https://doi.org/10.1140/epje/i2009-10472-0
Regular Article
Fibronectin adsorption studied using neutron reflectometry and complementary techniques
1
Leeds Dental Institute, University of Leeds, LS2 9LU, Leeds, UK
2
Institut Laue-Langevin, 6 rue J. Horowitz, 38042, Grenoble, France
3
School of Dentistry, University of Michigan, Ann Arbor, MI, USA
* e-mail: m.al-jawad@leeds.ac.uk
Received:
12
January
2009
Accepted:
7
May
2009
Published online:
24
June
2009
In implantology it is known that fibronectin affects cell-substrate adhesion, consequently, the structure and composition of the initially adsorbed fibronectin layer to a large extent determines the biological response to a biomaterial implanted into the body. In this study we have used neutron reflectometry and quartz-crystal microbalance with dissipation to investigate the amount of fibronectin adsorbed, the layer density, thickness and structure of films adsorbed to polished silicon oxide surfaces. We have cultured MG63 osteoblast-like cells on surfaces coated and uncoated with fibronectin and monitored the cellular response to these surfaces. The results show that at fibronectin concentrations in the range 0.01 to 0.1mg/ml a single highly hydrated layer of fibronectin approximately 40-50Å in thickness adsorbs to a polished silicon oxide surface and is likely to correspond to one diffuse monolayer of fibronectin arranged side-on. Cells cultured on this fibronectin layer have dramatically different morphology and growth to those grown on bare surfaces. Using a model silicon oxide surface has enabled us to study the substrate/protein interface, together with the impact of a fibronectin layer on the cellular response using consistent experimental conditions across a unique set of experimental techniques.
PACS: 61.05.fj Neutron reflectometry – / 68.08.-p Liquid-solid interfaces – / 68.43.Mn Adsorption kinetics –
© EDP Sciences, SIF, Springer-Verlag Berlin Heidelberg, 2009