Skip to content
Licensed Unlicensed Requires Authentication Published by De Gruyter May 8, 2015

Impact of food components during in vitro digestion of silver nanoparticles on cellular uptake and cytotoxicity in intestinal cells

  • Dajana Lichtenstein EMAIL logo , Johanna Ebmeyer , Patrick Knappe , Sabine Juling , Linda Böhmert , Sören Selve , Birgit Niemann , Albert Braeuning , Andreas F. Thünemann and Alfonso Lampen
From the journal Biological Chemistry


Because of the rising application of nanoparticles in food and food-related products, we investigated the influence of the digestion process on the toxicity and cellular uptake of silver nanoparticles for intestinal cells. The main food components – carbohydrates, proteins and fatty acids – were implemented in an in vitro digestion process to simulate realistic conditions. Digested and undigested silver nanoparticle suspensions were used for uptake studies in the well-established Caco-2 model. Small-angle X-ray scattering was used to estimate particle core size, size distribution and stability in cell culture medium. Particles proved to be stable and showed radii from 3.6 to 16.0 nm. Undigested particles and particles digested in the presence of food components were comparably taken up by Caco-2 cells, whereas the uptake of particles digested without food components was decreased by 60%. Overall, these findings suggest that in vivo ingested poly (acrylic acid)-coated silver nanoparticles may reach the intestine in a nanoscaled form even if enclosed in a food matrix. While appropriate for studies on the uptake into intestinal cells, the Caco-2 model might be less suited for translocation studies. Moreover, we show that nanoparticle digestion protocols lacking food components may lead to misinterpretation of uptake studies and inconclusive results.

Corresponding author: Dajana Lichtenstein, Federal Institute for Risk Assessment, Max-Dohrn-Straße 8-10, D-10589 Berlin, Germany, e-mail:


We thank Dr. Richard Palavinkas for excellent technical support with AAS measurements. Verena Holle (Free University Berlin) is kindly acknowledged for performing sample preparation for TEM. This work was supported by the German Research Foundation/Deutsche Forschungsgemeinschaft (DFG) (grant number: LA 1177/9-1).

Conflict of interest disclaimer: The authors declare no competing financial interest.


Artursson, P. and Karlsson, J. (1991). Correlation between oral drug absorption in humans and apparent drug permeability coefficients in human intestinal epithelial (Caco-2) cells. Biochem. Biophys. Res. Commun. 175, 880–885.10.1016/0006-291X(91)91647-USearch in Google Scholar

Beer, C., Foldbjerg, R., Hayashi, Y., Sutherland, D.S., and Autrup, H. (2012). Toxicity of silver nanoparticles – Nanoparticle or silver ion? Toxicol. Lett. 208, 286–292.Search in Google Scholar

Böhmert, L., Girod, M., Hansen, U., Maul, R., Knappe, P., Niemann, B., Weidner, S.M., Thünemann, A.F., and Lampen, A. (2014). Analytically monitored digestion of silver nanoparticles and their toxicity on human intestinal cells. Nanotoxicology 8, 631–642.10.3109/17435390.2013.815284Search in Google Scholar PubMed

Bouwmeester, H., Poortman, J., Peters, R.J., Wijma, E., Kramer, E., Makama, S., Puspitaninganindita, K., Marvin, H.J., Peijnenburg, A.A., and Hendriksen, P.J. (2011). Characterization of translocation of silver nanoparticles and effects on whole-genome gene expression using an in vitro intestinal epithelium coculture model. ACS Nano. 5, 4091–4103.10.1021/nn2007145Search in Google Scholar PubMed

Bowden, L.P., Royer, M.C., Hallman, J.R., Lewin-Smith, M., and Lupton, G.P. (2011). Rapid onset of argyria induced by a silver-containing dietary supplement. J. Cutan. Pathol. 38, 832–835.10.1111/j.1600-0560.2011.01755.xSearch in Google Scholar PubMed

Chaudhry, Q., Scotter, M., Blackburn, J., Ross, B., Boxall, A., Castle, L., Aitken, R., and Watkins, R. (2008). Applications and implications of nanotechnologies for the food sector. Food Addit. Contam. 25, 241–258.10.1080/02652030701744538Search in Google Scholar PubMed

des Rieux, A., Fievez, V., Théate, I., Mast, J., Préat, V., and Schneider, Y.-J. (2007). An improved in vitro model of human intestinal follicle-associated epithelium to study nanoparticle transport by M cells. Eur. J. Pharmaceut. Sci. 30, 380–391.10.1016/j.ejps.2006.12.006Search in Google Scholar PubMed

Desai, M.P., Labhasetwar, V., Amidon, G.L., and Levy, R.J. (1996). Gastrointestinal uptake of biodegradable microparticles: effect of particle size. Pharmaceut. Res. 13, 1838–1845.10.1023/A:1016085108889Search in Google Scholar

EFSA (2011). Guidance on the Risk assessment of the application of nanoscience and nanotechnologies in the food and feed chain. EFSA J. 9.Search in Google Scholar

Gerloff, K., Pereira, D.I., Faria, N., Boots, A.W., Kolling, J., Forster, I., Albrecht, C., Powell, J.J., and Schins, R.P. (2013). Influence of simulated gastrointestinal conditions on particle-induced cytotoxicity and interleukin-8 regulation in differentiated and undifferentiated Caco-2 cells. Nanotoxicology 7, 353–366.10.3109/17435390.2012.662249Search in Google Scholar PubMed PubMed Central

Glover, R.D., Miller, J.M., and Hutchison, J.E. (2011). Generation of metal nanoparticles from silver and copper objects: nanoparticle dynamics on surfaces and potential sources of nanoparticles in the environment. ACS Nano. 5, 8950–8957.10.1021/nn2031319Search in Google Scholar PubMed

Greulich, C., Kittler, S., Epple, M., Muhr, G., and Köller, M. (2009). Studies on the biocompatibility and the interaction of silver nanoparticles with human mesenchymal stem cells (hMSCs). Langenbeck’s Arch. Surg. 394, 495–502.10.1007/s00423-009-0472-1Search in Google Scholar PubMed

Hu, Y., Ge, J., Lim, D., Zhang, T., and Yin, Y. (2008). Size-controlled synthesis of highly water-soluble silver nanocrystals. J. Solid State Chem. 181, 1524–1529.10.1016/j.jssc.2008.02.028Search in Google Scholar

Kim, S., Choi, J.E., Choi, J., K.-H. Chung, Park, K., Yi, J., and Ryu, D.-Y. (2009). Oxidative stress-dependent toxicity of silver nanoparticles in human hepatoma cells. Toxicol. In Vitro 23, 1076–1084.10.1016/j.tiv.2009.06.001Search in Google Scholar PubMed

Kim, Y.S., Song, M.Y., Park, J.D., Song, K.S., Ryu, H.R., Chung, Y.H., Chang, H.K., Lee, J.H., Oh, K.H., Kelman, B.J., et al. (2010). Subchronic oral toxicity of silver nanoparticles. Part. Fibre Toxicol. 7, 20.10.1186/1743-8977-7-20Search in Google Scholar PubMed PubMed Central

Lesniak, A., Fenaroli, F., Monopoli, M.P., Åberg, C., Dawson, K.A., and Salvati, A. (2012). Effects of the presence or absence of a protein corona on silica nanoparticle uptake and impact on cells. ACS Nano. 6, 5845–5857.10.1021/nn300223wSearch in Google Scholar PubMed

Liu, J. and Hurt, R.H. (2010). Ion release kinetics and particle persistence in aqueous nano-silver colloids. Environ. Sci. Technol. 44, 2169–2175.10.1021/es9035557Search in Google Scholar PubMed

Liu, J., Sonshine, D.A., Shervani, S., and Hurt, R.H. (2010). Controlled release of biologically active silver from nanosilver surfaces. ACS Nano. 4, 6903–6913.10.1021/nn102272nSearch in Google Scholar PubMed PubMed Central

McCracken, C., Zane, A., Knight, D.A., Dutta, P.K., and Waldman, W.J. (2013). Minimal intestinal epithelial cell toxicity in response to short-and long-term food-relevant inorganic nanoparticle exposure. Chem. Res. Toxicol. 26, 1514–1525.10.1021/tx400231uSearch in Google Scholar PubMed

Monteiro-Riviere, N.A., Samberg, M.E., Oldenburg, S.J., and Riviere, J.E. (2013). Protein binding modulates the cellular uptake of silver nanoparticles into human cells: implications for in vitro to in vivo extrapolations? Toxicol. Lett. 220, 286–293.10.1016/j.toxlet.2013.04.022Search in Google Scholar PubMed PubMed Central

Munger, M.A., Radwanski, P., Hadlock, G.C., Stoddard, G., Shaaban, A., Falconer, J., Grainger, D.W., and Deering-Rice, C.E. (2014). In vivo human time-exposure study of orally dosed commercial silver nanoparticles. Nanomedicine 10, 1–9.10.1016/j.nano.2013.06.010Search in Google Scholar PubMed PubMed Central

Mwilu, S.K., El Badawy, A.M., Bradham, K., Nelson, C., Thomas, D., Scheckel, K.G., Tolaymat, T., Ma, L., and Rogers, K.R. (2013). Changes in silver nanoparticles exposed to human synthetic stomach fluid: effects of particle size and surface chemistry. Sci.Total Environ. 447, 90–98.10.1016/j.scitotenv.2012.12.036Search in Google Scholar PubMed

Pauw, B.R., Pedersen, J.S., Tardif, S., Takata, M., and Iversen, B.B. (2013). Improvements and considerations for size distribution retrieval from small-angle scattering data by Monte Carlo methods. J. Appl. Crystallogr. 46, 365–371.10.1107/S0021889813001295Search in Google Scholar PubMed PubMed Central

Peters, R., Kramer, E., Oomen, A.G., Herrera Rivera, Z.E., Oegema, G., Tromp, P.C., Fokkink, R., Rietveld, A., Marvin, H.J.P., Weigel, S., et al. (2012). Presence of nano-sized silica during in vitro digestion of foods containing silica as a food additive. ACS Nano. 6, 2441–2451.10.1021/nn204728kSearch in Google Scholar PubMed

Powell, J.J., Faria, N., Thomas-McKay, E., and Pele, L.C. (2010). Origin and fate of dietary nanoparticles and microparticles in the gastrointestinal tract. J. Autoimmun. 34, J226–J233.10.1016/j.jaut.2009.11.006Search in Google Scholar PubMed

Tiede, K., Boxall, A.B., Tear, S.P., Lewis, J., David, H., and Hassellöv, M. (2008). Detection and characterization of engineered nanoparticles in food and the environment. Food Addit. Contam. 25, 795–821.10.1080/02652030802007553Search in Google Scholar PubMed

von Goetz, N., Fabricius, L., Glaus, R., Weitbrecht, V., Gunther, D., and Hungerbuhler, K. (2013). Migration of silver from commercial plastic food containers and implications for consumer exposure assessment. Food Addit. Contam. Part A Chem. Anal. Control Expo. Risk Assess. 30, 612–20.10.1080/19440049.2012.762693Search in Google Scholar PubMed

Walczak, A.P., Fokkink, R., Peters, R., Tromp, P., Herrera Rivera, Z.E, Rietjens, I.M., Hendriksen, P.J., and Bouwmeester, H. (2013). Behaviour of silver nanoparticles and silver ions in an in vitro human gastrointestinal digestion model. Nanotoxicology 7, 1198–1210.10.3109/17435390.2012.726382Search in Google Scholar PubMed

Walczak, A.P., Kramer, E., Hendriksen, P.J., Tromp, P., Helsper, J.P., van der Zande, M., Rietjens, I.M., and Bouwmeester, H. (2014). Translocation of differently sized and charged polystyrene nanoparticles in in vitro intestinal cell models of increasing complexity. Nanotoxicology 5, 1–9.Search in Google Scholar

Supplemental Material:

The online version of this article (DOI: 10.1515/hsz-2015-0145) offers supplementary material, available to authorized users.

Received: 2015-1-16
Accepted: 2015-5-6
Published Online: 2015-5-8
Published in Print: 2015-11-1

©2015 by De Gruyter

Downloaded on 30.9.2023 from
Scroll to top button