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Labelling of Granulocytes by Phagocytic Engulfment with 64Cu-Labelled Chitosan-Coated Magnetic Nanoparticles

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An Erratum to this article was published on 22 December 2011

Abstract

Purpose

The aim of the present work was to perform the labelling of granulocytes by their engulfment with chitosan-coated magnetic 64Cu nanoparticles (MNPs) in order to obtain a radiopharmaceutical suitable for dual imaging (PET–MRI) of inflammatory/infective diseases.

Procedures

Specimens of 5–20 mg MNPs were washed with saline–isotonic solution and recuperated by magnetic decantation; 15–58 μg Cu2+ (CuCl2·H2O) in 50 μl of acidified (pH 5.5) saline solution was added to the MNPs re-suspended saline–isotonic solution; 10 mg MNPs was allowed to react with 16 μg 64Cu [64Ni(p,n) at 12–9 MeV] followed by anion exchange chromatography with a specific activity of 56 MBq/μg. Pellets of granulocytes were obtained from peripheral blood; MNPs engulfment by granulocytes was obtained and granulocyte-engulfed viability was assessed by the trypan blue exclusion (TBE) test performed at 5 min, 2 h and 4 h; assessment of the release of 64Cu from labelled granulocytes in plasma was performed by measuring the radioactivity of both the cellular pellet and the supernatant solution.

Results

Our data showed the binding capacity of chitosan-coated MNPs for cationic metal. The amount of Cu+2 chelated captured per milligram of MNPs was constant and independent of the reagent concentrations. In all cases, more than 90% of the engulfed granulocytes were positive to the TBE test. The MNPs were localised within the cells.

Conclusion

In our in vitro model, MNPs are taken up by granulocytes through phagocytosis, whereas previously described methods were based on the use of a chelating agent that permit Cu to cross the cell membrane. Moreover, the 64Cu-engulfed granulocytes showed a high stability of up to 80% of retained radioactivity after 24 h of incubation.

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Acknowledgement

The authors are deeply indebted to the Italian Company ACOM, Montecosaro, Macerata (Italy), for providing us 64Cu and for having made available equipment and expertise for the use of the radioisotope.

Conflict of Interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Department of Gynecology, Obstetrics and Urology, Laboratory of Biochemistry of Sexual Hormones, ‘Sapienza’ University of Rome, Rome, Italy

    Alessandra Pala & Piera D’Elia

  2. Department of Radiological, Oncological and Anatomo-Pathological Sciences, ‘Sapienza’ University of Rome, Rome, Italy

    Mauro Liberatore, Valentina Megna & Marco Mastantuono

  3. Department of Nuclear Medicine, Hammersmith Hospital, London, UK

    Fabio Nepi & Adil Al-Nahhas

  4. Department of Radiology, Medical Physics, Nuclear Medicine, Service of Nuclear Medicine, PET/CT Centre, ‘Santa Maria della Misericordia’ Hospital, Via Tre Martiri, 140, 45100, Rovigo, Italy

    Domenico Rubello

  5. Department of Chemistry, Faculty of Scienze M.F.N., ‘Sapienza’ University of Rome, Rome, Italy

    Mario Barteri

Authors
  1. Alessandra Pala
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  2. Mauro Liberatore
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  3. Piera D’Elia
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  4. Fabio Nepi
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  5. Valentina Megna
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  6. Marco Mastantuono
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  7. Adil Al-Nahhas
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  8. Domenico Rubello
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Corresponding author

Correspondence to Domenico Rubello.

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Pala, A., Liberatore, M., D’Elia, P. et al. Labelling of Granulocytes by Phagocytic Engulfment with 64Cu-Labelled Chitosan-Coated Magnetic Nanoparticles. Mol Imaging Biol 14, 593–598 (2012). https://doi.org/10.1007/s11307-011-0526-y

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  • DOI: https://doi.org/10.1007/s11307-011-0526-y

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