Translocation mechanisms of chemically functionalised carbon nanotubes across plasma membranes

Lacerda, Lara; Russier, Julie; Pastorin, Giorgia; Herrero, M. Antonia; Venturelli, Enrica; Dumortier, Hélène; Al‐Jamal, Khuloud T.; Prato, Maurizio; Kostarelos, Kostas; Bianco, Alberto

doi:10.1016/j.biomaterials.2012.01.024

Cited by 226 publications

(178 citation statements)

References 34 publications

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“…Subsequently, the different GFP variants (1 mM) were added to the supernatant containing the inhibitors and the cells were incubated at 37 C for 2 h. The cell uptake was then quantified using fluorescence as mentioned above. Under the same inhibition conditions, cell uptake of endocytosis markers (Alex-488 transferrin as clathrin-mediated endocytosis marker [33], BODIPYelactosylceramide as caveolaemediated endocytosis marker [34], FITCedextran as macropinocytosis marker [31]) were inhibited by >80%. The effect of temperature block was studied by preincubating the cells at 4 C for 30 min and treatment with GFP variants (1 mM) for 2 h at 4 C.…”

Section: Exploring Uptake Pathways Using Different Endocytosis Inhibimentioning

confidence: 95%

Enhancing cellular uptake of GFP via unfolded supercharged protein tags

Pesce

Kolbe

et al. 2013

Biomaterials

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a b s t r a c tOne of the barriers to the development of protein therapeutics is effective delivery to mammalian cells. The proteins must maintain a careful balance of polar moieties to enable administration and distribution and hydrophobic character to minimize cell toxicity. Numerous strategies have been applied to this end, from appending additional cationic peptides to supercharging the protein itself, sometimes with limited success. Here we present a strategy that combines these methods, by equipping a protein with supercharged elastin-like polypeptide (ELP) tags. We monitored cellular uptake and cell viability for GFP reporter proteins outfitted with a range of ELP tags and demonstrated enhanced uptake that correlates with the number of positive charges, while maintaining remarkably low cytotoxicity and resistance to degradation in the cell. GFP uptake proceeded mainly through caveolae-mediated endocytosis and we observed GFP emission inside the cells over extended time (up to 48 h). Low toxicity combined with high molecular weights of the tag opens the way to simultaneously optimize cell uptake and pharmacokinetic parameters. Thus, cationic supercharged ELP tags show great potential to improve the therapeutic profile of protein drugs leading to more efficient and safer biotherapeutics.

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Section: Exploring Uptake Pathways Using Different Endocytosis Inhibimentioning

confidence: 95%

Enhancing cellular uptake of GFP via unfolded supercharged protein tags

Pesce

Kolbe

et al. 2013

Biomaterials

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“…Recent reports on particle translocation also supported the mechanism of passive diffusion of particles into the cell membranes. 29,30 In this study, Fe 3 O 4 @C particles were observed in aggregation but not in vesicular formation; thus, they would be internalized by an energy-independent diffusion process. Further study is needed to confirm the hypothesis that the graphitic surface of the particles could cause passive diffusion of the particles into cells.…”

Section: Cytotoxicity and Cellular Uptake Of Fe 3 O 4 @Cmentioning

confidence: 58%

Photothermal cancer therapy using graphitic carbon–coated magnetic particles prepared by one-pot synthesis

Lee¹,

Sanetuntikul²,

Choi³

et al. 2014

IJN

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We describe here a simple synthetic strategy for the fabrication of carbon-coated Fe 3 O 4 (Fe 3 O 4 @C) particles using a single-component precursor, iron (III) diethylenetriaminepentaacetic acid complex. Physicochemical analyses revealed that the core of the synthesized particles consists of ferromagnetic Fe 3 O 4 material ranging several hundred nanometers, embedded in nitrogen-doped graphitic carbon with a thickness of ~120 nm. Because of their photothermal activity (absorption of near-infrared [NIR] light), the Fe 3 O 4 @C particles have been investigated for photothermal therapeutic applications. An example of one such application would be the use of Fe 3 O 4 @C particles in human adenocarcinoma A549 cells by means of NIR-triggered cell death. In this system, the Fe 3 O 4 @C can rapidly generate heat, causing >98% cell death within 10 minutes under 808 nm NIR laser irradiation (2.3 W cm −2 ). These Fe 3 O 4 @C particles provided a superior photothermal therapeutic effect by intratumoral delivery and NIR irradiation of tumor xenografts. These results demonstrate that one-pot synthesis of carbon-coated magnetic particles could provide promising materials for future clinical applications and encourage further investigation of this simple method.

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“…In our previous study, we found evidence that at least 30%-50% of chemically functionalized MWCNTs were taken up by cells through an energy-independent passive mechanism. 49 This characteristic makes nanotubes loaded with therapeutic or diagnostic cargos extremely interesting, as the release of active molecules directly into the cytoplasm increases their biological activity and therapeutic efficacy. The results obtained from this study indicated that the FITCfunctionalized MWCNTs were able to penetrate the cEND monolayers, reaching full equilibration in the Transwell ® system after 48 hours.…”

Section: Resultsmentioning

confidence: 99%

Blood–brain barrier transport studies, aggregation, and molecular dynamics simulation of multiwalled carbon nanotube functionalized with fluorescein isothiocyanate

Shityakov¹,

Salvador²,

Pastorin³

et al. 2015

IJN

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In this study, the ability of a multiwalled carbon nanotube functionalized with fluorescein isothiocyanate (MWCNT–FITC) was assessed as a prospective central nervous system-targeting drug delivery system to permeate the blood–brain barrier. The results indicated that the MWCNT–FITC conjugate is able to penetrate microvascular cerebral endothelial monolayers; its concentrations in the Transwell ® system were fully equilibrated after 48 hours. Cell viability test, together with phase-contrast and fluorescence microscopies, did not detect any signs of MWCNT–FITC toxicity on the cerebral endothelial cells. These microscopic techniques also revealed presumably the intracellular localization of fluorescent MWCNT–FITCs apart from their massive nonfluorescent accumulation on the cellular surface due to nanotube lipophilic properties. In addition, the 1,000 ps molecular dynamics simulation in vacuo discovered the phenomenon of carbon nanotube aggregation driven by van der Waals forces via MWCNT–FITC rapid dissociation as an intermediate phase.

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Translocation mechanisms of chemically functionalised carbon nanotubes across plasma membranes

Cited by 226 publications

References 34 publications

Enhancing cellular uptake of GFP via unfolded supercharged protein tags

Enhancing cellular uptake of GFP via unfolded supercharged protein tags

Photothermal cancer therapy using graphitic carbon–coated magnetic particles prepared by one-pot synthesis

Blood–brain barrier transport studies, aggregation, and molecular dynamics simulation of multiwalled carbon nanotube functionalized with fluorescein isothiocyanate

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Translocation mechanisms of chemically functionalised carbon nanotubes across plasma membranes

Cited by 226 publications

References 34 publications

Enhancing cellular uptake of GFP via unfolded supercharged protein tags

Enhancing cellular uptake of GFP via unfolded supercharged protein tags

Photothermal cancer therapy using graphitic carbon&ndash;coated magnetic particles prepared by&nbsp;one-pot synthesis

Blood&ndash;brain barrier transport studies, aggregation, and molecular dynamics simulation of multiwalled carbon nanotube functionalized with fluorescein isothiocyanate

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Product

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Photothermal cancer therapy using graphitic carbon–coated magnetic particles prepared by one-pot synthesis

Blood–brain barrier transport studies, aggregation, and molecular dynamics simulation of multiwalled carbon nanotube functionalized with fluorescein isothiocyanate