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Real-Time Activity Bioassay of Single Osteoclasts Using a Silicon Nanocrystal-Impregnated Artificial Matrix

Lookup NU author(s): Naif Alsharif, Dr Mark Birch, Dr Ben Horrocks, Dr Harish Datta

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Abstract

There is a lack of suitable in vitro techniques for assaying osteoclastic (OC) resorptive activity in real-time; the existing assays are cumbersome, insensitive and retrospective. We have developed a sensitive real-time assay by impregnating a hydroxyapatite matrix with alkyl-capped silicon nanocrystals. Resorption of the matrix by OC releases the nanocrystals, which are internalised by the cell and detected as an increase in OC luminescence. Our choice of these nanocrystals was motivated by their pH-independent luminescence, which is proportional to concentration over a wide range, and by their rapid uptake without acute cytotoxicity. In this in vitro assay, OCs are inhibited by calcitonin (CT), methyl-b-cyclodextrin (MCD) and filipin, and stimulated by RANKL in the expected manner. The kinetics of the assay exhibit a lag phase followed by a growth of cell luminescence intensity and the whole time-course is satisfactorily described by the logistic equation.


Publication metadata

Author(s): Alsharif NH, Al-Said SAF, Birch MA, Horrocks BR, Datta HK

Publication type: Article

Publication status: Published

Journal: Small

Year: 2013

Volume: 9

Issue: 21

Pages: 3685-3692

Print publication date: 06/05/2013

Date deposited: 28/03/2013

ISSN (print): 1613-6810

ISSN (electronic): 1613-6829

Publisher: Wiley - VCH Verlag GmbH & Co. KGaA

URL: http://dx.doi.org/10.1002/smll.201203184

DOI: 10.1002/smll.201203184


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