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Synthesis, Purification, Characterization, and Imaging of Cy3-Functionalized Fluorescent Silver Nanoparticles in 2D and 3D Tumor Models

  • Jessica Swanner
  • Ravi SinghEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1790)

Abstract

Silver nanoparticles (AgNPs) have a high affinity for sulfhydryl (thiol) groups, which can be exploited for functionalization with various tracking and targeting moieties. Here, we describe how to reliably and reproducibly functionalize AgNPs with the fluorescent moiety cyanine3-polyethelyne glycol (5000 molecular weight)-thiol (Cy3-PEG5000-SH). We also demonstrate how to purify and characterize Cy3-functionalized AgNPs (Cy3-AgNPs). Additionally, we describe how these Cy3-AgNPs can be imaged in 2D and 3D tumor models, providing insight into cellular localization and diffusion through a tumor spheroid, respectively.

Key words

Silver Nanomaterials Fluorescence Imaging 3D tumor models 

Notes

Acknowledgments

This work was supported in part by grant R00CA154006 (RS) from the National Institutes of Health, pilot funds from the Comprehensive Cancer Center of Wake Forest University supported by NCI CCSG P30CA012197, and by start-up funds from the Wake Forest School of Medicine Department of Cancer Biology. JS was supported in part by training grant T32CA079448 from the National Institutes of Health.

References

  1. 1.
    Chaloupka K, Malam Y, Seifalian AM (2010) Nanosilver as a new generation of nanoproduct in biomedical applications. Trends Biotechnol 28(11):580–588CrossRefPubMedGoogle Scholar
  2. 2.
    Gurunathan S et al (2013) Cytotoxicity of biologically synthesized silver nanoparticles in MDA-MB-231 human breast cancer cells. Biomed Res Int 2013:535796CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Jeyaraj M et al (2013) Biogenic silver nanoparticles for cancer treatment: an experimental report. Colloids Surf B Biointerfaces 106:86–92CrossRefPubMedGoogle Scholar
  4. 4.
    Liu JH et al (2012) TAT-modified nanosilver for combating multidrug-resistant cancer. Biomaterials 33(26):6155–6161CrossRefPubMedGoogle Scholar
  5. 5.
    Swanner J et al (2015) Differential cytotoxic and radiosensitizing effects of silver nanoparticles on triple-negative breast cancer and non-triple-negative breast cells. Int J Nanomedicine 10:3937–3953PubMedPubMedCentralGoogle Scholar
  6. 6.
    Liu PD et al (2013) Silver nanoparticles: a novel radiation sensitizer for glioma? Nanoscale 5(23):11829–11836CrossRefPubMedGoogle Scholar
  7. 7.
    Locatelli E et al (2014) Targeted delivery of silver nanoparticles and alisertib: in vitro and in vivo synergistic effect against glioblastoma. Nanomedicine 9(6):839–849CrossRefPubMedGoogle Scholar
  8. 8.
    Sharma S et al (2014) Silver nanoparticles impregnated alginate-chitosan-blended Nanocarrier induces apoptosis in human Glioblastoma cells. Adv Healthc Mater 3(1):106–114CrossRefPubMedGoogle Scholar
  9. 9.
    Miura N, Shinohara Y (2009) Cytotoxic effect and apoptosis induction by silver nanoparticles in HeLa cells. Biochem Biophys Res Commun 390(3):733–737CrossRefPubMedGoogle Scholar
  10. 10.
    Kawata K, Osawa M, Okabe S (2009) In vitro toxicity of silver nanoparticles at noncytotoxic doses to HepG2 human Hepatoma cells. Environ Sci Technol 43(15):6046–6051CrossRefPubMedGoogle Scholar
  11. 11.
    Beer C et al (2012) Toxicity of silver nanoparticles-nanoparticle or silver ion? Toxicol Lett 208(3):286–292CrossRefPubMedGoogle Scholar
  12. 12.
    Guo DW et al (2014) The cellular uptake and cytotoxic effect of silver nanoparticles on chronic myeloid Leukemia cells. J Biomed Nanotechnol 10(4):669–678CrossRefPubMedGoogle Scholar
  13. 13.
    Guo D et al (2013) Anti-leukemia activity of PVP-coated silver nanoparticles via generation of reactive oxygen species and release of silver ions. Biomaterials 34(32):7884–7894CrossRefPubMedGoogle Scholar
  14. 14.
    Shrivas K, Wu HF (2008) Applications of silver nanoparticles capped with different functional groups as the matrix and affinity probes in surface-assisted laser desorption/ionization time-of-flight and atmospheric pressure matrix-assisted laser desorption/ionization ion trap mass spectrometry for rapid analysis of sulfur drugs and biothiols in human urine. Rapid Commun Mass Spectrom 22(18):2863–2872CrossRefPubMedGoogle Scholar
  15. 15.
    Liau SY et al (1997) Interaction of silver nitrate with readily identifiable groups: relationship to the antibacterial action of silver ions. Lett Appl Microbiol 25(4):279–283CrossRefPubMedGoogle Scholar
  16. 16.
    Lynch I et al (2007) The nanoparticle - protein complex as a biological entity; a complex fluids and surface science challenge for the 21st century. Adv Colloid Interf Sci 134-35:167–174CrossRefGoogle Scholar
  17. 17.
    Roa W et al (2012) Pharmacokinetic and toxicological evaluation of multi-functional thiol-6-fluoro-6-deoxy-D-glucose gold nanoparticles in vivo. Nanotechnology 23(37):10CrossRefGoogle Scholar
  18. 18.
    Oberdorster G (2010) Safety assessment for nanotechnology and nanomedicine: concepts of nanotoxicology. J Intern Med 267(1):89–105CrossRefPubMedGoogle Scholar
  19. 19.
    Berti L et al (2010) Maximization of loading and stability of ssDNA:iron oxide nanoparticle complexes formed through electrostatic interaction. Langmuir 26(23):18293–18299CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Mattheolabakis G et al (2014) Pegylation improves the pharmacokinetics and bioavailability of small-molecule drugs hydrolyzable by esterases: a study of phospho-ibuprofen. J Pharmacol Exp Ther 351(1):61–66CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Cancer BiologyWake Forest School of MedicineWinston SalemUSA
  2. 2.Comprehensive Cancer Center of Wake Forest Baptist Medical CenterWinston SalemUSA

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