Measuring and modelling cell-to-cell variation in uptake of gold nanoparticles

Jeynes, Jonathan C. G.; Jeynes, C.; Merchant, Michael J; Kirkby, K.J.

doi:10.1039/c3an01406a

Cited by 40 publications

(46 citation statements)

References 32 publications

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“…The samples are placed into the vacuum chamber of a 2 MV tandem accelerator [13, 15]. A 2.5 MeV proton beam was focused to a 2 μ m spot size of about 300 pA. At this energy, the protons can penetrate the entire thickness of human cells (about 6 μ m) with very little energy loss.…”

Section: Methodsmentioning

confidence: 99%

“…The samples were scanned in 200 × 200 μ m regions for 5 h, with each region containing 10–30 cells. The energy spectrum of photons or scattered particles from each pixel is recorded (with OMDAQ) so that the data can be reconstructed and analysed offline using OMDAQ to manipulate the pixel data, and the DataFurnace code for quantification following the methods given in detail in [13]. …”

Section: Methodsmentioning

confidence: 99%

“…The details of the analysis procedure have been described in detail previously for single cells [13]. Briefly, using the signals from the proton induced x-ray emission (PIXE) detector the cells are imaged (particularly using the phosphorus signals) (see figures 2 and 3), and then can be quantitatively measured using the combined PIXE and elastic backscattered (EBS) signal using DataFurnace [16].…”

Section: Methodsmentioning

confidence: 99%

“…Nuclear microprobes have a long history of elemental mapping of biological tissues and cells on a micron scale with high sensitivity and accurate quantification available when x-ray and particle spectra are used self-consistently [11, 12]. Previously, we have used this technique to measure the absolute number of gold NPs in cancer cells [13] as well as the concentration of cis-platin within the nucleus of individual cells [14]. …”

Section: Introductionmentioning

confidence: 99%

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Direct quantification of rare earth doped titania nanoparticles in individual human cells

Jeynes

Palitsin

et al. 2016

Nanotechnology

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There are many possible biomedical applications for titania nanoparticles (NPs) doped with rare earth elements (REEs), from dose enhancement and diagnostic imaging in radiotherapy, to biosensing. However, there are concerns that the NPs could disintegrate in the body thus releasing toxic REE ions to undesired locations. As a first step, we investigate how accurately the Ti/REE ratio from the NPs can be measured inside human cells. A quantitative analysis of whole, unsectioned, individual human cells was performed using proton microprobe elemental microscopy. This method is unique in being able to quantitatively analyse all the elements in an unsectioned individual cell with micron resolution, while also scanning large fields of view. We compared the Ti/REE signal inside cells to NPs that were outside the cells, non-specifically absorbed onto the polypropylene substrate. We show that the REE signal in individual cells co-localises with the titanium signal, indicating that the NPs have remained intact. Within the uncertainty of the measurement, there is no difference between the Ti/REE ratio inside and outside the cells. Interestingly, we also show that there is considerable variation in the uptake of the NPs from cell-to-cell, by a factor of more than 10. We conclude that the NPs enter the cells and remain intact. The large heterogeneity in NP concentrations from cell-to-cell should be considered if they are to be used therapeutically.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Direct quantification of rare earth doped titania nanoparticles in individual human cells

Jeynes

Palitsin

et al. 2016

Nanotechnology

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“…This does not only refer to the obvious differences among the various cell types existing (e.g., plant vs. mammalian cells, prokaryotic vs. eukaryotic cells), but also on the possible differences in the uptake of NPs within one cell population because of different cell cycle phases. High fluctuations of intracellular load of NPs have been reported populations in in vitro cell culture studies using human alveolar epithelial [8] and human urinary bladder cell lines [9]. This is the reason why the prerequisite for a representative cell sample size is also of utmost importance in order to produce accurate and precise measurement values.…”

Section: General Considerations About Quantification Techniquesmentioning

confidence: 99%

Quantifying Nanoparticle Cellular Uptake: Which Method is Best?

Drašler

Vanhecke

Rodríguez‐Lorenzo

et al. 2017

Nanomedicine (Lond.)

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Keywords:As the range of engineered nanoparticles (NPs) designed as specific carriers increases, for example for cell targeting and drug delivery, the question on how many NPs are interacting or are taken up by cells is becoming increasingly important for any potential biomedical application. On one hand, the delivered dose of such NPs to the targeted cells is a key parameter in the assessment of their efficiency to perform the desired action (e.g., deliver the therapeutic substance or induce a specific effect), on the other hand, the assessment of intracellular NPs is crucial also from the safety aspect as NPs might come unintentionally in contact by untargeted cells. Particularly from the regulative perspective, it is important that reproducible and reliable analytical methods for the intracellular quantification of NPs are available at an early stage in the development in order to correlate the cell burden of NPs with their possible effects at a cellular level. Which method is the best?The authors approached this matter by questioning senior scientists in the field of nanoscience about their personal views on the prime method for quantifying intracellular NPs. Although -at first glance -this is a seemingly straightforward enquiry, their responses were rather ambiguous yet consistent: in a nutshell, they all concluded the method of choice for the quantification of NP uptake mainly depends on the research question, the available analytical devices as well as on the type of NPs of interest. As of that, it is not possible to recommend one specific technique that could be used for quantification of all the different NPs types which exist nowadays. As well known from the convincing evidence from the literature, physicochemical properties of NPs such as their size, shape, core material and surface functionalization have a strong impact on NP cellular interaction including uptake, intracellular fate and induction of cell response but also require very different analytical methods. Various cutting-edge techniques have emerged during the last years which can quantify the NPs based on their distinctive characteristics, for example -chemical composition, optical properties, magnetic properties or electron density. Comprehensive reviews and in-depth discussions are readily available [1][2][3][4] and are not reiterated herein. For the purpose of this commentary, we have selected some of the most widely used analytical techniques in the nanoscientific biomedical community based on a review of the existing nanotoxicology studies published within the scope of the last 5 years. Noteworthy, we do not differentiate here between intracellular and cell associated NPs which can of course also influence the choice of a method, however, it is an important point which has to be considered for the interpretation of the result. And it is important to add that, depending on the community and the research field, quanti-

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Structural Analyses of DP‐1, a Protein with the Ability To Bind Gold Nanoparticles, by Using Nuclear Magnetic Resonance Spectroscopy

Futagawa,

Tang,

Kato

et al. 2023

ChemBioChem

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Gold nanoparticles (AuNPs) consisting of metallic gold are applied in various fields owing to their characteristic physical properties. Collimonas sp. D‐25 (D‐25) is a Gram‐negative bacterium obtained from soil, compost, and other environmental materials in the Akita Prefecture. DP‐1 is a water‐soluble protein found in D‐25 that binds specifically to AuNPs and retains the nano‐sized AuNPs with high stability. This study aimed to identify the part of DP‐1 interacting with AuNPs and determine its 3D structure in solution using nuclear magnetic resonance (NMR). The peptide fragments obtained by trypsin digestion were examined for their AuNP‐binding capacity to determine the key Au‐binding domain of DP‐1. A fragment consisting of 16 amino acid residues (GHAATPEQYGVVTANK) was identified as the peptide with the highest binding activity. Structural analyses of this peptide indicated that the main chain was elongated, and negatively charged residues in the side chain were exposed on the surface by incorporating AuNPs. These results suggest that DP‐1 interacts with AuNPs via negatively charged residues and extended hydrophobic residues for protein‐protein interactions. The structural data also provide new insights into biomimetic technologies.

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Measuring and modelling cell-to-cell variation in uptake of gold nanoparticles

Cited by 40 publications

References 32 publications

Direct quantification of rare earth doped titania nanoparticles in individual human cells

Direct quantification of rare earth doped titania nanoparticles in individual human cells

Quantifying Nanoparticle Cellular Uptake: Which Method is Best?

Structural Analyses of DP‐1, a Protein with the Ability To Bind Gold Nanoparticles, by Using Nuclear Magnetic Resonance Spectroscopy

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