Radiolabeling of Preformed Niosomes with [99mTc]: In Vitro Stability, Biodistribution, and In Vivo Performance


Nanocarriers radiolabeled with [99mTc] can be used for diagnostic imaging and radionuclide therapy, as well as tracking their pharmacokinetic and biodistribution characteristics. Due to the advantages of niosomes as an ideal drug delivery system, in this study, the radiolabeling procedure of niosomes by [99mTc]-HMPAO complexes was investigated and optimized. Glutathione (GSH)-loaded niosomes were prepared using a thin-film hydration method. To label the niosomes with [99mTc], the preformed GSH-loaded niosomes were incubated with the [99mTc]-HMPAO complex and were characterized for particle size, size distribution, zeta potential, morphology, and radiolabeling efficiency (RE). The effects of GSH concentration, incubation time, incubation temperature, and niosomal composition on RE were investigated. The biodistribution profile and in vivo SPECT/CT imaging of the niosomes and free [99mTc]-HMPAO were also studied. Based on the results, all vesicles had nano-sized structure (160–235 nm) and negative surface charge. Among the different experimental conditions that were tested, including various incubation times, incubation temperatures, and GSH concentrations, the optimum condition that resulted in a RE of 92% was 200-mM GSH and 15-min incubation at 40°C. The in vitro release study in plasma showed that about 20% of radioactivity was released after 24 h, indicating an acceptable radiolabeling stability in plasma. The biodistribution of niosomes was clearly different from the free radiolabel. Niosomes carrying radionuclide were successfully used for tracking the in vivo disposition of these carriers and SPECT/CT imaging in rats. Furthermore, biodistribution studies in tumor-bearing mice revealed higher tumor accumulation of the niosomal formulation as compared with [99mTc]-HMPAO.

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Atomic force microscopy




Computed tomography


Diethylene triamine pentaacetic acid




Hexamethyl propylene amine oxime


Isomeric transition


Mononuclear phagocyte system


Phosphate buffered saline


Radiolabeling efficiency


Size-exclusion chromatography


Scanning electron microscopy


Single-photon emission computed tomography


Tween 60


Thin-layer chromatography


  1. 1.

    Goins B, Bao A, Phillips WT. Techniques for loading technetium-99m and rhenium-186/188 radionuclides into pre-formed liposomes for diagnostic imaging and radionuclide therapy. Methods Mol Biol. 2010;606:469–91.

    CAS  Article  Google Scholar 

  2. 2.

    Bartholomä MD, Louie AS, Valliant JF, Zubieta J. Technetium and gallium derived radiopharmaceuticals: comparing and contrasting the chemistry of two important radiometals for the molecular imaging era. Chem Rev. 2010;110(5):2903–20.

    Article  Google Scholar 

  3. 3.

    Ting G, Chang CH, Wang HE. Cancer nanotargeted radiopharmaceuticals for tumor imaging and therapy. Anticancer Res. 2009;29(10):4107–18.

    CAS  PubMed  Google Scholar 

  4. 4.

    Jain R, Dandekar P, Patravale V. Diagnostic nanocarriers for sentinel lymph node imaging. J Control Release. 2009;138(2):90–102.

    CAS  Article  Google Scholar 

  5. 5.

    Pereira MA, Mosqueira VC, Carmo VA, Ferrari CS, Reis EC, Ramaldes GA, et al. Biodistribution study and identification of inflammatory sites using nanocapsules labeled with (99m)Tc-HMPAO. Nucl Med Commun. 2009;30(9):749–55.

    Article  Google Scholar 

  6. 6.

    Goins BA. Radiolabeled lipid nanoparticles for diagnostic imaging. Expert Opin Med Diagn. 2008;2(7):853–73.

    CAS  Article  Google Scholar 

  7. 7.

    Phillips WT, Goins BA, Bao A. Radioactive liposomes. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2009;1(1):69–83.

    CAS  Article  Google Scholar 

  8. 8.

    Psimadas D, Bouziotis P, Georgoulias P, Valotassiou V, Tsotakos T, Loudos G. Radiolabeling approaches of nanoparticles with 99mTc. Contrast media & molecular imaging. 2013;8(4):333–9.

    CAS  Article  Google Scholar 

  9. 9.

    Dadashzadeh S, Mirahmadi N, Babaei MH, Vali AM. Peritoneal retention of liposomes: effects of lipid composition, PEG coating and liposome charge. J Control Release. 2010;148(2):177–86.

    CAS  Article  Google Scholar 

  10. 10.

    Bao A, Goins B, Klipper R, Negrete G, Phillips WT. Direct 99mTc labeling of pegylated liposomal doxorubicin (Doxil) for pharmacokinetic and non-invasive imaging studies. J Pharmacol Exp Ther. 2004;308(2):419–25.

    CAS  Article  Google Scholar 

  11. 11.

    Moghassemi S, Hadjizadeh A. Nano-niosomes as nanoscale drug delivery systems: an illustrated review. J Control Release. 2014;185:22–36.

    CAS  Article  Google Scholar 

  12. 12.

    Marianecci C, Di Marzio L, Rinaldi F, Celia C, Paolino D, Alhaique F, et al. Niosomes from 80s to present: the state of the art. Adv Colloid Interf Sci. 2014;205:187–206.

    CAS  Article  Google Scholar 

  13. 13.

    Arzani G, Haeri A, Daeihamed M, Bakhtiari-Kaboutaraki H, Dadashzadeh S. Niosomal carriers enhance oral bioavailability of carvedilol: effects of bile salt-enriched vesicles and carrier surface charge. Int J Nanomedicine. 2015;10:4797–813.

    CAS  PubMed  PubMed Central  Google Scholar 

  14. 14.

    Dilworth J, Parrott S. The biomedical chemistry of technetium and rhenium. Chem Soc Rev. 1998;27(1):43–55.

    CAS  Article  Google Scholar 

  15. 15.

    Phillips WT. Delivery of gamma-imaging agents by liposomes. Adv Drug Deliv Rev. 1999;37(1–3):13–32.

    CAS  Article  Google Scholar 

  16. 16.

    Korkmaz M, Özer A, Hincal A. DTPA niosomes in diagnostic imaging. Synthetic surfactant vesicles 2000. p. 248.

  17. 17.

    Laverman P, Boerman OC, Storm G. Radiolabeling of liposomes for scintigraphic imaging. Methods Enzymol: Elsevier; 2003. p. 234–48.

    Google Scholar 

  18. 18.

    Phillips WT, Rudolph AS, Goins B, Timmons JH, Klipper R, Blumhardt R. A simple method for producing a technetium-99m-labeled liposome which is stable in vivo. Int J Rad Appl Instrum B. 1992;19(5):539–47.

    CAS  Article  Google Scholar 

  19. 19.

    Rajera R, Nagpal K, Singh SK, Mishra DN. Niosomes: a controlled and novel drug delivery system. Biol Pharm Bull. 2011;34(7):945–53.

    CAS  Article  Google Scholar 

  20. 20.

    Sohrabi S, Haeri A, Mahboubi A, Mortazavi A, Dadashzadeh S. Chitosan gel-embedded moxifloxacin niosomes: an efficient antimicrobial hybrid system for burn infection. Int J Biol Macromol. 2016;85:625–33.

    CAS  Article  Google Scholar 

  21. 21.

    Mirahmadi N, Babaei MH, Vali AM, Daha FJ, Kobarfard F, Dadashzadeh S. 99m Tc-HMPAO-labeled liposomes: an investigation into the effects of some formulation factors on labeling efficiency and in vitro stability. Nucl Med Biol. 2008;35(3):387–92.

    CAS  Article  Google Scholar 

  22. 22.

    Khawar IA, Kim JH, Kuh HJ. Improving drug delivery to solid tumors: priming the tumor microenvironment. J Control Release. 2015;201:78–89.

    CAS  Article  Google Scholar 

  23. 23.

    Drummond DC, Noble CO, Hayes ME, Park JW, Kirpotin DB. Pharmacokinetics and in vivo drug release rates in liposomal nanocarrier development. J Pharm Sci. 2008;97(11):4696–740.

    CAS  Article  Google Scholar 

  24. 24.

    Song G, Wu H, Yoshino K, Zamboni WC. Factors affecting the pharmacokinetics and pharmacodynamics of liposomal drugs. J Liposome Res. 2012;22(3):177–92.

    CAS  Article  Google Scholar 

  25. 25.

    Blankenberg FG, Kinsman SL, Cohen BH, Goris ML, Spicer KM, Perlman SL, et al. Brain uptake of Tc99m-HMPAO correlates with clinical response to the novel redox modulating agent EPI-743 in patients with mitochondrial disease. Mol Genet Metab. 2012;107(4):690–9.

    CAS  Article  Google Scholar 

  26. 26.

    Bonte FJ, Hynan L, Harris TS, White CL 3rd. TC-99m HMPAO brain blood flow imaging in the dementias with histopathologic correlation in 73 patients. Int J Mol Imaging. 2011;2011(409101):1–3.

    Article  Google Scholar 

  27. 27.

    Varga Z, Gyurko I, Paloczi K, Buzas EI, Horvath I, Hegedus N, et al. Radiolabeling of extracellular vesicles with (99m)Tc for quantitative in vivo imaging studies. Cancer Biother Radiopharm. 2016;31(5):168–73.

    CAS  Article  Google Scholar 

  28. 28.

    Allan RA, Sladen GE, Bassingham S, Lazarus C, Clarke SE, Fogelman I. Comparison of simultaneous 99mTc-HMPAO and 111In oxine labelled white cell scans in the assessment of inflammatory bowel disease. Eur J Nucl Med. 1993;20(3):195–200.

    CAS  Article  Google Scholar 

  29. 29.

    Song G, Petschauer JS, Madden AJ, Zamboni WC. Nanoparticles and the mononuclear phagocyte system: pharmacokinetics and applications for inflammatory diseases. Curr Rheumatol Rev. 2014;10(1):22–34.

    CAS  Article  Google Scholar 

  30. 30.

    Ernsting MJ, Murakami M, Roy A, Li SD. Factors controlling the pharmacokinetics, biodistribution and intratumoral penetration of nanoparticles. J Control Release. 2013;172(3):782–94.

    CAS  Article  Google Scholar 

  31. 31.

    Salmaso S, Caliceti P. Stealth properties to improve therapeutic efficacy of drug nanocarriers. J Drug Deliv. 2013;2013(374252):1–19.

    Article  Google Scholar 

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This research was supported by a grant from Shahid Beheshti University of Medical Sciences (SBMU), Tehran, Iran.

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Corresponding authors

Correspondence to Soraya Shahhosseini or Simin Dadashzadeh.

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All animal studies were approved by the ethics committee for animal experiments at the Shahid Beheshti University of Medical science, Tehran, Iran.

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The authors declare that they have no conflict of interest.

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Almasi, A., Shahhosseini, S., Haeri, A. et al. Radiolabeling of Preformed Niosomes with [99mTc]: In Vitro Stability, Biodistribution, and In Vivo Performance. AAPS PharmSciTech 19, 3859–3870 (2018).

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  • radiolabeling
  • biodistribution
  • niosome
  • SPECT/CT imaging
  • [99mTc]-HMPAO