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In vitro chondrocyte toxicity following long-term, high-dose exposure to Gd-DTPA and a novel cartilage-targeted MR contrast agent

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Abstract

Objective

To determine the concentrations exhibiting toxicity of a cartilage-targeted magnetic resonance imaging contrast agent compared with gadopentetate dimeglumine (Gd-DT-PA) in chondrocyte cultures.

Materials and methods

A long-term Swarm rat chondrosarcoma chondrocyte-like cell line was exposed for 48 h to 1.0–20 mM concentrations of diaminobutyl-linked nitroxide (DAB4-DLN) citrate, 1.0–20 mM Gd-DTPA, 1.0 μM staurosporine (positive control), or left untreated. Cell appearance, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays of metabolic activity, quantitative PicoGreen assays of DNA content, and calcein-AM viability assays were compared.

Results

At 1.0–7.5 mM, minimal decrease in cell proliferation was found for both agents. At all doses of both agents, cell culture appearances were similar after 24 h of treatment. At the higher doses, differences in cell culture appearance were found after 48 h of treatment, with dose-dependent declines in chondrocyte populations for both agents. Concentration-dependent declines in DNA content and calcein fluorescence were found after 48 h of treatment, but beginning at a lower dose of DAB4-DLN citrate than Gd-DTPA. Dose-dependent decreases in MTT staining (cell metabolism) were apparent for both agents, but larger effects were evident at a lower dose for DAB-DLN citrate. Poor MTT staining of cells exposed for 48 h to 20 mM DAB4-DLN citrate probably indicates dead or dying cells.

Conclusion

The minimal effect of the long-term exposure of model chondrocyte cell cultures to DAB4-DLN citrate and Gd-DTPA concentrations up to 7.5 mM (3x typical arthrographic administration) is supporting evidence that these doses are acceptable for MR arthrography. The findings are reassuring given that the experimental exposure to the contrast agents at sustained concentrations was much longer than when used clinically.

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Author information

Authors and Affiliations

  1. Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, 9500 Euclid Avenue, Cleveland, OH, 44195, USA

    Sharon Midura, Carl S. Winalski & Ronald J. Midura

  2. Imaging Institute, A21, Cleveland Clinic, 9500 Euclid Avenue, Cleveland, OH, 44195, USA

    Erika Schneider & Carl S. Winalski

  3. Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, Baltimore, MD, 21201, USA

    Gerald M. Rosen

  4. NitroSci Pharmaceuticals, New Berlin, WI, 53151, USA

    Erika Schneider & Gerald M. Rosen

Authors
  1. Sharon Midura
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  2. Erika Schneider
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  3. Gerald M. Rosen
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  4. Carl S. Winalski
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  5. Ronald J. Midura
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Corresponding author

Correspondence to Carl S. Winalski.

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Conflicts of interest

E. Schneider and G. Rosen are minority shareholders of NitroSci Pharmaceuticals, LLC. E. Schneider is the Secretary/Treasurer and G. Rosen the Vice-President of NitroSci Pharmaceuticals, LLC, with no monetary compensation. NitroSci is the patent holder for the dendrimer-linked nitroxide contrast agents. C. Winalski has a family member who is a minority shareholder and officer of NitroSci Pharmaceuticals, LLC.

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This work had no external funding source.

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Midura, S., Schneider, E., Rosen, G.M. et al. In vitro chondrocyte toxicity following long-term, high-dose exposure to Gd-DTPA and a novel cartilage-targeted MR contrast agent. Skeletal Radiol 46, 23–33 (2017). https://doi.org/10.1007/s00256-016-2502-8

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  • DOI: https://doi.org/10.1007/s00256-016-2502-8

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