Design of Dense Brush Conformation Bearing Gold Nanoparticles as Theranostic Agent for Cancer

  • Nihan Verimli
  • Ayşegül Demiral
  • Hülya Yılmaz
  • Mustafa Çulha
  • S. Sibel ErdemEmail author


Dense brush conformation–bearing theranostic agents are emerging as drug delivery systems due to their higher ability to escape from reticuloendothelial system uptake which prolongs their in vivo circulation time. With the aim of developing dual therapy agent, 13-nm gold nanoparticles’ (AuNPs) surfaces were coated with different amounts of polyethylene glycol (PEG) (SH-PEG-NH2) to obtain dense brush conformation–bearing theranostic agents. Among the 14 different theranostic agent candidates prepared, the one hosting 1819 PEG per particle was selected as the most promising theranostic agent candidate based on structural conformation, stability, size, zeta potential, hemocompatibility, cell inhibition, and cell death pathway towards MCF-7 cell line. To test drug delivery efficiency of the developed PEGylated AuNP and to improve efficacy of the treatment, apoptotic peptide (AP) was covalently conjugated to NH2 terminus of the PEG in various ratios to yield AuNP-AP conjugate. Among the prepared conjugates, the one having 1 nmol of peptide per milliliter of AuNP yielded the most promising results based on the same criteria as employed for PEGylated AuNPs. Besides, incorporation of AP to AuNP returned in superior efficacy of AP since it was possible to achieve 50% cell death with 1000 times less amount of AP alone.


Apoptosis Apoptotic peptide Breast cancer Drug delivery Gold nanoparticles Hemocompatibility Polyethylene glycol Theranostic agent 



Apoptotic peptide


Bovine serum albumin


American Type Culture Collection


Gold nanoparticle


Computed tomography


Distance between two PEG attachments


Dynamic light scattering


Dulbecco’s modified Eagle’s medium


Dimethyl sulfoxide


5,5′-Dithiobis-(2-nitrobenzoic acid)


1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide


Enhanced permeability and retention


Fluorescence-activated cell sorting


Fetal bovine serum


Food and drug administration


Half maximal inhibitory concentration


Lactate dehyrdogenase


(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide






Phosphate-buffered saline


Polydispersity index


Polyethylene glycol




Reticuloendothelial system


Flory radius


Number of PEG per nanoparticles’ surface area


Surface-enhanced Raman scattering




Surface plasmon resonance


Transmission electron microscopy




Water-soluble tetrazolium salt


Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Ethical Issues

Istanbul Medipol University Non-Invasive Clinical Research Ethical Committee with issue number 10840098-604.01.01.-E.16355.

Supplementary material

12010_2019_3151_MOESM1_ESM.docx (92 kb)
ESM 1 (DOCX 91 kb)


  1. 1.
    Cho, K., Wang, X., Nie, S., Chen, Z., & Shin, D. M. (2008). Therapeutic nanoparticles for drug delivery in cancer. Clinical Cancer Research, 14(5), 1310–1316. Scholar
  2. 2.
    Conde, J., Dias, J. T., GrazÃo, V., Moros, M., Baptista, P. V., & de la Fuente, J. M. (2014). Revisiting 30 years of biofunctionalization and surface chemistry of inorganic nanoparticles for nanomedicine. Frontiers in Chemistry, 2(July), 1–27.
  3. 3.
    Dreaden, E. C., Austin, L. A., Mackey, M. A., & El-Sayed, M. A. (2012). Size matters: gold nanoparticles in targeted cancer drug delivery. Therapeutic Delivery, 3(4), 457–478. Scholar
  4. 4.
    Ma, A., & Czechowska, E. (2018). Immobilization of recombinant human catalase on gold and silver nanoparticles. Applied Biochemistry and Biotechnology, 185(3), 717–735. Scholar
  5. 5.
    Fernandez-fernandez, A., Manchanda, R., & Mcgoron, A. J. (2011). Theranostic applications of nanomaterials in cancer: drug delivery, image-guided therapy, and multifunctional platforms. Applied Biochemistry and Biotechnology, 165(7–8), 1628–1651. Scholar
  6. 6.
    Singh, P., Pandit, S., Mokkapati, V. R. S. S., Garg, A., Ravikumar, V., & Mijakovic, I. (2018). Gold nanoparticles in diagnostics and therapeutics for human cancer. International Journal of Molecular Sciences, 19(7). Scholar
  7. 7.
    Cormode, D. P., & Fayad, Z. A. (2011). Nanoparticle contrast agents for CT: their potential and the challenges that lie ahead, 3, 263–266.Google Scholar
  8. 8.
    Chatterjee, D. K., Diagaradjane, P., & Krishnan, S. (2011). Nanoparticle-mediated hyperthermia in cancer therapy. Therapeutic Delivery, 2(8), 1001–1014. Scholar
  9. 9.
    Pissuwan, D., Niidome, T., & Cortie, M. B. (2011). The forthcoming applications of gold nanoparticles in drug and gene delivery systems. Journal of Controlled Release, 149(1), 65–71. Scholar
  10. 10.
    Jokerst, J. V., Lobovkina, T., Zare, R. N., & Gambhir, S. S. (2011). Nanoparticle PEGylation for imaging and therapy. Nanomedicine, 6(4), 715–728. Scholar
  11. 11.
    Gupta, A. K., & Gupta, M. (2005). Synthesis and surface engineering of iron oxide nanoparticles for biomedical applications. Biomaterials, 26(18), 3995–4021. Scholar
  12. 12.
    Amani, A., Kabiri, T., Shafiee, S., & Hamidi, A. (2019). Preparation and characterization of PLA-PEG-PLA/PEI/DNA nanoparticles for improvement of transfection efficiency and controlled release of DNA in gene delivery systems. Iranian Journal of Pharmaceutical Research, 18(1), 125–141.PubMedGoogle Scholar
  13. 13.
    Leopold, L. F., Tódor, I. S., Diaconeasa, Z., Rugină, D., Ştefancu, A., Leopold, N., & Coman, C. (2017). Assessment of PEG and BSA-PEG gold nanoparticles cellular interaction. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 532(July), 70–76. Scholar
  14. 14.
    Su, L., Shu, T., Wang, Z., Cheng, J., Xue, F., Li, C., & Zhang, X. (2013). Biosensors and bioelectronics immobilization of bovine serum albumin-protected gold nanoclusters by using polyelectrolytes of opposite charges for the development of the reusable fluorescent Cu2+-sensor. Biosensors and Bioelectronic, 44, 16–20. Scholar
  15. 15.
    Conde, J., Ambrosone, A., Sanz, V., Hernandez, Y., Marchesano, V., Tian, F., Child, H., Berry, C. C., Ibarra, M. R., Baptista, P. V., Tortiglione, C., & de la Fuente, J. M. (2012). Design of multifunctional gold nanoparticles for in vitro and in vivo gene silencing. ACS Nano, 6(9), 8316–8324. Scholar
  16. 16.
    Paciotti, G. F., Kingston, D. G. I., & Tamarkin, L. (2006). Colloidal gold nanoparticles: a novel nanoparticle platform for developing multifunctional tumor-targeted drug delivery vectors (Vol. 54, pp. 47–54). Scholar
  17. 17.
    Sadzuka, Y., Kishi, K., Hirota, S., Sonobe, T. (2003). Effect of polyethyleneglycol (PEG) chain on cell uptake of PEG-modified liposomes, 13(2), 157–172. Scholar
  18. 18.
    Lipka, J., Semmler-behnke, M., Sperling, R. A., Wenk, A., Takenaka, S., Schleh, C., et al. (2010). Biomaterials biodistribution of PEG-modified gold nanoparticles following intratracheal instillation and intravenous injection. Biomaterials, 31(25), 6574–6581. Scholar
  19. 19.
    Suk, J. S., Xu, Q., Kim, N., Hanes, J., & Ensign, L. M. (2016). PEGylation as a strategy for improving nanoparticle-based drug and gene delivery. Advanced Drug Delivery Reviews, 99(Pt A), 28–51. Scholar
  20. 20.
    Peter, John, H. J. T. Cooper. (1951). A study of the nucleation and growth process in the synthesis of colloidal gold. Discussions of the Faraday Society, 55(c), 55–75. CrossRefGoogle Scholar
  21. 21.
    Murdock, R. C., Braydich-Stolle, L., Schrand, A. M., Schlager, J. J., & Hussain, S. M. (2008). Characterization of nanomaterial dispersion in solution prior to in vitro exposure using dynamic light scattering technique. Toxicological Sciences, 101(2), 239–253. Scholar
  22. 22.
    Zhang, X. D., Wu, D., Shen, X., Liu, P. X., Yang, N., Zhao, B., et al. (2011). Size-dependent in vivo toxicity of PEG-coated gold nanoparticles. International Journal of Nanomedicine, 6, 2071–2081. Scholar
  23. 23.
    Bartczak, D., & Kanaras, A. G. (2011). Preparation of peptide-functionalized gold nanoparticles using one pot EDC/Sulfo-NHS coupling. Langmuir, 27(16), 10119–10123. Scholar
  24. 24.
    Vijayakumar, S., & Ganesan, S. (2012). In vitro cytotoxicity assay on gold nanoparticles with different stabilizing agents. Journal of Nanomaterials, 2012, 1–9. Scholar
  25. 25.
    Steckiewicz, K. P., Barcinska, E., Malankowska, A., Zauszkiewicz–Pawlak, A., Nowaczyk, G., Zaleska-Medynska, A., & Inkielewicz-Stepniak, I. (2019). Impact of gold nanoparticles shape on their cytotoxicity against human osteoblast and osteosarcoma in in vitro model. Evaluation of the safety of use and anti-cancer potential. Journal of Materials Science: Materials in Medicine, 30(2), 1–15. Scholar
  26. 26.
    Chen, C., Cheng, Y. C., Yu, C. H., Chan, S. W., Cheung, M. K., & Yu, P. H. F. (2008). In vitro cytotoxicity, hemolysis assay, and biodegradation behavior of biodegradable poly(3-hydroxybutyrate)-poly(ethylene glycol)-poly(3-hydroxybutyrate) nanoparticles as potential drug carriers. Journal of Biomedical Materials Research - Part A, 87(2), 290–298. Scholar
  27. 27.
    Rahme, K., Chen, L., Hobbs, R. G., Morris, M. A., O’Driscoll, C., & Holmes, J. D. (2013). PEGylated gold nanoparticles: polymer quantification as a function of PEG lengths and nanoparticle dimensions. RSC Advances, 3(17), 6085–6094. Scholar
  28. 28.
    Moghimi, S. M., & Szebeni, J. (2003). Stealth liposomes and long circulating nanoparticles: critical issues in pharmacokinetics, opsonization and protein-binding properties, 42, 463–478. CrossRefGoogle Scholar
  29. 29.
    Urcan, E., Haertel, U., Styllou, M., Hickel, R., Scherthan, H., & Xaver, F. (2009). Real-time xCELLigence impedance analysis of the cytotoxicity of dental composite components on human gingival fibroblasts, 6, 51–58. CrossRefGoogle Scholar
  30. 30.
    Ju, P., Liang, R., Lee, Y., Zeng, Z., & Chuang, S. (2015). Differential cytotoxic effects of gold nanoparticles in different mammalian cell lines. Journal of Hazardous Materials, 264(2014), 303–312. Scholar
  31. 31.
    Dobrovolskaia, M. A., Clogston, J. D., Neun, B. W., Hall, J. B., Patri, A. K., & McNeil, S. E. (2008). Method for analysis of nanoparticle hemolytic properties in vitro. Nano Letters, 8(8), 2180–2187. Scholar
  32. 32.
    Iwasaki, T., Ishibashi, J., Tanaka, H., Sato, M., Asaoka, A., Taylor, D. M., & Yamakawa, M. (2009). Selective cancer cell cytotoxicity of enantiomeric 9-mer peptides derived from beetle defensins depends on negatively charged phosphatidylserine on the cell surface. Peptides, 30(4), 660–668. Scholar
  33. 33.
    Erdem, S. S., Obeidin, V. A., Yigitbasi, T., Tumer, S. S., & Yigit, P. (2018). Verteporfin mediated sequence dependent combination therapy against ovarian cancer cell line. Journal of Photochemistry and Photobiology B: Biology, 183(April), 266–274. Scholar
  34. 34.
    Öztaş, D. Y., Altunbek, M., Uzunoglu, D., Yllmaz, H., Cetin, D., Suludere, Z., & Culha, M. (2019). Tracing size and surface chemistry-dependent endosomal uptake of gold nanoparticles using surface-enhanced Raman scattering. Langmuir, 35(11), 4020–4028. Scholar
  35. 35.
    Farooq, M. U., Novosad, V., Rozhkova, E. A., Wali, H., Fateh, A. A., Neogi, P. B., et al. (2018). Gold nanoparticles-enabled efficient dual delivery of anticancer therapeutics to HeLa cells. Scientific Reports, 8(1), 1–12. Scholar
  36. 36.
    Albanese, A., & Chan, W. C. W. (2011). Effect of gold nanoparticle aggregation on cell uptake and toxicity, (7), 5478–5489.Google Scholar
  37. 37.
    Foroozandeh, P., & Aziz, A. A. (2018). Insight into cellular uptake and intracellular trafficking of nanoparticles. Nanoscale Research Letters, 13(1), 1–12.Google Scholar
  38. 38.
    Chow, E. K. H., & Ho, D. (2013). Cancer nanomedicine: from drug delivery to imaging. Science Translational Medicine, 5(216), 216), 1–216),12. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.International School of Medicine, Medical BiochemistryIstanbul Medipol UniversityIstanbulTurkey
  2. 2.Regenerative and Restorative Medical Research Center (REMER)IstanbulTurkey
  3. 3.School of Engineering and Natural Sciences, Biomedical EngineeringIstanbul Medipol UniversityIstanbulTurkey
  4. 4.Faculty of Engineering, Department of Genetics and BioengineeringYeditepe UniversityIstanbulTurkey

Personalised recommendations