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Sputtering-Enabled Intracellular X-Ray Photoelectron Spectroscopy (SEI-XPS): A Versatile Method To Analyze The Biological Fate Of Metal Nanoparticles Antonio Turco, Mauro Moglianetti, Stefania Corvaglia, Simona Rella, Tiziano Catelani, Roberto Marotta, Cosimino Malitesta, and Pier Paolo Pompa ACS Nano, Just Accepted Manuscript • DOI: 10.1021/acsnano.8b01612 • Publication Date (Web): 13 Jul 2018 Downloaded from http://pubs.acs.org on July 14, 2018
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ACS Nano
Sputtering-Enabled Intracellular X-Ray Photoelectron Spectroscopy (SEI-XPS): A Versatile Method To Analyze The Biological Fate Of Metal Nanoparticles Antonio Turcoa*‡, Mauro Moglianettib*‡, Stefania Corvagliab, Simona Rellaa, Tiziano Catelani c, Roberto Marottac, Cosimino Malitestaa*, and Pier Paolo Pompab,d* a
Dipartimento di Scienze e Tecnologie Biologiche e Ambientali (Di.S.Te.B.A.),
Università del Salento, via Monteroni, – 73100 Lecce, Italy b
Istituto Italiano di Tecnologia, Nanobiointeractions & Nanodiagnostics, Center for
Bio-Molecular Nanotechnologies, Via Barsanti – 73010 Arnesano (Lecce), Italy c
Istituto Italiano di Tecnologia, Electron microscopy laboratory, Nanochemistry
department, Via Morego 30 – 16163 Genova, Italy d
Istituto Italiano di Tecnologia, Nanobiointeractions & Nanodiagnostics, Via
Morego 30 – 16163 Genova, Italy
*Corresponding authors ‡
These authors contributed equally
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Abstract The investigation of the toxicological profile and biomedical potential of nanoparticles (NPs) requires deep understanding of their intracellular fate. Various techniques are usually employed to characterize NPs upon cellular internalization, including high-resolution optical and electron microscopies. Here, we show a versatile method, named sputtering-enabled intracellular XPS (SEI-XPS), proving that it is able to provide valuable information about the behavior of metallic NPs in culture media as well as within cells, directly measuring their internalization, stability/degradation, and oxidation state, without any preparative steps. The technique can also provide nanoscale vertical resolution along with semi-quantitative information about the cellular internalization of the metallic species. The proposed approach
is
easy-to-use
and
can
become
a
standard
technique
in
nanotoxicology/nanomedicine and in the rational design of metallic NPs. Two model cases were investigated: silver nanoparticles (AgNPs) and platinum nanoparticles (PtNPs) with same size and coating. We observed that, after 48 hours incubation, intracellular AgNPs were almost completely dissolved, forming nanoclusters as well as Ag-O, Ag-S, and AgCl complexes. On the other hand, PtNPs were resistant to the harsh endo-lysosomal environment, and only some surface oxidation was detected after 48 hours.
Keywords: nanoparticles, XPS, cellular fate, oxidation state, degradation, nanotoxicology, nanomedicine
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ACS Nano
The promises of nanomedicine to deliver safe and effective drugs require detailed comprehension of the intracellular fate of nanoparticles (NPs).1,2 After their internalization within endo-lysosomes, NPs are subjected to the action of several small molecules and proteins, a harsh acidic environment, and aggressive proteolytic enzymes.3 However, up to now, there have been limited experimental possibilities to precisely assess the intracellular fate of NPs rather than in simulated biological fluids, leaving many scientific questions unsolved. The main technique to quantify NP internalization is Inductively Coupled Plasma Mass Spectrometry/Atomic Emission spectroscopy (ICP-MS/AES)4 that has long been used to determine quantitatively inorganic NP content after digestion of a cell suspension.5–7 However, this destructive technique does not provide any information about the NP cellular localization, degradation/biopersistence, and oxidative state. Laser Ablation Inductively Coupled Plasma Mass Spectrometry (LA-ICP-MS)6,7 offers the additional possibility to determine the subcellular localization of metallic NPs,8 but also in this latter case the degradation and oxidation state of the NP components cannot be assessed, losing crucial data on their potential toxicity. Imaging techniques, such as Focused Ion Beam Scanning Electron Microscopy (FIB/SEM)9, dark field microscopy,10 and Surface Enhanced Raman Scattering (SERS) imaging,11,12 provide information on the cellular localization of NPs, although with no assessment of the oxidation state of the material and strong limitations in case of small NPs (
