Advertisement

Analytical and Bioanalytical Chemistry

, Volume 411, Issue 16, pp 3497–3506 | Cite as

High-resolution laser ablation inductively coupled plasma mass spectrometry used to study transport of metallic nanoparticles through collagen-rich microstructures in fibroblast multicellular spheroids

  • Akihiro ArakawaEmail author
  • Norbert Jakubowski
  • Sabine Flemig
  • Gunda Koellensperger
  • Mate Rusz
  • Daigo Iwahata
  • Heike Traub
  • Takafumi Hirata
Research Paper

Abstract

We have efficiently produced collagen-rich microstructures in fibroblast multicellular spheroids (MCSs) as a three-dimensional in vitro tissue analog to investigate silver (Ag) nanoparticle (NP) penetration. The MCS production was examined by changing the seeding cell number (500 to 40,000 cells) and the growth period (1 to 10 days). MCSs were incubated with Ag NP suspensions with a concentration of 5 μg mL−1 for 24 h. For this study, laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) was used to visualize Ag NP localization quantitatively. Thin sections of MCSs were analyzed by LA-ICP-MS with a laser spot size of 8 μm to image distributions of 109Ag, 31P, 63Cu, 66Zn, and 79Br. A calibration using a NP suspension was applied to convert the measured Ag intensity into the number of NPs present. The determined numbers of NPs ranged from 30 to 7200 particles in an outer rim of MCS. The particle distribution was clearly correlated with the presence of 31P and 66Zn and was localized in the outer rim of proliferating cells with a width that was equal to about twice the diameter of single cells. Moreover, abundant collagens were found in the outer rim of MCSs. For only the highest seeding cell number, NPs were completely captured at the outer rim, in a natural barrier reducing particle transport, whereas Eosin (79Br) used as a probe of small molecules penetrated into the core of MCSs already after 1 min of exposure.

Graphical abstract

Fibroblast MCS could build up the barrier only for nanoparticles

Keywords

Laser ablation inductively coupled plasma mass spectrometry Silver nanoparticles Fibroblast cells Multicellular spheroids 

Notes

Acknowledgements

We thank Konrad Löhr (Bundesanstalt für Materialforschung und -prüfung) for the support and training for the non-contact piezo-driven array spotter and Akvile Häckel (Charité Universitätsmedizin Berlin) for providing access to and support with using the cryomicrotome.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

216_2019_1827_MOESM1_ESM.pdf (1.2 mb)
ESM 1 (PDF 1191 kb)

References

  1. 1.
    Kessler R. Engineered nanoparticles in consumer products: understanding a new ingredient. Environ Health Perspect. 2011;119(3):A120–A5.CrossRefGoogle Scholar
  2. 2.
    Li W-R, Xie X-B, Shi Q-S, Zeng H-Y, OU-Yang Y-S, Chen Y-B. Antibacterial activity and mechanism of silver nanoparticles on Escherichia coli. Appl Microbiol Biotechnol. 2010;85(4):1115–22.CrossRefGoogle Scholar
  3. 3.
    Larguinho M, Baptista PV. Gold and silver nanoparticles for clinical diagnostics - from genomics to proteomics. J Proteome. 2012;75(10):2811–23.CrossRefGoogle Scholar
  4. 4.
    Wilkinson LJ, White RJ, Chipman JK. Silver and nanoparticles of silver in wound dressings: a review of efficacy and safety. J Wound Care. 2011;20(11):543–9.CrossRefGoogle Scholar
  5. 5.
    Reidy B, Haase A, Luch A, Dawson K, Lynch I. Mechanisms of silver nanoparticle release, transformation and toxicity: a critical review of current knowledge and recommendations for future studies and applications. Materials. 2013;6(6):2295.CrossRefGoogle Scholar
  6. 6.
    Aaron J, Travis K, Harrison N, Sokolov K. Dynamic imaging of molecular assemblies in live cells based on nanoparticle plasmon resonance coupling. Nano Lett. 2009;9(10):3612–8.CrossRefGoogle Scholar
  7. 7.
    Ando J, Fujita K, Smith NI, Kawata S. Dynamic SERS imaging of cellular transport pathways with endocytosed gold nanoparticles. Nano Lett. 2011;11(12):5344–8.CrossRefGoogle Scholar
  8. 8.
    Drescher D, Kneipp J. Nanomaterials in complex biological systems: insights from Raman spectroscopy. Chem Soc Rev. 2012;41(17):5780–99.CrossRefGoogle Scholar
  9. 9.
    Kneipp J, Kneipp H, Rice WL, Kneipp K. Optical probes for biological applications based on surface-enhanced Raman scattering from indocyanine green on gold nanoparticles. Anal Chem. 2005;77(8):2381–5.CrossRefGoogle Scholar
  10. 10.
    Huang X, Jain PK, El-Sayed IH, El-Sayed MA. Plasmonic photothermal therapy (PPTT) using gold nanoparticles. Lasers Med Sci. 2007;23(3):217.CrossRefGoogle Scholar
  11. 11.
    Nam J, Won N, Jin H, Chung H, Kim S. pH-induced aggregation of gold nanoparticles for photothermal cancer therapy. J Am Chem Soc. 2009;131(38):13639–45.CrossRefGoogle Scholar
  12. 12.
    Schneider G, Guttmann P, Heim S, Rehbein S, Mueller F, Nagashima K, et al. Three-dimensional cellular ultrastructure resolved by X-ray microscopy. Nat Methods. 2010;7:985.CrossRefGoogle Scholar
  13. 13.
    Guehrs E, Schneider M, Günther CM, Hessing P, Heitz K, Wittke D, et al. Quantification of silver nanoparticle uptake and distribution within individual human macrophages by FIB/SEM slice and view. J Nanobiotechnol. 2017;15(1):21.CrossRefGoogle Scholar
  14. 14.
    Alkilany AM, Murphy CJ. Toxicity and cellular uptake of gold nanoparticles: what we have learned so far? J Nanopart Res. 2010;12(7):2313–33.CrossRefGoogle Scholar
  15. 15.
    Krystek P, Ulrich A, Garcia CC, Manohar S, Ritsema R. Application of plasma spectrometry for the analysis of engineered nanoparticles in suspensions and products. J Anal At Spectrom. 2011;26(9):1701–21.CrossRefGoogle Scholar
  16. 16.
    Laux P, Tentschert J, Riebeling C, Braeuning A, Creutzenberg O, Epp A, et al. Nanomaterials: certain aspects of application, risk assessment and risk communication. Arch Toxicol. 2018;92(1):121–41.CrossRefGoogle Scholar
  17. 17.
    Sabine Becker J, Matusch A, Palm C, Salber D, Morton KA, Susanne Becker J. Bioimaging of metals in brain tissue by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) and metallomics. Metallomics. 2010;2(2):104–11.CrossRefGoogle Scholar
  18. 18.
    Van Acker T, Van Malderen SJM, Van Heerden M, McDuffie JE, Cuyckens F, Vanhaecke F. High-resolution laser ablation-inductively coupled plasma-mass spectrometry imaging of cisplatin-induced nephrotoxic side effects. Anal Chim Acta. 2016;945:23–30.CrossRefGoogle Scholar
  19. 19.
    Giesen C, Waentig L, Mairinger T, Drescher D, Kneipp J, Roos PH, et al. Iodine as an elemental marker for imaging of single cells and tissue sections by laser ablation inductively coupled plasma mass spectrometry. J Anal At Spectrom. 2011;26(11):2160–5.CrossRefGoogle Scholar
  20. 20.
    Drescher D, Giesen C, Traub H, Panne U, Kneipp J, Jakubowski N. Quantitative imaging of gold and silver nanoparticles in single eukaryotic cells by laser ablation ICP-MS. Anal Chem. 2012;84(22):9684–8.CrossRefGoogle Scholar
  21. 21.
    Scharlach C, Müller L, Wagner S, Kobayashi Y, Kratz H, Ebert M, et al. LA-ICP-MS allows quantitative microscopy of europium-doped iron oxide nanoparticles and is a possible alternative to ambiguous Prussian blue iron staining. J Biomed Nanotechnol. 2016;12(5):1001–10.CrossRefGoogle Scholar
  22. 22.
    Büchner T, Drescher D, Traub H, Schrade P, Bachmann S, Jakubowski N, et al. Relating surface-enhanced Raman scattering signals of cells to gold nanoparticle aggregation as determined by LA-ICP-MS micromapping. Anal Bioanal Chem. 2014;406(27):7003–14.CrossRefGoogle Scholar
  23. 23.
    Drescher D, Zeise I, Traub H, Guttmann P, Seifert S, Büchner T, et al. In situ characterization of SiO2 nanoparticle biointeractions using BrightSilica. Adv Funct Mater. 2014;24(24):3765–75.CrossRefGoogle Scholar
  24. 24.
    Theiner S, Schreiber-Brynzak E, Jakupec MA, Galanski M, Koellensperger G, Keppler BK. LA-ICP-MS imaging in multicellular tumor spheroids - a novel tool in the preclinical development of metal-based anticancer drugs. Metallomics. 2016;8(4):398–402.CrossRefGoogle Scholar
  25. 25.
    Theiner S, Van Malderen SJM, Van Acker T, Legin A, Keppler BK, Vanhaecke F, et al. Fast high-resolution laser ablation-inductively coupled plasma mass spectrometry imaging of the distribution of platinum-based anticancer compounds in multicellular tumor spheroids. Anal Chem. 2017;89(23):12641–5.CrossRefGoogle Scholar
  26. 26.
    Furukawa KS, Ushida T, Sakai Y, Kunii K, Suzuki M, Tanaka J, et al. Tissue-engineered skin using aggregates of normal human skin fibroblasts and biodegradable material. J Artif Organs. 2001;4(4):353–6.CrossRefGoogle Scholar
  27. 27.
    Priwitaningrum DL, Blondé J-BG, Sridhar A, van Baarlen J, Hennink WE, Storm G, et al. Tumor stroma-containing 3D spheroid arrays: a tool to study nanoparticle penetration. J Control Release. 2016;244:257–68.CrossRefGoogle Scholar
  28. 28.
    Jorgenson AJ, Choi KM, Sicard D, Smith KMJ, Hiemer SE, Varelas X, et al. TAZ activation drives fibroblast spheroid growth, expression of profibrotic paracrine signals, and context-dependent ECM gene expression. Am J Phys Cell Phys. 2017;312(3):C277–C85.CrossRefGoogle Scholar
  29. 29.
    Sapudom J, Pompe T. Biomimetic tumor microenvironments based on collagen matrices. Biomater Sci. 2018;6(8):2009–24.CrossRefGoogle Scholar
  30. 30.
    Emon B, Bauer J, Jain Y, Jung B, Saif T. Biophysics of tumor microenvironment and cancer metastasis - a mini review. Comput Struct Biotechnol J. 2018;16:279–87.CrossRefGoogle Scholar
  31. 31.
    Suzuki T, Sakata S, Makino Y, Obayashi H, Ohara S, Hattori K, et al. iQuant2: software for rapid and quantitative imaging using laser ablation-ICP mass spectrometry. Mass Spectrom. 2018;7(1):A0065-A.CrossRefGoogle Scholar
  32. 32.
    Schneider CA, Rasband WS, Eliceiri KW. NIH image to ImageJ: 25 years of image analysis. Nat Methods. 2012;9:671.CrossRefGoogle Scholar
  33. 33.
    Curcio E, Salerno S, Barbieri G, De Bartolo L, Drioli E, Bader A. Mass transfer and metabolic reactions in hepatocyte spheroids cultured in rotating wall gas-permeable membrane system. Biomaterials. 2007;28(36):5487–97.CrossRefGoogle Scholar
  34. 34.
    Drescher D. Spektro-Mikroskopische Charakterisierung von Nano-Bio-Wechselwirkungen in Zellen. PhD thesis. Humboldt University Berlin; 2016.Google Scholar
  35. 35.
    Drescher D, Guttmann P, Buchner T, Werner S, Laube G, Hornemann A, et al. Specific biomolecule corona is associated with ring-shaped organization of silver nanoparticles in cells. Nanoscale. 2013;5(19):9193–8.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Bundesanstalt für Materialforschung und -prüfung (BAM)BerlinGermany
  2. 2.Research Institute for Bioscience Products and Fine ChemicalsAjinomoto Co., Inc.Kawasaki-shiJapan
  3. 3.Spetec GmbHErdingGermany
  4. 4.Institute of Analytical ChemistryUniversity of ViennaViennaAustria
  5. 5.Cell Culture Facility, Institute of Inorganic ChemistryUniversity of ViennaViennaAustria
  6. 6.Geochemical Research CenterThe University of TokyoTokyoJapan

Personalised recommendations