Quantification of metal ion induced DNA damage with single cell array based assay

    Authors

    Y. Qiao;L. Y. Ma

    Comments

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    Abbreviated Journal Title

    Analyst

    Keywords

    TOTAL HIP-ARTHROPLASTY; COMET ASSAY; INDIVIDUAL CELLS; IN-VIVO; DEBRIS; REPAIR; TISSUE; FLUIDS; Chemistry, Analytical

    Abstract

    Under physiological and wear conditions, implanted orthopedic devices undergo undesired release of metal ions which cause DNA damage and inflammation of local tissue. However, individuals have personalized responses to identical devices due to varying susceptibility to DNA damage. The current one-size-fits-all approach is therefore not suitable to predict the response of patients to implanted devices. This paper describes a single cell array based method to quantify metal ion induced DNA damage that can potentially be used to predict the response to implanted devices in patients. Ions of several typical metals in implanted devices were used to treat human normal fibroblast cells. After patterning cells on a silicon substrate with cell-catching patches, cells were embedded in hydrogel and treated with alkaline buffer. Damaged DNAs diffuse out of the cell, and are stained to show a characteristic halo. All studied metal ions (Cu2+, Co2+, Ni2+, Cr2+, Fe2+, Al3+) induce DNA damage and have genotoxicity. Copper ions cause DNA damage at concentrations as low as 1 mu M. Cobalt and nickel ions damage DNA at 5 and 10 mu M, respectively. Aluminum, iron and chromium ions cause DNA damage at 50 mu M. The cytotoxicity assay shows that most ions, except cobalt and copper, are less toxic below 500 mu M. The fact that metal ions can cause genotoxicity at lower concentrations than that of cytotoxicity suggests: (1) a single cell based DNA damage assay is more sensitive than a membrane integrity based live/dead assay; and (2) metal ions preferentially induce DNA damage rather than cell membrane damage.

    Journal Title

    Analyst

    Volume

    138

    Issue/Number

    19

    Publication Date

    1-1-2013

    Document Type

    Article

    Language

    English

    First Page

    5713

    Last Page

    5718

    WOS Identifier

    WOS:000323722600026

    ISSN

    0003-2654

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