The University of Southampton
University of Southampton Institutional Repository
Warning ePrints Soton is experiencing an issue with some file downloads not being available. We are working hard to fix this. Please bear with us.

C60 fullerene localization and membrane interactions in RAW 264.7 immortalized mouse macrophages

C60 fullerene localization and membrane interactions in RAW 264.7 immortalized mouse macrophages
C60 fullerene localization and membrane interactions in RAW 264.7 immortalized mouse macrophages
There continues to be a significant increase in the number and complexity of hydrophobic nanomaterials that are engineered for a variety of commercial purposes making human exposure a significant health concern. This study uses a combination of biophysical, biochemical and computational methods to probe potential mechanisms for uptake of C60 nanoparticles into various compartments of living immune cells. Cultures of RAW 264.7 immortalized murine macrophage were used as a canonical model of immune-competent cells that are likely to provide the first line of defense following inhalation. Modes of entry studied were endocytosis/pinocytosis and passive permeation of cellular membranes. The evidence suggests marginal uptake of C60 clusters is achieved through endocytosis/pinocytosis, and that passive diffusion into membranes provides a significant source of biologically-available nanomaterial. Computational modeling of both a single molecule and a small cluster of fullerenes predicts that low concentrations of fullerenes enter the membrane individually and produce limited perturbation; however, at higher concentrations the clusters in the membrane causes deformation of the membrane. These findings are bolstered by nuclear magnetic resonance (NMR) of model membranes that reveal deformation of the cell membrane upon exposure to high concentrations of fullerenes. The atomistic and NMR models fail to explain escape of the particle out of biological membranes, but are limited to idealized systems that do not completely recapitulate the complexity of cell membranes. The surprising contribution of passive modes of cellular entry provides new avenues for toxicological research that go beyond the pharmacological inhibition of bulk transport systems such as pinocytosis.
2040-3364
4134-4144
Russ, K.A.
76de9de1-f1c0-4086-9b4d-06bc11212e51
Elvati, P.
40a22547-9c48-4aa1-ad2b-5dc8f37b44ca
Parsonage, Tina, Louise
652160d8-72de-4846-a644-e9de62276706
Dews, A.
4319d2bf-d27e-443b-ae14-cb0a39f0f9dd
Jarvis, J.A.
d88eea9b-525e-40f9-ab4a-98d12b7fe7ac
Ray, M.
7f2001c1-5d01-4a81-b238-0a70f6e176e8
Schneider, B.
60e6080e-a99b-4712-b758-b3b50ba0e367
Smith, P.J.S.
003de469-9420-4f12-8f0e-8e8d76d28d6c
Williamson, P.T.F.
0b7715c6-b60e-4e95-a1b1-6afc8b9f372a
Violi, A.
00275f8f-0719-4150-9647-d896d36c3c27
Philbert, M.A.
6f0046de-c23d-41d5-9a24-7107e912dd8c
Russ, K.A.
76de9de1-f1c0-4086-9b4d-06bc11212e51
Elvati, P.
40a22547-9c48-4aa1-ad2b-5dc8f37b44ca
Parsonage, Tina, Louise
652160d8-72de-4846-a644-e9de62276706
Dews, A.
4319d2bf-d27e-443b-ae14-cb0a39f0f9dd
Jarvis, J.A.
d88eea9b-525e-40f9-ab4a-98d12b7fe7ac
Ray, M.
7f2001c1-5d01-4a81-b238-0a70f6e176e8
Schneider, B.
60e6080e-a99b-4712-b758-b3b50ba0e367
Smith, P.J.S.
003de469-9420-4f12-8f0e-8e8d76d28d6c
Williamson, P.T.F.
0b7715c6-b60e-4e95-a1b1-6afc8b9f372a
Violi, A.
00275f8f-0719-4150-9647-d896d36c3c27
Philbert, M.A.
6f0046de-c23d-41d5-9a24-7107e912dd8c

Russ, K.A., Elvati, P., Parsonage, Tina, Louise, Dews, A., Jarvis, J.A., Ray, M., Schneider, B., Smith, P.J.S., Williamson, P.T.F., Violi, A. and Philbert, M.A. (2016) C60 fullerene localization and membrane interactions in RAW 264.7 immortalized mouse macrophages. Nanoscale, 8 (7), 4134-4144. (doi:10.1039/c5nr07003a).

Record type: Article

Abstract

There continues to be a significant increase in the number and complexity of hydrophobic nanomaterials that are engineered for a variety of commercial purposes making human exposure a significant health concern. This study uses a combination of biophysical, biochemical and computational methods to probe potential mechanisms for uptake of C60 nanoparticles into various compartments of living immune cells. Cultures of RAW 264.7 immortalized murine macrophage were used as a canonical model of immune-competent cells that are likely to provide the first line of defense following inhalation. Modes of entry studied were endocytosis/pinocytosis and passive permeation of cellular membranes. The evidence suggests marginal uptake of C60 clusters is achieved through endocytosis/pinocytosis, and that passive diffusion into membranes provides a significant source of biologically-available nanomaterial. Computational modeling of both a single molecule and a small cluster of fullerenes predicts that low concentrations of fullerenes enter the membrane individually and produce limited perturbation; however, at higher concentrations the clusters in the membrane causes deformation of the membrane. These findings are bolstered by nuclear magnetic resonance (NMR) of model membranes that reveal deformation of the cell membrane upon exposure to high concentrations of fullerenes. The atomistic and NMR models fail to explain escape of the particle out of biological membranes, but are limited to idealized systems that do not completely recapitulate the complexity of cell membranes. The surprising contribution of passive modes of cellular entry provides new avenues for toxicological research that go beyond the pharmacological inhibition of bulk transport systems such as pinocytosis.

Text
Russ2016.pdf - Accepted Manuscript
Download (2MB)

More information

Accepted/In Press date: 21 January 2016
e-pub ahead of print date: 25 January 2016
Published date: 2016
Organisations: Faculty of Natural and Environmental Sciences, Optoelectronics Research Centre

Identifiers

Local EPrints ID: 387905
URI: http://eprints.soton.ac.uk/id/eprint/387905
ISSN: 2040-3364
PURE UUID: e64649b5-31ef-4e16-b5e1-f3c6cca98b44
ORCID for P.J.S. Smith: ORCID iD orcid.org/0000-0003-4400-6853
ORCID for P.T.F. Williamson: ORCID iD orcid.org/0000-0002-0231-8640

Catalogue record

Date deposited: 17 Feb 2016 11:33
Last modified: 21 Nov 2021 03:04

Export record

Altmetrics

Contributors

Author: K.A. Russ
Author: P. Elvati
Author: Tina, Louise Parsonage
Author: A. Dews
Author: J.A. Jarvis
Author: M. Ray
Author: B. Schneider
Author: P.J.S. Smith ORCID iD
Author: A. Violi
Author: M.A. Philbert

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of http://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×