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The development and in vitro characterisation of an intracellular nanosensor responsive to reactive oxygen species

Lookup NU author(s): Dr James Henderson, Dr David Fulton, Emeritus Professor Calum McNeil, Dr Philip Manning

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Abstract

Advances in sensor technologies have enhanced our understanding of the roles played by reactive oxygen species (ROS) in a number of physiological and pathological processes. However, high inter-reactivity and short life spans has made real-time monitoring of ROS in cellular systems challenging. Fluorescent dyes capable of intracellular ROS measurements have been reported. However, these dyes are known to be intrinsically cytotoxic and thus can potentially significantly alter cellular metabolism and adversely influence in vitro data. Reported here is the development and in vitro application of a novel ROS responsive nanosensor, based on PEBBLE (Probes Encapsulated By Biologically Localised Embedding) technology. The ROS sensitive fluorescent probe dihydrorhodamine 123 (DHR 123) was employed as the sensing element of the PEBBLE through entrapment within a porous, bio-inert polyacrylamide nanostructure enabling passive monitoring of free radical flux within the intracellular environment. Successful delivery of the nanosensors into NR8383 rat alveolar macrophage cells via phagocytosis was achieved. Stimulation of PEBBLE loaded NR8383 cells with phorbol-12-myristate-13-acetate (PMA) enabled real time monitoring of ROS generation within the cell without affecting cellular viability. These data suggest that PEBBLE nanosensors could offer significant advantages over existing technologies used in monitoring the intracellular environment. (C) 2009 Elsevier B.V. All rights reserved.


Publication metadata

Author(s): Henderson JR, Fulton DA, McNeil CJ, Manning P

Publication type: Article

Publication status: Published

Journal: Biosensors and Bioelectronics

Year: 2009

Volume: 24

Issue: 12

Pages: 3608-3614

ISSN (print): 0956-5663

ISSN (electronic): 1873-4235

Publisher: Elsevier BV

URL: http://dx.doi.org/10.1016/j.bios.2009.05.029

DOI: 10.1016/j.bios.2009.05.029


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