In vitro and in vivo toxicity and histopathological evaluation of Gd(III)anionic Linear globular dendrimer second-generation G2-C595 nanoprobe

Document Type : Research Paper


1 Cancer Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

2 Iran Ministry of Health and Medical Education, Deputy Ministry for Education, Tehran, Iran

3 Cancer Institute Research Center, Tehran University of Medical Sciences, Tehran, Iran

4 Department of Pathology, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran

5 Department of Medical Nanotechnology, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran

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


Objective(s): Toxico-histopathological studies are used to assess the toxic impacts of nanoparticles in organism exposure. The present study aimed to evaluate the prospective nano-cytotoxicity impacts of Gd(III)-anionic linear globular dendrimer second-generation G2-C595 (Gd[III] dendrimer G2-C595) contrast nanoprobe in terms of the exposure of many nude mice organs and organisms. In addition, we assessed the potential of the Gd(III)-dendrimer G2-C595 nanoprobe as a novel magnetic resonance imaging (MRI) nano-contrast agent for the human breast cancer cell line (MCF-7) and human embryonic kidney cell line (HEK-293).
Materials and Methods: Gadolinium (Gd[III]) was loaded with dendrimer G2 and conjugated with the C595 monoclonal antibody to generate the Gd(III)-dendrimer G2-C595 to determine the impact on MUC1 beneficial cancer tumors. The cytotoxic effects of the Gd(III)-dendrimer G2-C595 nanoprobe on the HEK-293 cells were also investigated in-vitro and in-vivo. In addition, the Gd(III)-dendrimer G2-C595 nanoprobe was used on nude mice bearing the MCF-7 tumors to explore its specific activity against the in-vivo model of cancer.
Results: The Gd(III)-dendrimer G2-C595 contrast nanoprobes affected the cytotoxicity of MCF-7, and no in-vivo toxicity was induced in the HEK-293 cells, kidneys, heart, lungs, brain, liver tissues, and other organs.
Conclusion: According to the results, the Gd(III)-dendrimer G2 and Gd(III)-dendrimer G2-C595 induced no toxicity in the HEK-293 cells and heart, liver, and brain tissues of mice. In addition, the Gd(III)-dendrimer G2-C595 showed specific anti-action against the in-vivo tumor model. Therefore, the Gd(III)-dendrimer G2-C595 nanoprobe is highly recommended as a novel and effective MR contrast agent and antitumor carrier agent. Furthermore, the Gd(III)-dendrimer G2-C595 nano-sized probes demonstrated excellent biocompatibility and safety with no impact on normal organ functioning.


1.Yang Y, Qin Z, Zeng W, Yang T, Cao Y, Mei C, Kuang Y. Toxicity assessment of nanoparticles in various systems and organs. Nanotechnol Rev. 2017; 6(3): 279-289.
2.Vandhana S, Nithya M, Deepak A. Review on nano toxic effects in living organisms (mice and zebra fish). Int J Innov Res Sci Technol. 2015; 1: 134-137.
3.Hubbs AF, Sargent LM, Porter DW, Sager TM, Chen BT, Frazer DG, Castranova V, Sriram K, Nurkiewicz TR, Reynolds SH, Battelli LA. Nanotechnology: toxicologic pathology. Toxicol Pathol. 2013; 41: 395-409.
4.Jain A, Dubey S, Kaushik A, Tyagi AK. Dendrimer: a complete drug carrier. IJPSR. 2010; 1(4): 38-52.
5.Priya P, Sivabalan.Mand Jeyapragash R. Dendrimer: A Novel Polymer. Ijrpc. 2013; 3(2): 495-501.
6.Wu L, Ficker M, Christensen JB, Trohopoulos PN, Moghimi SM. Dendrimers in Medicine: Therapeutic Concepts and Pharmaceutical Challenges. Bioconjug Chem. 2015; 26(7): 1198-1211.
7.Noriega-Luna B, Godínez LA, Rodríguez FJ, Rodríguez A, Zaldívar-Lelo de Larrea G, Sosa-Ferreyra CF, Mercado-Curiel RF, Manríquez J, Bustos E. Applications of Dendrimers in Drug Delivery Agents, Diagnosis, Therapy, and Detection. J Nanomater. 2014; 39: 1-19.
8.Ye M, Qian Y, Tang J, Hu H, Sui M, Shen Y. Targeted biodegradable dendritic MRI contrast agent for enhanced tumor imaging. J Control Release. 2013; 169: 239-45.
9.Ornelas C, Pennell R, Liebes LF, Weck M. Construction of a well-defined multifunctional dendrimer for theranostics. Org Lett. 2011; 13(5): 976-979.
10.Majoros IJ, Williams CR, Baker JR Jr. Current dendrimer application in cancer diagnosis and therapy. Curr Top Med Chem. 2008; 8: 1165-1179.
11.Sherje AP, Jadhav M, Dravyakar BR, Kadam D. Dendrimers: A versatile nanocarrier for drug delivery and targeting. Int J Pharm. 2018; 548(1): 707-720.
12.Chauhan AS. Dendrimers for Drug Delivery. Molecules. 2018; 23(4): 938.
13.Kesharwani P, Jain K, Jain NK. Dendrimer as nanocarrier for drug delivery. Prog Polym Sci. 2013; 39(2): 268–307.
14.Paleos CM, Tsiourvas D, Sideratou Z, Tziveleka LA. Drug delivery using multifunctional dendrimers and hyperbranched polymers. Expert Opin Drug Del. 2010; 7(12): 1387-1398.
15.Gupta U, Dwivedi SK, Bid HK, Konwar R, Jain NK. Ligand anchored dendrimers based nanoconstructs for effective targeting to cancer cells. Int J Pharm. 2010; 393: 185-96.
16.Singh J, Jain K, Mehra NK, Jain NK. Dendrimers in anticancer drug delivery: mechanism of interaction of drug and dendrimers. Artif Cells Nanomed Biotechnol. 2016; 44(7): 1626-34.
17.Zheng Qiao, Xiangyang Shi. Dendrimer-based molecular imaging contrast agents. Prog Polym Sci. 2015; 44: 1-27.
18.Zhang S, Zheng Y, Fu DY, Li W, Wu Y, Li B, Wu L. Biocompatible supramolecular dendrimers bearing a gadolinium-substituted polyanionic core for MRI contrast agents. J Mater Chem B. 2017; 5: 4035-4043.
19.Mirzaei M, Mehravi B, Ardestani MS, Ziaee SA, Pourghasem P. In vitro Evaluation of Gd(III)-Anionic Linear Globular Dendrimer-Monoclonal Antibody:Potential Magnetic Resonance Imaging Contrast Agents for Prostate Cancer Cell Imaging. Mol Imaging Biol. 2015; 17(6): 770-776.
20.Mullen DG, Fang M, Desai A, Baker JR, Orr BG, Banaszak Holl MM. A Quantitative assessment of nanoparticle-ligand distributions: implications for targeted drug and imaging delivery in dendrimer conjugates. ACS Nano. 2010; 4(2): 657–670.
21.Klemm PJ, Floyd WC, Smiles DE, Frà chet JMJ, Raymond KN. Improving T1 and T2 magnetic resonance imaging contrast agents through the conjugation of an esteramide dendrimer to high water coordination Gd(III) hydroxypyridinone complexes. Contrast Media Mol Imaging. 2012; 7: 95-99.
22.Longmire MR, Ogawa M, Choyke PL, Kobayashi H. Dendrimers as high relaxivity MR contrast agents. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2014; 6(2): 155-162.
23.Rolland O, Turrin CT, Caminadeab AM, Majoral JP. Dendrimers and nanomedicine: multivalency in action. New J Chem. 2009; 33, 1809-1824.
24.Jing X, Liang H, Hao C, Yang X, Cui X. Overexpression of MUC1 predicts poor prognosis in patients with breast cancer. Oncol Rep . 2019; 1;41(2): 801-810.
25.Stergiou N, Nagel J, Pektor S, Heimes AS, Jäkel J, Brenner W, Schmidt M, Miederer M, Kunz H, Roesch F, Schmitt E. Evaluation of a novel monoclonal antibody against tumor-associated MUC1 for diagnosis and prognosis of breast cancer. Int J Med Sci. 2019; 16(9): 1188-1198.
26.Lacunza E, Baudis M, Colussi AG, Segal-Eiras A, Croce MV, Abba MC. MUC1 oncogene amplification correlates with protein overexpression in invasive breast carcinoma cells. Cancer Genet Cytogenet. 2010; 201(2): 102-110.
27.Mirzaei M, Esmaeil Akbari M, Mohagheghi MA, Ziaee SAM, Mohseni M. A Novel Biocompatible Nanoprobe Based on Lipoproteins for Breast Cancer Cell Imaging. Nanomed J. 2019; 6(3): 73-79.
28.Mirzaei M, Mohseni M, Iranpour Anaraki N, Mohammadi E, Safari S, Mehravi B, Ghasempour A, Pourdakan O. New nanoprobe for breast cancer cell imaging based on low-density lipoprotein. Artif Cells Nanomed Biotechnol. 2020; 48(1): 46-52.
29.Shahbazi-Gahrouei D, Moradi Khaniabadi P, Moradi Khaniabadi B, Shahbazi-Gahrouei S. Medical imaging modalities using nanoprobes for cancer diagnosis: A literature review on recent findings. J Res Med Sci. 2019; 24:38.
30.Ko YJ, Kim WJ, Kim K, Kwon IC. Advances in the strategies for designing receptor-targeted molecular imaging probes for cancer research. J Control Release. 2019; 10; 305: 1-17.
31.Rajamanickam K. Multimodal Molecular Imaging Strategies using Functionalized Nano Probes. J Nanotechnol Res. 2019; 1(2): 119-135.
32.Clough TJ, Jiang L, Wong KL, Long NJ. Ligand design strategies to increase stability of gadolinium-based magnetic resonance imaging contrast agents. Nat Commun. 2019; 29; 10(1): 1420.
33.Jeong Y, Hwang HS, Na K. Theranostics and contrast agents for magnetic resonance imaging. Biomater Res. 2018; 22(1): 20.
34.Carvalho A, Gonçalves MC, Corvo ML, Martins MBF. Development of New Contrast Agents for Imaging Function and Metabolism by Magnetic Resonance Imaging. Magn Reson Insights. 2017; 10:1-4.
35.Mirzaei M, Mohagheghi M, Shahbazi-Gahrouei D, Khatami A. Gd3+-Anionic Linear Globular Dendrimer-G2-C595 A Dual Novel Nanoprobe for MR Imaging and Therapeutic Agent: An In- vitro Study. J Biomol Res Ther. 2012; 1:103.
36.Mirzaei M, Shahbazi-Gahrouei D, Mohagheghi M. Synthesis and development of Gd(III)-ALGDG2-C595 as MR imaging contrast agent. JBNB. 2013; 4: 22-29.
37.Lanone S, Boczkowski J. Biomedical applications and potential health risks of nanomaterials: molecular mechanisms. Curr Mol Med. 2006; 6(6): 651-63.
38.Soto K, Garza KM, Murr LE. Cytotoxic effects of aggregated nanomaterials. Acta Biomater. 2007, 3: 351-358.
39.Elsaesser A, Howard CV. Toxicology of nanoparticles. Adv Drug Deliv Rev. 2012; 64(2): 129-37.
40.Weyermann J, Lochmann D, Zimmer A. A practical note on the use of cytotoxicity assays. Int J Pharm. 2005; 288: 369-376.
41.Wörle-Knirsch JM, Pulskamp K, Krug HF. Oops they did it again! Carbon nanotubes hoax scientists in viability assays. Nano Lett. 2006; 6: 1261-1268.
42.Mirzaei M, Mohagheghi M, Shahbazi-Gahrouei D, Khatami A. Novel Nanosized Gd3+-ALGD-G2-C595: In vivo Dual Selective MUC1 Positive Tumor Molecular MR Imaging and Therapeutic Agent. J Nanomed Nanotechol. 2012; 3: 147.