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, 1-13, [29460]. (doi:10.1038/srep29460).
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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- Faculties (pre 2018 reorg) > Faculty of Medicine (pre 2018 reorg) > Human Development & Health (pre 2018 reorg)
Current Faculties > Faculty of Medicine > Human Development and Health > Human Development & Health (pre 2018 reorg)
Human Development and Health > Human Development & Health (pre 2018 reorg) - Faculties (pre 2018 reorg) > Faculty of Medicine (pre 2018 reorg) > Clinical & Experimental Sciences (pre 2018 reorg)
Current Faculties > Faculty of Medicine > Clinical and Experimental Sciences > Clinical & Experimental Sciences (pre 2018 reorg)
Clinical and Experimental Sciences > Clinical & Experimental Sciences (pre 2018 reorg) - Faculties (pre 2018 reorg) > Faculty of Engineering and the Environment (pre 2018 reorg) > Southampton Marine & Maritime Institute (pre 2018 reorg)
- Faculties (pre 2018 reorg) > Faculty of Natural and Environmental Sciences (pre 2018 reorg) > Institute for Life Sciences (pre 2018 reorg)
Current Faculties > Faculty of Environmental and Life Sciences > Institute for Life Sciences > Institute for Life Sciences (pre 2018 reorg)
Institute for Life Sciences > Institute for Life Sciences (pre 2018 reorg) - Faculties (pre 2018 reorg) > Faculty of Engineering and the Environment (pre 2018 reorg) > Mechanical Engineering (pre 2018 reorg) > Bioengineering Group (pre 2018 reorg)
Current Faculties > Faculty of Engineering and Physical Sciences > School of Engineering > Mechanical Engineering > Mechanical Engineering (pre 2018 reorg) > Bioengineering Group (pre 2018 reorg)
Mechanical Engineering > Mechanical Engineering (pre 2018 reorg) > Bioengineering Group (pre 2018 reorg)
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