Synergistic cellular toxicity and uptake effects of iodixanol conjugated to anionic linear globular dendrimer G2

Document Type: Research Paper


1 Department of Radiopharmacy, School of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran

2 School of Pharmacy, Tehran University of Medical Sciences, International Branch, Tehran, Iran


Objective(s): Early diagnosis of cancer using noninvasive imaging techniques has been discussed in several recent studies. The present study aimed to assess the synergistic effects of iodixanol-conjugated polyethylene glycol (PEG)-citrate (anionic linear globular) dendrimer G2 on MCF-7 breast cancer cells and human embryonic kidney 293 (HEK293) cells.
Materials and Methods: PEG-citrate dendrimer G2 was synthesized and purified. The product was characterized using atomic force microscopy (AFM), electron energy loss spectroscopy (EELS), dynamic light scattering (DLS). At the next stage, the product was conjugated to iodixanol, purified and lyophilized. The cytotoxic effects of the iodixanol, plain PEG-citrate dendrimer G2, and iodixanol-PEG-citrate dendrimer G2 complex were evaluated using methylthiazole-tetrazolium (MTT) assay on the MCF-7 and HEK293 cells. Inductively coupled plasma mass spectrometry (ICP MS) is a mass spectrometry technique, which applies inductively coupled plasma to ionize samples.
Results: According to the obtained results, the uptake of PEG-citrate dendrimer G2 iodixanol increased significantly compared to iodixanol alone (P<0.05), indicating the importance of lack of significant in-vitro toxicity. Moreover, in the particle size and higher negative zeta potential confirmed the loading of iodixanol in dendrimer G2. Increase, the loading of iodixanol in dendrimer was confirmed by the chemical shifts in HNMR.
Conclusion: Therefore, it was concluded that the addition of anionic linear globular dendrimer G2 to iodixanol affected the cellular uptake of the drug with no significant toxicity. Recent findings also confirmed that this novel complex could be applied as an effective cancer imaging agent for molecular biology and molecular imaging applications.


1. Siegel R L, Miller K D , Jemal A. Cancer statistics. CA Cancer J Clin. 2016; (66): 7-30.
2. Smith R A. Cancer screening in the United States, 2015: a review of current American cancer society guidelines and current issues in cancer screening. CA Cancer J Clin. 2015; (65): 30-54.
3. Imperiale T F. Multitarget stool DNA testing for colorectal-cancer screening. N Engl J Med. 2014; (370): 1287-1297.
4. Choi K. Effect of endoscopy screening on stage at gastric cancer diagnosis: results of the National Cancer Screening Programme in Korea. Br J Cancer. 2015; (112): 608-612.
5. Heitzer E, Ulz P , Geigl J B. Circulating tumor DNA as a liquid biopsy for cancer. Clin Chem. 2015; (61): 112-123.
6. Hussain T , Nguyen Q T. Molecular imaging for cancer diagnosis and surgery. Adv Drug Deliv Rev. 2014; (66): 90-100.
7. Chen H, Zhen Z, Todd T, Chu P K , Xie J. Nanoparticles for improving cancer diagnosis. Mater Sci Eng R Rep. 2013; (74): 35-69.
8. Sepúlveda-Crespo D, Gómez R, De La Mata F J, Jiménez J L , Muñoz-Fernández M Á. Polyanionic carbosilane dendrimer-conjugated antiviral drugs as efficient microbicides: Recent trends and developments in HIV treatment/therapy. Nanomedicine: nanotechnology, biology, and medicine. 2015; (11): 1481-1498.
9. Worley B V, Slomberg D L , Schoenfisch M H. Nitric oxide-releasing quaternary ammonium-modified poly (amidoamine) dendrimers as dual action antibacterial agents. Bioconjug Chem. 2014; (25): 918-927.
10. McNelles S A, Knight S D, Janzen N, Valliant J F , Adronov A. Synthesis, radiolabeling, and in vivo imaging of PEGylated high-generation polyester dendrimers. Biomacromolecules. 2015; (16): 3033-3041.
11. Luong D. PEGylated PAMAM dendrimers: Enhancing efficacy , mitigating toxicity for effective anticancer drug and gene delivery. Acta Biomater. 2016; (43): 14-29.
12. Bosman A, Janssen H , Meijer E. About dendrimers: structure, physical properties, and applications. Chem Rev. 1999; (99): 1665-1688.
13. Lee C C, MacKay J A, Fréchet J M , Szoka, F C. Designing dendrimers for biological applications. Nat Biotechnol. 2005; (23): 1517-1526.
14. Ardestani M S. Nanosilver based anionic linear globular dendrimer with a special significant antiretroviral activity. J Mater Sci Mater Med. 2015; (26): 179.
15. Mohammadzadeh P, Cohan R A, Ghoreishi S M, Bitarafan-Rajabi A , Ardestani M S. AS1411 Aptamer-Anionic Linear Globular Dendrimer G2-Iohexol Selective Nano-Theranostics. Sci Rep. 2017; (7).
16. Ghoreishi S M, Bitarafan-Rajabi A, Azar A D, Ardestani M S , Assadi. A Novel 99mTc-Radiolabeled Anionic Linear Globular PEG-Based Dendrimer-Chlorambucil: Non-Invasive Method for In-Vivo Biodistribution. Drug Res. 2017; (67): 149-155.
17. Abdoli A. Conjugated anionic PEG-citrate G2 dendrimer with multi-epitopic HIV-1 vaccine candidate enhance the cellular immune responses in mice. Artif Cells Nanomed Biotechnol. 2017; 1-7.
18. Chalmers N , Jackson R. Comparison of iodixanol and iohexol in renal impairment. Br J Radiol. 1999; (72): 701-703.
19. Heinrich M C, Häberle L, Müller V, Bautz W , Uder M. Nephrotoxicity of iso-osmolar iodixanol compared with nonionic low-osmolar contrast media: meta-analysis of randomized controlled trials. Radiol. 2009; (250): 68-86.
20. Stacul F. Contrast induced nephropathy: updated ESUR contrast media safety committee guidelines. Eur Radiol. 2011; (21): 2527-2541.
21. Berg K. J. Nephrotoxicity related to contrast media. Scan J Urol Nephrol. 2000; (34), 317-322.
22. Dillman J R, Strouse P J, Ellis J H, Cohan R H , Jan S C. Incidence and severity of acute allergic-like reactions to iv nonionic iodinated contrast material in children. American journal of roentgenology. 2007;(188): 1643-1647.
23. Haririan I. Anionic linear-globular dendrimer-cis-platinum (II) conjugates promote cytotoxicity in vitro against different cancer cell lines. Int J Nanomedicine. 20105; (63).
24. Namazi H, Adeli M. Dendrimers of citric acid and poly (ethylene glycol) as the new drug-delivery agents. Biomaterials. 2005; (26): 1175-1183.
25. Alavidjeh, M S. Anionic linear-globular dendrimers: biocompatible hybrid materials with potential uses in nanomedicine. J Mater Sci Mater Med. 2010; (21): 1121-1133.
26. Barzegar Behrooz A. Smart bomb AS1411 aptamer‐functionalized/PAMAM dendrimer nanocarriers for targeted drug delivery in the treatment of gastric cancer. Clin Exp Pharmacol Physiol. 2017; (44): 41-51.
27. Arokiaraj M C, Am Heart J. 2016.
28. Hainfeld J, Slatkin D, Focella T , Smilowitz H. Gold nanoparticles: a new X-ray contrast agent. Br J Radiol. 2006; (79): 248-253.
29. Kim J, Chhour P , Hsu J , Litt, H I , Ferrari V A , Popovtzer, R Cormode D P. Use of Nanoparticle Contrast Agents for Cell Tracking with Computed Tomography. Bioconjug Chem. 2017; (21): 1581–1597.