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Quantification of intracellular payload release from polymersome nanoparticles

Quantification of intracellular payload release from polymersome nanoparticles
Quantification of intracellular payload release from polymersome nanoparticles

Polymersome nanoparticles (PMs) are attractive candidates for spatio-temporal controlled delivery of therapeutic agents. Although many studies have addressed cellular uptake of solid nanoparticles, there is very little data available on intracellular release of molecules encapsulated in membranous carriers, such as polymersomes. Here, we addressed this by developing a quantitative assay based on the hydrophilic dye, fluorescein. Fluorescein was encapsulated stably in PMs of mean diameter 85 nm, with minimal leakage after sustained dialysis. No fluorescence was detectable from fluorescein PMs, indicating quenching. Following incubation of L929 cells with fluorescein PMs, there was a gradual increase in intracellular fluorescence, indicating PM disruption and cytosolic release of fluorescein. By combining absorbance measurements with flow cytometry, we quantified the real-time intracellular release of a fluorescein at a single-cell resolution. We found that 173 ± 38 polymersomes released their payload per cell, with significant heterogeneity in uptake, despite controlled synchronisation of cell cycle. This novel method for quantification of the release of compounds from nanoparticles provides fundamental information on cellular uptake of nanoparticle-encapsulated compounds. It also illustrates the stochastic nature of population distribution in homogeneous cell populations, a factor that must be taken into account in clinical use of this technology.

29460
Scarpa, Edoardo
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Bailey, Joanne L.
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Janeczek, Agnieszka A.
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Stumpf, Patrick S.
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Johnston, Alexander H.
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Oreffo, Richard O.C.
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Woo, Yin L.
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Cheong, Ying C.
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Evans, Nicholas D.
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Newman, Tracey A.
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Scarpa, Edoardo
e7d094b2-e64f-4b4c-a452-d478a912b171
Bailey, Joanne L.
88d5f9d3-4f7b-473d-8bbb-2d69fda74914
Janeczek, Agnieszka A.
71e60ef8-107b-44bd-9c0c-18d5e534ff21
Stumpf, Patrick S.
dfdb286c-b321-46d3-8ba2-85b3b4a7f092
Johnston, Alexander H.
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Oreffo, Richard O.C.
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Woo, Yin L.
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Cheong, Ying C.
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Evans, Nicholas D.
06a05c97-bfed-4abb-9244-34ec9f4b4b95
Newman, Tracey A.
322290cb-2e9c-445d-a047-00b1bea39a25

Scarpa, Edoardo, Bailey, Joanne L., Janeczek, Agnieszka A., Stumpf, Patrick S., Johnston, Alexander H., Oreffo, Richard O.C., Woo, Yin L., Cheong, Ying C., Evans, Nicholas D. and Newman, Tracey A. (2016) Quantification of intracellular payload release from polymersome nanoparticles. Scientific Reports, 6 (29460), 29460. (doi:10.1038/srep29460).

Record type: Article

Abstract

Polymersome nanoparticles (PMs) are attractive candidates for spatio-temporal controlled delivery of therapeutic agents. Although many studies have addressed cellular uptake of solid nanoparticles, there is very little data available on intracellular release of molecules encapsulated in membranous carriers, such as polymersomes. Here, we addressed this by developing a quantitative assay based on the hydrophilic dye, fluorescein. Fluorescein was encapsulated stably in PMs of mean diameter 85 nm, with minimal leakage after sustained dialysis. No fluorescence was detectable from fluorescein PMs, indicating quenching. Following incubation of L929 cells with fluorescein PMs, there was a gradual increase in intracellular fluorescence, indicating PM disruption and cytosolic release of fluorescein. By combining absorbance measurements with flow cytometry, we quantified the real-time intracellular release of a fluorescein at a single-cell resolution. We found that 173 ± 38 polymersomes released their payload per cell, with significant heterogeneity in uptake, despite controlled synchronisation of cell cycle. This novel method for quantification of the release of compounds from nanoparticles provides fundamental information on cellular uptake of nanoparticle-encapsulated compounds. It also illustrates the stochastic nature of population distribution in homogeneous cell populations, a factor that must be taken into account in clinical use of this technology.

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SREP-fluorescein paper.pdf - Accepted Manuscript
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More information

Accepted/In Press date: 13 June 2016
e-pub ahead of print date: 11 July 2016
Published date: 11 July 2016
Organisations: Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 396821
URI: https://eprints.soton.ac.uk/id/eprint/396821
PURE UUID: 89aa08e8-8a8a-41b9-814a-e34eea538a24
ORCID for Agnieszka A. Janeczek: ORCID iD orcid.org/0000-0002-9745-8346
ORCID for Patrick S. Stumpf: ORCID iD orcid.org/0000-0003-0862-0290
ORCID for Richard O.C. Oreffo: ORCID iD orcid.org/0000-0001-5995-6726
ORCID for Ying C. Cheong: ORCID iD orcid.org/0000-0001-7687-4597
ORCID for Tracey A. Newman: ORCID iD orcid.org/0000-0002-3727-9258

Catalogue record

Date deposited: 25 Jul 2016 15:34
Last modified: 03 Dec 2019 06:40

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Contributors

Author: Edoardo Scarpa
Author: Joanne L. Bailey
Author: Agnieszka A. Janeczek ORCID iD
Author: Patrick S. Stumpf ORCID iD
Author: Alexander H. Johnston
Author: Yin L. Woo
Author: Ying C. Cheong ORCID iD

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