, Volume 32, Issue 1, pp 21–32 | Cite as

Copper complex with sulfamethazine and 2,2′-bipyridine supported on mesoporous silica microspheres improves its antitumor action toward human osteosarcoma cells: cyto- and genotoxic effects

  • Juan Fernando Cadavid-Vargas
  • Pablo Maximiliano Arnal
  • Ruth Dary Mojica Sepúlveda
  • Andrea Rizzo
  • Delia Beatriz Soria
  • Ana Laura Di VirgilioEmail author


Ideal drugs to cure cancer leave normal cells unharmed while selectively turning tumor cells unviable. Several copper complexes have been able to selectively slow down tumor proliferation. We hypothesized that Cu(smz)2(bipy)·H2O (1)—a copper-complex that has two ligands capable of interacting with DNA—would outperform Cu(smz)2(OH2)·2H2O (2), and also that supporting 1 on mesoporous silica spheres would decrease even further tumor cell viability in vitro. After exposing osteosarcoma cells (MG-63) and normal phenotype cells of bone origin (MC3T3-E1) to either complex, we studied their toxic effect and mechanisms of action. We determined cell viability (MTT assay) and quantified formation of reactive oxygen species (oxidation of DHR-123 to rhodamine). Moreover, we assessed genotoxicity from (i) formation of micronucleus (MN assay) and (ii) damage of DNA (Comet assay). After the exposure of 1 supported on silica spheres, we tested cell viability. Our results confirm our hypotheses: inhibition of tumor cells follows: supported 1 > dissolved 1 > 2. Future work that enhances the load of the complex exclusively in mesopores may improve the ability of 1 to further inhibit tumor cell viability.


Antitumor effect Copper(II) complexes Cytotoxicity Genotoxicity Mesoporous silica microspheres 



The work was supported by UNLP (11X/690), CONICET (PIP 0105), and ANPCyT (PICT 2014-2223, PICT 2014-2583 and PICT 2016-0508) from Argentina.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10534_2018_154_MOESM1_ESM.tif (282 kb)
Supplementary material 1 (TIFF 281 kb). Supplementary material Fig. 1. Electronic spectra of 1
10534_2018_154_MOESM2_ESM.png (36 kb)
Supplementary material 2 (PNG 35 kb). Supplementary material Fig. 2. FT- IR spectra of 1 (green), silica spheres (blue) and silica supported complex (red)
10534_2018_154_MOESM3_ESM.tif (93 kb)
Supplementary material 3 (TIFF 93 kb). Supplementary material Fig. 3. Thermal gravimetric analysis of the silica spheres (blue), 1 (green) and complex supported on silica spheres (red)
10534_2018_154_MOESM4_ESM.tif (466 kb)
Supplementary material 4 (TIFF 465 kb). Supplementary material Fig. 4. A. SEM image of silica spheres. B. complex supported on spheres. The arrow indicates a complex crystal. C. EDX spectra of silica supported 1
10534_2018_154_MOESM5_ESM.docx (14 kb)
Supplementary material 5 (DOCX 13 kb)


  1. Ahola M, Kortesuo P, Kangasniemi I, Kiesvaara J, Yli-Urpo A (2000) Silica xerogel carrier material for controlled release of toremifene citrate. Int J Pharm 195(1–2):219–227PubMedGoogle Scholar
  2. Anderson RJ, Groundwater PW, Todd A, Worsley AJ (2012) Sulfonamide antibacterial agents. Wiley, Chichester, pp 103–126Google Scholar
  3. Arjmand F, Muddassir M (2011) A mechanistic approach for the DNA binding of chiral enantiomeric L- and d-tryptophan-derived metal complexes of 1,2-DACH: cleavage and antitumor activity. Chirality 23(3):250–259PubMedGoogle Scholar
  4. Arnal PM, Schüth F, Kleitz F (2006) A versatile method for the production of monodisperse spherical particles and hollow particles: templating from binary core-shell structures. Chem Commun 11:1203–1205Google Scholar
  5. Azqueta A, Collins AR (2013) The essential comet assay: a comprehensive guide to measuring DNA damage and repair. Arch Toxicol 87(6):949–968PubMedGoogle Scholar
  6. Becco L, García-Ramos JC, Azuara LR, Gambino D, Garat B (2014) Analysis of the DNA interaction of copper compounds belonging to the Casiopeínas® antitumoral series. Biol Trace Elem Res 161(2):210–215PubMedGoogle Scholar
  7. Bharti C, Nagaich U, Pal AK, Gulati N (2015) Mesoporous silica nanoparticles in target drug delivery system: a review. Int J Pharm Investig 5(3):124–133PubMedPubMedCentralGoogle Scholar
  8. Boulsourani Z, Katsamakas S, Geromichalos GD, Psycharis V, Raptopoulou CP, Hadjipavlou-Litina D et al (2017) Synthesis, structure elucidation and biological evaluation of triple bridged dinuclear copper(II) complexes as anticancer and antioxidant/anti-inflammatory agents. Mater Sci Eng, C 76:1026–1040Google Scholar
  9. Buchtík R, Trávníček Z, Vančo J, Herchel R, Dvořák Z (2011) Synthesis, characterization, DNA interaction and cleavage, and in vitro cytotoxicity of copper(ii) mixed-ligand complexes with 2-phenyl-3-hydroxy-4(1H)-quinolinone. Dalton Trans 40(37):9404PubMedGoogle Scholar
  10. Buchtík R, Trávníček Z, Vančo J (2012) In vitro cytotoxicity, DNA cleavage and SOD-mimic activity of copper(II) mixed-ligand quinolinonato complexes. J Inorg Biochem 116:163–171PubMedGoogle Scholar
  11. Cadavid-Vargas J, Leon I, Etcheverry S, Santi E, Torre M, Di Virgilio A (2017) Copper(II) complexes with saccharinate and glutamine as antitumor agents: cytoand genotoxicity in human osteosarcoma cells. Anticancer Agents Med Chem 17(3):424–433PubMedGoogle Scholar
  12. Carter MT, Rodriguez M, Bard AJ (1989) Voltammetric studies of the interaction of metal chelates with DNA. 2. Tris-chelated complexes of cobalt(III) and iron(II) with 1,10-phenanthroline and 2,2′-bipyridine. J Am Chem Soc 111(24):8901–8911Google Scholar
  13. Chang D, Gao Y, Wang L, Liu G, Chen Y, Wang T et al (2016) Polydopamine-based surface modification of mesoporous silica nanoparticles as pH-sensitive drug delivery vehicles for cancer therapy. J Colloid Interface Sci 463:279–287PubMedGoogle Scholar
  14. Chen X, Tang L-J, Sun Y-N, Qiu P-H, Liang G (2010) Syntheses, characterization and antitumor activities of transition metal complexes with isoflavone. J Inorg Biochem 104(4):379–384PubMedGoogle Scholar
  15. Cusumano M, Di Pietro ML, Giannetto A, Vainiglia PA (2005) The intercalation to DNA of bipyridyl complexes of platinum(II) with thioureas. J Inorg Biochem 99(2):560–565PubMedGoogle Scholar
  16. Czekanska EM, Stoddart MJ, Richards RG, Hayes JS (2012) In search of an osteoblast cell model for in vitro research. Eur Cells Mater 24:1–17. Google Scholar
  17. Dong A, Ren N, Yang W, Wang Y, Zhang Y, Wang D et al (2003) Preparation of hollow zeolite spheres and three-dimensionally ordered macroporous zeolite monoliths with functionalized interiors. Adv Funct Mater 13(12):943–948Google Scholar
  18. Duff B, Reddy Thangella V, Creaven BS, Walsh M, Egan DA (2012) Anti-cancer activity and mutagenic potential of novel copper(II) quinolinone Schiff base complexes in hepatocarcinoma cells. Eur J Pharmacol 689(1–3):45–55PubMedGoogle Scholar
  19. Fenech M (2000) The in vitro micronucleus technique. Mutat Res Mol Mech Mutagen 455(1–2):81–95Google Scholar
  20. Ferrari MB, Bisceglie F, Pelosi G, Tarasconi P, Albertini R, Dall’Aglio PP et al (2004) Synthesis, characterization and biological activity of copper complexes with pyridoxal thiosemicarbazone derivatives. X-ray crystal structure of three dimeric complexes. J Inorg Biochem 98(2):301–312Google Scholar
  21. Filho JCC, Sarria ALF, Becceneri AB, Fuzer AM, Batalhão JR, da Silva CMP et al (2014) Copper (II) and 2,2′-bipyridine complexation improves chemopreventive effects of naringenin against breast tumor cells. PLoS ONE 9(9):e107058PubMedPubMedCentralGoogle Scholar
  22. Finn NA, Kemp ML (2012) Pro-oxidant and antioxidant effects of N-acetylcysteine regulate doxorubicin-induced NF-kappa B activity in leukemic cells. Mol BioSyst 8(2):650–662PubMedGoogle Scholar
  23. Galindo-Murillo R, Hernandez-Lima J, González-Rendón M, Cortés-Guzmán F, Ruíz-Azuara L, Moreno-Esparza R (2011) $π-Stacking between Casiopeinas® and DNA bases. Phys Chem Chem Phys 13(32):14510PubMedGoogle Scholar
  24. Galindo-Murillo R, García-Ramos JC, Ruiz-Azuara L, Cheatham TE, Cortés-Guzmán F, Cortés-Guzmán F (2015) Intercalation processes of copper complexes in DNA. Nucleic Acids Res 43(11):5364–5376PubMedPubMedCentralGoogle Scholar
  25. Gaur R, Choubey DK, Usman M, Ward BD, Roy JK, Mishra L (2017) Synthesis, structures, nuclease activity, cytotoxicity, DFT and molecular docking studies of two nitrato bridged homodinuclear (Cu-Cu, Zn-Zn) complexes containing 2,2′-bipyridine and a chalcone derivative. J Photochem Photobiol B Biol 173:650–660Google Scholar
  26. Gutiérrez L, Alzuet G, Borrás J, Castiñeiras A, Rodríguez-Fortea A, Ruiz E (2001) Copper(II) complexes with 4-amino-N-[4,6-dimethyl-2-pyrimidinyl]benzenesulfonamide. Synthesis, crystal structure, magnetic properties, EPR, and theoretical studies of a novel mixed μ-carboxylato NCN-bridged dinuclear copper compound. Inorg Chem 40(13):3089–3096PubMedGoogle Scholar
  27. Hossain GMG, Amoroso AJ, Banu A, Malik KMA (2007) Syntheses and characterisation of mercury complexes of sulfadiazine, sulfamerazine and sulfamethazine. Polyhedron 26(5):967–974Google Scholar
  28. Karlsson H, Fryknäs M, Strese S, Gullbo J, Westman G, Bremberg U et al (2017) Mechanistic characterization of a copper containing thiosemicarbazone with potent antitumor activity. Oncotarget 8(18):30217–30234PubMedPubMedCentralGoogle Scholar
  29. León IE, Cadavid-Vargas JF, Di Virgilio AL, Etcheverry S (2016) Vanadium, ruthenium and copper compounds: a new class of non-platinum Metallodrugs with anticancer activity. Curr Med Chem 12(2):309–316Google Scholar
  30. Lobana TS, Indoria S, Jassal AK, Kaur H, Arora DS, Jasinski JP (2014) Synthesis, structures, spectroscopy and antimicrobial properties of complexes of copper(II) with salicylaldehyde N-substituted thiosemicarbazones and 2,2′-bipyridine or 1,10-phenanthroline. Eur J Med Chem 76:145–154PubMedGoogle Scholar
  31. Lu J, Liong M, Zink JI, Tamanoi F (2007) Mesoporous silica nanoparticles as a delivery system for hydrophobic. Anticancer Drugs 3(8):1341–1346Google Scholar
  32. Mai Z, Chen J, Hu Y, Liu F, Fu B, Zhang H et al (2017) Novel functional mesoporous silica nanoparticles loaded with Vitamin E acetate as smart platforms for pH responsive delivery with high bioactivity. J Colloid Interface Sci 508:184–195PubMedGoogle Scholar
  33. Mansour AM, Mohamed RR (2015) Sulfamethazine copper(ii) complexes as antimicrobial thermal stabilizers and co-stabilizers for rigid PVC: spectroscopic, thermal, and DFT studies. RSC Adv 5(7):5415–5423Google Scholar
  34. Martínez-Carmona M, Lozano D, Colilla M, Vallet-Regí M (2016) Selective topotecan delivery to cancer cells by targeted pH-sensitive mesoporous silica nanoparticles. RSC Adv 6(56):50923–50932Google Scholar
  35. Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol 65(1–2):55–63Google Scholar
  36. Nordberg J, Arnér ESJ (2001) Reactive oxygen species, antioxidants, and the mammalian thioredoxin system. Free Radic Biol Med 31(11):1287–1312PubMedGoogle Scholar
  37. Oliveri V, Lanza V, Milardi D, Viale M, Maric I, Sgarlata C et al (2017) Amino- and chloro-8-hydroxyquinolines and their copper complexes as proteasome inhibitors and antiproliferative agents. Metallomics 9(10):1439–1446PubMedGoogle Scholar
  38. Öztürk F, Bulut İ, Bulut A (2015) Structural, spectroscopic, magnetic and electrochemical studies of monomer N-substituted-sulfanilamide copper (II) complex with 2,2′-bipyridine. Spectrochim Acta, Part A 138:891–899Google Scholar
  39. Pautke C, Schieker M, Tischer T, Kolk A, Neth P, Mutschler W, Milz S (2004) Characterization of osteosarcoma cell lines MG-63, Saos-2 and U-2 OS in comparison to human osteoblasts. Anticancer Res 24:3743–3748PubMedGoogle Scholar
  40. Piotrowska-Kirschling A, Drzeżdżon J, Kloska A, Wyrzykowski D, Chmurzyński L, Jacewicz D (2018) Antioxidant and cytoprotective activity of oxydiacetate complexes of cobalt(II) and nickel(II) with 1,10-phenantroline and 2,2′-bipyridine. Biol Trace Elem Res 185(1):244–251PubMedGoogle Scholar
  41. Radin S, Ducheyne P, Kamplain T, Tan BH (2001) Silica sol-gel for the controlled release of antibiotics. I. Synthesis, characterization, andin vitro release. J Biomed Mater Res 57(2):313–320PubMedGoogle Scholar
  42. Royall JA, Ischiropoulos H (1993) Evaluation of 2′,7′-Dichlorofluorescin and dihydrorhodamine 123 as fluorescent probes for intracellular H2O2 in cultured endothelial cells. Arch Biochem Biophys 302(2):348–355PubMedGoogle Scholar
  43. Rytkönen J, Miettinen R, Kaasalainen M, Lehto V-P, Salonen J, Närvänen A (2012) Functionalization of mesoporous silicon nanoparticles for targeting and bioimaging purposes. J Nanomater 2012:1–9Google Scholar
  44. Santini C, Pellei M, Gandin V, Porchia M, Tisato F, Marzano C (2014) Advances in copper complexes as anticancer agents. Chem Rev 114(1):815–862PubMedGoogle Scholar
  45. Seng H-L, Wang W-S, Kong S-M, Alan Ong H-K, Win Y-F, Abd Raja, Rahman RNZ et al (2012) Biological and cytoselective anticancer properties of copper(II)-polypyridyl complexes modulated by auxiliary methylated glycine ligand. Biometals 25(5):1061–1081PubMedGoogle Scholar
  46. Serment-Guerrero J, Cano-Sanchez P, Reyes-Perez E, Velazquez-Garcia F, Bravo-Gomez ME, Ruiz-Azuara L (2011) Genotoxicity of the copper antineoplastic coordination complexes casiopeinas. Toxicol. 25(7):1376–1384Google Scholar
  47. Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175(1):184–191PubMedGoogle Scholar
  48. Stopper H, Müller SO (1997) Micronuclei as a biological endpoint for genotoxicity: a minireview. Toxicol Vitro 11(5):661–667Google Scholar
  49. Tommasino J-B, Renaud FNR, Luneau D, Pilet G (2011) Multi-biofunctional complexes combining antiseptic copper(II) with antibiotic sulfonamide ligands: structural, redox and antibacterial study. Polyhedron 30(10):1663–1670Google Scholar
  50. Wang J, Wang Y, Liu Q, Yang L, Zhu R, Yu C et al (2016) Rational design of multifunctional dendritic mesoporous silica nanoparticles to load curcumin and enhance efficacy for breast cancer therapy. ACS Appl Mater Interfaces 8(40):26511–26523PubMedGoogle Scholar
  51. Xu F, Ding L, Tao W, Yang X, Qian H, Yao R (2016) Mesoporous-silica-coated upconversion nanoparticles loaded with vitamin B12 for near-infrared-light mediated photodynamic therapy. Mater Lett 167:205–208Google Scholar
  52. Zhang H, Thomas R, Oupicky D, Peng F (2007) Synthesis and characterization of new copper thiosemicarbazone complexes with an ONNS quadridentate system: cell growth inhibition, S-phase cell cycle arrest and proapoptotic activities on cisplatin-resistant neuroblastoma cells. J Biol Inorg Chem 13(1):47–55PubMedGoogle Scholar
  53. Zhang Q, Neoh KG, Xu L, Lu S, Kang ET, Mahendran R et al (2014) Functionalized mesoporous silica nanoparticles with mucoadhesive and sustained drug release properties for potential bladder cancer therapy. Langmuir 30(21):6151–6161PubMedGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.CEQUINOR (CONICET-UNLP), Facultad de Ciencias ExactasUniversidad Nacional de La PlataLa PlataArgentina
  2. 2.CETMIC (Centro de Tecnología de Recursos Minerales y Cerámica)M.B. GonnetArgentina
  3. 3.Facultad de Ciencias ExactasUniversidad Nacional de La PlataLa PlataArgentina

Personalised recommendations