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Signalling of DNA damage and cytokines across cell barriers exposed to nanoparticles depends on barrier thickness

Nature Nanotechnology volume 6, pages 824–833 (2011)Cite this article

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Abstract

The use of nanoparticles in medicine is ever increasing, and it is important to understand their targeted and non-targeted effects. We have previously shown that nanoparticles can cause DNA damage to cells cultured below a cellular barrier without crossing this barrier. Here, we show that this indirect DNA damage depends on the thickness of the cellular barrier, and it is mediated by signalling through gap junction proteins following the generation of mitochondrial free radicals. Indirect damage was seen across both trophoblast and corneal barriers. Signalling, including cytokine release, occurred only across bilayer and multilayer barriers, but not across monolayer barriers. Indirect toxicity was also observed in mice and using ex vivo explants of the human placenta. If the importance of barrier thickness in signalling is a general feature for all types of barriers, our results may offer a principle with which to limit the adverse effects of nanoparticle exposure and offer new therapeutic approaches.

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Figure 1: Signalling varies with thickness of BeWo barrier.
Figure 2: Oxidative stress, mitochondrial ROS or changes in oxygen level lead to DNA-damaging signalling from bilayered but not predominantly monolayered BeWo barriers.
Figure 3: Changes in BeWo barriers after nanoparticle exposure.
Figure 4: DNA damage in human embryonic stem cells below nanoparticle-exposed BeWo barriers.
Figure 5: DNA damage, in vivo and ex vivo, across bilayered but not monolayered barriers.
Figure 6: Signalling occurs through a bilayered but not monolayered corneal barrier.

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Acknowledgements

Advice and discussion was kindly provided by A. Halestrap (University of Bristol). P.W.A. and P.J.G. acknowledge support from the Wellcome Trust and from the Medical Research Council (project grant). The authors wish to thank O. Beaumont and L. Teoh for their help in calcium imaging experiments.

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Author notes

  1. A. Sood and S. Salih: These authors contributed equally to the practical work

Authors and Affiliations

  1. Bristol Musculoskeletal Research Unit, Clinical Science at North Bristol University of Bristol, Avon Orthopaedic Centre, Southmead Hospital, Bristol, BS10 5NB, UK

    A. Sood, S. Salih, L. Lacharme-Lora, M. Parry, B. Hardiman, R. Keehan & C. P. Case

  2. Department of Ophthalmology, UPMC Eye and Ear Institute, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, USA

    D. Roh

  3. Institut für Molekularbiologie (Tumorforschung) Institute of Molecular Biology (Tumor Research), Universitätsklinikum Essen University Hospital Essen Hufelandstrasse, Hufelandstrasse 55 55 45122, Essen, Germany

    R. Grummer & E. Winterhager

  4. Department of Biomedical Science, The University of Sheffield, Western Bank, Sheffield, S10 2TN, UK

    P. J. Gokhale & P. W. Andrews

  5. Department of Histopathology, Southmead Hospital, Bristol, BS10 5NB, UK

    C. Abbott

  6. Maternal and Fetal Health Research Group, The University of Manchester, 5th Floor Research, St Mary's Hospital, Manchester, M13 9WL, UK

    K. Forbes, M. Westwood & J. D. Aplin

  7. Institute of Molecular and Cellular Biology, University of Leeds, Mount Preston Street, Leeds, West Yorkshire, LS2 9JT, UK

    E. Ingham & I. Papageorgiou

  8. Ophthalmology Department, School of Clinical Sciences University of Bristol, Bristol Eye Hospital, Lower Maudlin Street, Bristol, BS1 2LX, UK

    M. Berry, J. Liu & A. D. Dick

  9. School of Cellular and Molecular Medicine University of Bristol Office, Medical Sciences Building, University Walk, Clifton, Bristol, BS8 1TD, UK

    R. J. Garland & N. Williams

  10. Department of Cancer Studies and Molecular Medicine, University of Leicester, University Road, Leicester, LE1 7RH, UK

    R. Singh

  11. The Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford, OX3 9DS, UK

    A. K. Simon

  12. Department of Medicine, Centre for Endocrine & Diabetes Sciences, Cardiff University School of Medicine, UHW Main Building, Heath Park, Cardiff, CF14 4XN, UK

    M. Lewis & J. Ham

  13. Department of Medical Genetics, Haematology & Pathology, Cardiff University School of Medicine, Henry Wellcome Building, Heath Park, Cardiff, CF14 4XN, UK

    L. Roger & D. M. Baird

  14. School of Clinical Sciences, University of Bristol, Dorothy Hodgkin Building, Whitson Street, Bristol, BS1 3NY, UK

    L. A. Crompton & M. A. Caldwell

  15. Dermatological Sciences, Institute of Cellular Medicine, The Medical School, Newcastle University, Newcastle, NE2 4HH, UK

    H. Swalwell & M. Birch-Machin

  16. Faculty of Life Sciences, University of Manchester, Manchester, M13 9PL, UK

    G. Lopez-Castejon

  17. School of Medical Sciences, University Walk, Bristol, BS8 1TD, UK

    A. Randall

  18. Cardiac Physiology, School of Clinical Sciences University of Bristol, Level 7, Bristol Royal Infirmary, Marlborough Street, Bristol, BS2 8HW, UK

    H. Lin & M-S. Suleiman

  19. Department of Infection, Immunity & Biochemistry, Cardiff University School of Medicine, Wales Heart Research Institute, Heath Park, Cardiff, CF14 4XN, UK

    W. H. Evans

  20. National Heart & Lung Institute, Imperial College, Emmanuel Kaye Building 1b, Manresa Road, London, SW3 6LR, UK

    R. Newson

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  1. A. Sood
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Contributions

C.P.C., A.S. and S.S. conceived and designed the experiments. A.S., S.S., L.L., B.H., R.G., E.W., D.R., P.G., C.A., K.F., M.W., I.P., M.B., R.J., M.S., D.B., G.L-C., L.C., J.L., K.S., M.L., L.R., H.S., M.B.M., A.R. and H.L. performed the experiments. C.P.C., R.N., D.R., P.G., N.M., J.A., A.R., M.B.M., J.H. and K.S. analysed the data. M.S., W.H.E., I.P. and E.I. contributed materials and analysis tools. C.P.C. wrote the paper.

Corresponding author

Correspondence to C. P. Case.

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Sood, A., Salih, S., Roh, D. et al. Signalling of DNA damage and cytokines across cell barriers exposed to nanoparticles depends on barrier thickness. Nature Nanotech 6, 824–833 (2011). https://doi.org/10.1038/nnano.2011.188

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