Tuning payload delivery in tumour cylindroids using gold nanoparticles

Journal name:
Nature Nanotechnology
Volume:
5,
Pages:
465–472
Year published:
DOI:
doi:10.1038/nnano.2010.58
Received
Accepted
Published online

Abstract

Nanoparticles have great potential as controllable drug delivery vehicles because of their size and modular functionality. Timing and location are important parameters when optimizing nanoparticles for delivery of chemotherapeutics. Here, we show that gold nanoparticles carrying either fluorescein or doxorubicin molecules move and localize differently in an in vitro three-dimensional model of tumour tissue, depending on whether the nanoparticles are positively or negatively charged. Fluorescence microscopy and mathematical modelling show that uptake, not diffusion, is the dominant mechanism in particle delivery. Our results indicate that positive particles may be more effective for drug delivery because they are taken up to a greater extent by proliferating cells. Negative particles, which diffuse more quickly, may perform better when delivering drugs deep into tissues. An understanding of how surface charge can control tissue penetration and drug release may overcome some of the current limitations in drug delivery.

At a glance

Figures

  1. Schematic showing the delivery of payload by gold nanoparticles.
    Figure 1: Schematic showing the delivery of payload by gold nanoparticles.

    a, Delivery of payload (green ovals) into tumour cylindroids by gold nanoparticles (gold circles). Cells containing released FITC–SH are in green. Viable cells are shown with smooth, solid boundaries, and necrotic cells have irregular, dashed boundaries. Dashed arrows indicate diffusion and cellular uptake. In cylindroids, nanoparticles are present in the medium outside the boundary of the cell mass. b, Intratumoral delivery by gold nanoparticles following extravasation from the vessel lumen (red circles). c, Mixed monolayer-protected gold nanoparticles loaded with thioalkylated FITC or doxorubicin (DOX). d, Cellular uptake and FITC–SH release by thiol-mediated replacement reactions.

  2. Fluorescence calibration and cellular uptake and release of FITC-AuNPs.
    Figure 2: Fluorescence calibration and cellular uptake and release of FITC–AuNPs.

    a, Normalized fluorescence intensities plotted against nanoparticle concentration. Dotted lines represent linear least-squares fitting results. For all components, fluorescence intensity was linearly proportional to concentration. b,c, Differential interference contrast (DIC) and confocal green fluorescent images of cells in monolayer culture incubated with cationic (b) and anionic (c) gold nanoparticles. Scale bars, 50 μm.

  3. Release of FITC-SH from AuNPs in tumour cylindroids.
    Figure 3: Release of FITC–SH from AuNPs in tumour cylindroids.

    a–f, Green fluorescence images (a,c,e) and corresponding intensity profiles (b,d,f) of cylindroids treated with nanoparticles. Fluorescence images were acquired after incubation with p-FITC–AuNP (a), n-FITC–AuNP (c) and FITC (e) for 11, 21 and 46 h. Dotted circles indicate cylindroid edges. Scale bar, 300 µm. Fluorescence intensity profiles are shown at 11, 21, 38 and 46 h (b,d). g,h, Change of average fluorescence intensities over time in the inner and outer regions of the cylindroids (indicated by grey bars in b and d) after incubation with p-FITC–AuNP (g) and n-FITC–AuNP (h). Errors are standard error of the mean (n = 3). i, Blue fluorescence image of a cylindroid incubated with OPA for 18 h. Scale bar, 200 µm. j, Green confocal fluorescence microscope image of the outer region of the cylindroid treated with p-FITC–AuNP. Scale bar, 20 µm.

  4. Doxorubicin release in cylindroids.
    Figure 4: Doxorubicin release in cylindroids.

    a–c, Red fluorescence images and the corresponding radial intensity profiles of cylindroids treated with positive DOX–AuNP (a), negative DOX–AuNP (b) and free DOX (c). Dotted circles indicate cylindroid edges. Scale bars, 200 µm. d,e, Ratio of average fluorescence intensities in the outer region to those in the inner region of the cylindroids, 30 h after treatment (d) and as a function of time (e). Errors are standard error of the mean, and significance was determined using Student's t-test (*P < 0.05; n = 3).

  5. Effect of surface charge on diffusivity through extracellular matrix material.
    Figure 5: Effect of surface charge on diffusivity through extracellular matrix material.

    a, Schematic representation of loading module filled with Matrigel. b, Green fluorescence images acquired from the bottom of the loading module 4 and 24 h after adding particles. Dotted lines at both ends indicate the Matrigel edges. Scale bar, 500 µm. c, Fluorescence intensity profiles in Matrigel along the x-axis as a function of time (symbols) and intensity profiles generated by fitting to a Fickian diffusion model (solid lines). d, Comparison of diffusion coefficients of FITC, cationic and anionic gold nanoparticles in Matrigel (*P < 0.005; n = 3).

  6. Rate constants of cellular uptake and predictions of particle and ligand distribution in tumours.
    Figure 6: Rate constants of cellular uptake and predictions of particle and ligand distribution in tumours.

    a,b, Fluorescence intensity profiles measured in fluorescence images of cylindroids (symbols) and those predicted by the mathematical model (solid lines). Concentrations were normalized by the maximum concentration of FITC–AuNP in the cylindroids. c, Independent concentration profiles of fluorophore bound to nanoparticles (FITC–AuNP) and released fluorophore ligand (FITC–SH) predicted by the computation model. d, Comparison of forward (k1) and reverse (k2) rate constants in different regions. Errors are standard error of the mean, and significance was determined using Student's t-test (*P < 0.05). e,f, Modelled concentrations of FITC–AuNPs and released FITC–SH as a function of distance from blood vessels of a hypothetical tumour. Clearance half-life of FITC–AuNPs was assumed to be 3 h. g, Average concentrations of FITC–AuNPs in the regions 10–70 µm (proliferating) and 75–120 µm (quiescent) from the blood vessel wall as a function of time. h, Average concentrations of released FITC in deep tumour regions (1.0–1.5 mm) as a function of time.

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Author information

Affiliations

  1. Department of Chemical Engineering, University of Massachusetts, Amherst, Massachusetts 01003-9303, USA

    • Byoungjin Kim,
    • Bhushan J. Toley &
    • Neil S. Forbes
  2. Department of Chemistry, University of Massachusetts, Amherst, Massachusetts 01003-9303, USA

    • Gang Han,
    • Chae-kyu Kim &
    • Vincent M. Rotello

Contributions

N.F. and V.R. conceived and designed the experiments. B.K., G.H., B.T. and C.K. performed the experiments. G.H. synthesized the FITC nanoparticles. C.K. synthesized the DOX nanoparticles. B.K. performed all cell and cylindroid experiments with FITC nanoparticles and wrote all mathematical models. B.T. performed all cylindroid and cell experiments with DOX nanoparticles. B.K., B.T. and N.F. analysed the data. V.R. contributed materials. B.K. and N.F. co-wrote the paper. All authors discussed the results and commented on the manuscript.

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The authors declare no competing financial interests.

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