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Adhesion formation of primary human osteoblasts and the functional response of mesenchymal stem cells to 330nm deep microgrooves

Adhesion formation of primary human osteoblasts and the functional response of mesenchymal stem cells to 330nm deep microgrooves
Adhesion formation of primary human osteoblasts and the functional response of mesenchymal stem cells to 330nm deep microgrooves
The surface microtexture of an orthopaedic device can regulate cellular adhesion, a process fundamental in the initiation of osteoinduction and osteogenesis. Advances in fabrication techniques have evolved to include the field of surface modification; in particular, nanotechnology has allowed for the development of experimental nanoscale substrates for investigation into cell nanofeature interactions. Here primary human osteoblasts (HOBs) were cultured on ordered nanoscale groove/ridge arrays fabricated by photolithography. Grooves were 330nm deep and either 10, 25 or 100mum in width. Adhesion subtypes in HOBs were quantified by immunofluorescent microscopy and cell-substrate interactions were investigated via immunocytochemistry with scanning electron microscopy. To further investigate the effects of these substrates on cellular function, 1.7K gene microarray analysis was used to establish gene regulation profiles of mesenchymal stem cells cultured on these nanotopographies. Nanotopographies significantly affected the formation of focal complexes (FXs), focal adhesions (FAs) and supermature adhesions (SMAs). Planar control substrates induced widespread adhesion formation; 100mum wide groove/ridge arrays did not significantly affect adhesion formation yet induced upregulation of genes involved in skeletal development and increased osteospecific function; 25mum wide groove/ridge arrays were associated with a reduction in SMA and an increase in FX formation; and 10mum wide groove/ridge arrays significantly reduced osteoblast adhesion and induced an interplay of up- and downregulation of gene expression. This study indicates that groove/ridge topographies are important modulators of both cellular adhesion and osteospecific function and, critically, that groove/ridge width is important in determining cellular response.
development, function, nanotechnology, human, analysis, research, expression, gene, osteogenesis, gene expression, gene-expression, mesenchymal stem cells, stem cells, osteoblasts
1742-5689
1231-1242
Biggs, M.J.P.
8755a848-e5e0-430d-88fb-0c7fc9bb0573
Richards, R.G.
fae1269a-4287-4988-8847-7481edcbf1bb
McFarlane, S.
237f6e92-2a4c-4bc1-ae22-f1856417d1cd
Wilkinson, C.D.W.
44d22b13-4961-49bc-a9ac-a1089b849d88
Oreffo, R.O.C.
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
Dalby, M.J.
b1f2e705-d463-433a-9e27-bcfe17568ddb
Biggs, M.J.P.
8755a848-e5e0-430d-88fb-0c7fc9bb0573
Richards, R.G.
fae1269a-4287-4988-8847-7481edcbf1bb
McFarlane, S.
237f6e92-2a4c-4bc1-ae22-f1856417d1cd
Wilkinson, C.D.W.
44d22b13-4961-49bc-a9ac-a1089b849d88
Oreffo, R.O.C.
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
Dalby, M.J.
b1f2e705-d463-433a-9e27-bcfe17568ddb

Biggs, M.J.P., Richards, R.G., McFarlane, S., Wilkinson, C.D.W., Oreffo, R.O.C. and Dalby, M.J. (2008) Adhesion formation of primary human osteoblasts and the functional response of mesenchymal stem cells to 330nm deep microgrooves. Journal of the Royal Society Interface, 5 (27), 1231-1242. (doi:10.1098/rsif.2008.0035).

Record type: Article

Abstract

The surface microtexture of an orthopaedic device can regulate cellular adhesion, a process fundamental in the initiation of osteoinduction and osteogenesis. Advances in fabrication techniques have evolved to include the field of surface modification; in particular, nanotechnology has allowed for the development of experimental nanoscale substrates for investigation into cell nanofeature interactions. Here primary human osteoblasts (HOBs) were cultured on ordered nanoscale groove/ridge arrays fabricated by photolithography. Grooves were 330nm deep and either 10, 25 or 100mum in width. Adhesion subtypes in HOBs were quantified by immunofluorescent microscopy and cell-substrate interactions were investigated via immunocytochemistry with scanning electron microscopy. To further investigate the effects of these substrates on cellular function, 1.7K gene microarray analysis was used to establish gene regulation profiles of mesenchymal stem cells cultured on these nanotopographies. Nanotopographies significantly affected the formation of focal complexes (FXs), focal adhesions (FAs) and supermature adhesions (SMAs). Planar control substrates induced widespread adhesion formation; 100mum wide groove/ridge arrays did not significantly affect adhesion formation yet induced upregulation of genes involved in skeletal development and increased osteospecific function; 25mum wide groove/ridge arrays were associated with a reduction in SMA and an increase in FX formation; and 10mum wide groove/ridge arrays significantly reduced osteoblast adhesion and induced an interplay of up- and downregulation of gene expression. This study indicates that groove/ridge topographies are important modulators of both cellular adhesion and osteospecific function and, critically, that groove/ridge width is important in determining cellular response.

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More information

Published date: 6 October 2008
Keywords: development, function, nanotechnology, human, analysis, research, expression, gene, osteogenesis, gene expression, gene-expression, mesenchymal stem cells, stem cells, osteoblasts

Identifiers

Local EPrints ID: 60916
URI: https://eprints.soton.ac.uk/id/eprint/60916
ISSN: 1742-5689
PURE UUID: 62828c56-5623-4dec-8ad9-2d65efc6fef1
ORCID for R.O.C. Oreffo: ORCID iD orcid.org/0000-0001-5995-6726

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Date deposited: 26 Sep 2008
Last modified: 19 Nov 2019 01:52

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Contributors

Author: M.J.P. Biggs
Author: R.G. Richards
Author: S. McFarlane
Author: C.D.W. Wilkinson
Author: R.O.C. Oreffo ORCID iD
Author: M.J. Dalby

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