Photoluminescent carbon nanotubes interrogate the permeability of multicellular tumor spheroids.

Nanomaterials have been extensively investigated for cancer drug delivery and imaging applications. Nanoparticles that show promise in two-dimensional cell culture systems often fail in more complex environments, possibly due to the lack of penetration in dense, three-dimensional structures. Multicellular tumor spheroids are an emerging model system to investigate interactions of nanoparticles with 3D in vitro cell culture environments. Using the intrinsic near-infrared emission of semiconducting carbon nanotubes to optically reconstruct their localization within a three-dimensional volume, we resolved the relative permeability of two different multicellular tumor spheroids. Nanotube photoluminescence revealed that nanotubes rapidly internalized into MCF-7 breast cancer cell-derived spheroids, whereas they exhibited little penetration into spheroids derived from SK-136, a cell line that we developed from murine liver cancer. Characterization of the spheroids by electron microscopy and immunohistochemistry revealed large differences in the extracellular matrix and interstitial spacing, which correlated directly with nanotube penetration. This platform portends a new approach to characterize the permeability of living multicellular environments.

[1]  S. Iijima.  Helical microtubules of graphitic carbon , 1991, Nature.

[2]  Michael J. Bronikowski,et al.  Gas-phase production of carbon single-walled nanotubes from carbon monoxide via the HiPco process: A parametric study , 2001 .

[3]  L. Kunz-Schughart,et al.  Multicellular tumor spheroids: an underestimated tool is catching up again. , 2010, Journal of biotechnology.

[4]  R. Smalley,et al.  Structure-Assigned Optical Spectra of Single-Walled Carbon Nanotubes , 2002, Science.

[5]  L. Li,et al.  Deep penetration of nanoparticulate drug delivery systems into tumors: challenges and solutions. , 2013, Current medicinal chemistry.

[6]  S. Bachilo,et al.  Analyzing absorption backgrounds in single-walled carbon nanotube spectra. , 2011, ACS nano.

[7]  L. Kunz-Schughart,et al.  Multicellular tumor spheroids: intermediates between monolayer culture and in vivo tumor , 1999, Cell biology international.

[8]  H. Dai,et al.  Carbon Nanomaterials for Biological Imaging and Nanomedicinal Therapy. , 2015, Chemical reviews.

[9]  C. M. Roth,et al.  Nanoscale drug delivery systems for enhanced drug penetration into solid tumors: current progress and opportunities. , 2012, Critical reviews in biomedical engineering.

[10]  Triantafyllos Stylianopoulos,et al.  Delivery of molecular and nanoscale medicine to tumors: transport barriers and strategies. , 2011, Annual review of chemical and biomolecular engineering.

[11]  V. C. Moore,et al.  Band Gap Fluorescence from Individual Single-Walled Carbon Nanotubes , 2002, Science.

[12]  Zhuang Liu,et al.  A route to brightly fluorescent carbon nanotubes for near-infrared imaging in mice. , 2009, Nature nanotechnology.

[13]  V. Muzykantov,et al.  Multifunctional Nanoparticles: Cost Versus Benefit of Adding Targeting and Imaging Capabilities , 2012, Science.

[14]  Lijun Wu,et al.  Optical Spectroscopy of Individual Single-Walled Carbon Nanotubes of Defined Chiral Structure , 2006, Science.

[15]  Suzie H Pun,et al.  Spatio‐temporal modeling of nanoparticle delivery to multicellular tumor spheroids , 2008, Biotechnology and bioengineering.

[16]  M. Prato,et al.  How do functionalized carbon nanotubes land on, bind to and pierce through model and plasma membranes. , 2013, Nanoscale.

[17]  Thomas E. Eurell,et al.  Single‐Walled Carbon Nanotube Spectroscopy in Live Cells: Towards Long‐Term Labels and Optical Sensors , 2005 .

[18]  R. Steiner,et al.  Spatially resolved absolute diffuse reflectance measurements for noninvasive determination of the optical scattering and absorption coefficients of biological tissue. , 1996, Applied optics.

[19]  Daniel A. Heller,et al.  Treating metastatic cancer with nanotechnology , 2011, Nature Reviews Cancer.

[20]  V. Venditto,et al.  Cancer nanomedicines: so many papers and so few drugs! , 2013, Advanced drug delivery reviews.

[21]  M. J. Loureiro,et al.  Pharmaceutical characterization of solid and dispersed carbon nanotubes as nanoexcipients , 2012, International journal of nanomedicine.

[22]  Huajian Gao,et al.  Cell entry of one-dimensional nanomaterials occurs by tip recognition and rotation. , 2011, Nature nanotechnology.

[23]  S. Bachilo,et al.  Dependence of Optical Transition Energies on Structure for Single-Walled Carbon Nanotubes in Aqueous Suspension: An Empirical Kataura Plot , 2003 .

[24]  L. weiswald,et al.  Spherical Cancer Models in Tumor Biology1 , 2015, Neoplasia.

[25]  Edward Kai-Hua Chow,et al.  Oncogene‐specific formation of chemoresistant murine hepatic cancer stem cells , 2012, Hepatology.

[26]  Johannes E. Schindelin,et al.  Fiji: an open-source platform for biological-image analysis , 2012, Nature Methods.

[27]  M. Strano,et al.  Sonication-induced changes in chiral distribution: A complication in the use of single-walled carbon nanotube fluorescence for determining species distribution , 2005 .

[28]  T Lammers,et al.  Tumour-targeted nanomedicines: principles and practice , 2008, British Journal of Cancer.

[29]  Keiran S. M. Smalley,et al.  Life ins't flat: Taking cancer biology to the next dimension , 2006, In Vitro Cellular & Developmental Biology - Animal.

[30]  B. Bauer,et al.  Length‐Dependent Uptake of DNA‐Wrapped Single‐Walled Carbon Nanotubes , 2007 .

[31]  Helen G. Hansma,et al.  Adsorption of DNA to mica, silylated mica, and minerals : characterization by atomic force microscopy , 1995 .

[32]  Brendon M. Baker,et al.  Deconstructing the third dimension – how 3D culture microenvironments alter cellular cues , 2012, Journal of Cell Science.

[33]  Ayumi Ishibashi,et al.  Individual dissolution of single-walled carbon nanotubes in aqueous solutions of steroid or sugar compounds and their Raman and near-IR spectral properties. , 2006, Chemistry.

[34]  R. Weisman,et al.  Length- and defect-dependent fluorescence efficiencies of individual single-walled carbon nanotubes. , 2012, ACS nano.

[35]  Stéphane Marcet,et al.  Hyperspectral Microscopy of Near-Infrared Fluorescence Enables 17-Chirality Carbon Nanotube Imaging , 2015, Scientific Reports.

[36]  Prakrit V. Jena,et al.  P-selectin is a nanotherapeutic delivery target in the tumor microenvironment , 2016, Science Translational Medicine.

[37]  N. Kotov,et al.  Anomalously Fast Diffusion of Targeted Carbon Nanotubes in Cellular Spheroids. , 2015, ACS nano.

[38]  P. Shaw.  Comparison of Widefield/Deconvolution and Confocal Microscopy for Three-Dimensional Imaging , 2006 .

[39]  Shuo Diao,et al.  Through-skull fluorescence imaging of the brain in a new near-infrared window , 2014, Nature Photonics.

[40]  Michael S. Strano,et al.  Size-dependent cellular uptake and expulsion of single-walled carbon nanotubes: single particle tracking and a generic uptake model for nanoparticles. , 2009, ACS nano.

[41]  M. Pasquali,et al.  Environmental and synthesis-dependent luminescence properties of individual single-walled carbon nanotubes. , 2009, ACS nano.