Duschl A, Oostingh GJ, Puntes V (2010) Time evolution of the nanoparticle protein corona. ACS Nano 4:3623–3632
Chroneos ZC, Sever-Chroneos Z, Shepherd VL (2010) Pulmonary surfactant: an immunological perspective. Cell Physiol Biochem 25:13–26
Geiser M, Rothen-Rutishauser B, Kapp N, Schürch S, Kreyling W, Schulz H, Semmler M, Im Hof V, Heyder J, Gehr P (2005) Ultrafine particles cross cellular membranes by nonphagocytic mechanisms in lungs and in cultured cells. Environ Health Perspect 113:1555–1560
Geiser M, Casaulta M, Kupferschmid B, Schulz H, Semmler-Behnke M, Kreyling W (2008) The role of macrophages in the clearance of inhaled ultrafine titanium dioxide particles. Am J Respir Cell Mol Biol 38:371–376
Geiser M (2010) Update on macrophage clearance of inhaled micro- and nanoparticles. J Aerosol Med Pulm Drug Deliv 23:207–217
Kapralov AA, Feng WH, Amoscato AA, Yanamala N, Balasubramanian K, Winnica DE, Kisin ER, Kotchey GP, Gou P, Sparvero LJ, Ray P, Mallampalli RK, Klein-Seetharaman J, Fadeel B, Star A, Shvedova AA, Kagan VE (2012) Adsorption of surfactant lipids by single-walled carbon nanotubes in mouse lung upon pharyngeal aspiration. ACS Nano 6:4147–4156
Kruth HS (2002) Sequestration of agglomerated low-density lipoproteins by macrophages. Curr Opin Lipidol 13:483–488
Lesniak A, Fenaroli F, Monopoli MP, Aberg C, Dawson KA, Salvati A (2012) Effects of the presence or absence of a protein corona on silica nanoparticle uptake and impact on cells. ACS Nano 6:5845–5857
Lundqvist M, Stigler J, Cedervall T, Berggård T, Flanagan MB, Lynch I, Elia G, Dawson K (2011) The evolution of the protein corona around nanoparticles: a test study. ACS Nano 5:7503–7509
Lynch I, Cedervall T, Lundqvist M, Cabaleiro-Lago C, Linse S, Dawson KA (2007) The nanoparticle–protein complex as a biological entity; a complex fluids and surface science challenge for the 21st century. Adv Colloid Interface Sci 134–135:167–174
Lynch I, Salvati A, Dawson KA (2009) Protein–nanoparticle interactions: what does the cell see? Nat Nanotechnol 4:546–547
Mohamed BM, Verma NK, Prina-Mello A, Williams Y, Davies AM, Bakos G, Tormey L, Edwards C, Hanrahan J, Salvati A, Lynch I, Dawson K, Kelleher D, Volkov Y (2011) Activation of stress-related signalling pathway in human cells upon SiO2 nanoparticles exposure as an early indicator of cytotoxicity. J Nanobiotechnology 9:29
Mu Q, Hondow NS, Krzemi Ski L, Brown AP, Jeuken LJ, Routledge MN (2012) Mechanism of cellular uptake of genotoxic silica nanoparticles. Part Fibre Toxicol 9:29
Nel A, Xia T, Mädler L, Li N (2006) Toxic potential of materials at the nanolevel. Science 311:622–627
Oberdörster G, Oberdörster E, Oberdörster J (2005) Nanotoxicology: an emerging discipline evolving from studies of ultrafine particles. Environ Health Perspect 113:823–839
Rothen-Rutishauser BM, Schürch S, Haenni B, Kapp N, Gehr P (2006) Interaction of fine particles and nanoparticles with red blood cells visualized with advanced microscopic techniques. Environ Sci Technol 40:4353–4359
Roy I, Ohulchanskyy TY, Bharali DJ, Pudavar HE, Mistretta RA, Kaur N, Prasad PN (2005) Optical tracking of organically modified silica nanoparticles as DNA carriers: a nonviral, nanomedicine approach for gene delivery. Proc Natl Acad Sci U S A 102:279–284
Ruge CA, Schaefer UF, Herrmann J, Kirch J, Cañadas O, Echaide M, Pérez-Gil J, Casals C, Müller R, Lehr CM (2012) The interplay of lung surfactant proteins and lipids assimilates the macrophage clearance of nanoparticles. PLoS One 7:e40775
Schleh C, Mühlfeld C, Pulskamp K, Schmiedl A, Nassimi M, Lauenstein HD, Braun A, Krug N, Erpenbeck VJ, Hohlfeld JM (2009) The effect of titanium dioxide nanoparticles on pulmonary surfactant function and ultrastructure. Respir Res 10:90
Schleh C, Semmler-Behnke M, Lipka J, Wenk A, Hirn S, Schäffler M, Schmid G, Simon U, Kreyling WG (2012) Size and surface charge of gold nanoparticles determine absorption across intestinal barriers and accumulation in secondary target organs after oral administration. Nanotoxicology 6:36–46
Semmler-Behnke M, Takenaka S, Fertsch S, Wenk A, Seitz J, Mayer P, Oberdörster G, Kreyling WG (2007) Efficient elimination of inhaled nanoparticles from the alveolar region: evidence for interstitial uptake and subsequent reentrainment onto airways epithelium. Environ Health Perspect 115:728–733
Semmler-Behnke M, Kreyling WG, Lipka J, Fertsch S, Wenk A, Takenaka S, Schmid G, Brandau W (2008) Biodistribution of 1.4- and 18-nm gold particles in rats. Small 4:2108–2111
Sergent JA, Paget V, Chevillard S (2012) Toxicity and genotoxicity of Nano-SiO2 on human epithelial intestinal HT-29 cell line. Ann Occup Hyg 56(5):622–630
Wemhöner A, Frick M, Dietl P, Jennings P, Haller T (2006) A fluorescent microplate assay for exocytosis in alveolar type II cells. J Biomol Screen 11:286–295
Wu W, Samet JM, Peden DB, Bromberg PA (2010) Phosphorylation of p65 is required for zinc oxide nanoparticle-induced interleukin 8 expression in human bronchial epithelial cells. Environ Health Perspect 118:982–987
Xia T, Kovochich M, Liong M, Mädler L, Gilbert B, Shi H, Yeh JI, Zink JI, Nel AE (2008) Comparison of the mechanism of toxicity of zinc oxide and cerium oxide nanoparticles based on dissolution and oxidative stress properties. ACS Nano 2:2121–2134
This study was funded by the E.C. FP7 ENPRA (no. 228789) grant. Support for this study was also provided by Nanotrans (no. EST-2010/2/079) and TiSiTrans (no. PNR-EST-2010/2/79) grants. We acknowledge the confocal microscope platform in the Institute Jacques Monod, Paris, France. Authors would also like to thank to Dr Emmanuel Lopez (Cochin Hospital, Paris) for helping us to obtain the surfactant used in this study.
Responsible editor: Philippe Garrigues
About this article
Cite this article
Vranic, S., Garcia-Verdugo, I., Darnis, C. et al. Internalization of SiO2 nanoparticles by alveolar macrophages and lung epithelial cells and its modulation by the lung surfactant substitute Curosurf® . Environ Sci Pollut Res 20, 2761–2770 (2013). https://doi.org/10.1007/s11356-012-1436-5
- Mouse alveolar macrophages
- Flow cytometry
- Microplate reader
- Brilliant Black
- Silica nanoparticles