Covalent immobilisation of tropoelastin on a plasma deposited interface for enhancement of endothelialisation on metal surfaces.
Currently available endovascular metallic implants such as stents exhibit suboptimal biocompatibility in that they re-endothelialise poorly leaving them susceptible to thrombosis. To improve the interaction of these implants with endothelial cells we developed a surface coating technology, enabling the covalent attachment of biomolecules to previously inert metal surfaces. Using horseradish peroxidase as a probe, we demonstrate that the polymerised surface can retain the presentation and activity of an immobilised protein. We further demonstrated the attachment of tropoelastin, an extracellular matrix protein critical to the correct arrangement and function of vasculature. Not only it is structurally important, but it plays a major role in supporting endothelial cell growth, while modulating smooth muscle cell infiltration. Tropoelastin was shown to bind to the surface in a covalent monolayer, supplemented with additional physisorbed multilayers on extended incubation. The physisorbed tropoelastin layers can be washed away in buffer or SDS while the first layer of tropoelastin remains tightly bound. The plasma coated stainless steel surface with immobilised tropoelastin was subsequently found to have improved biocompatibility by promoting endothelial cell attachment and proliferation relative to uncoated stainless steel controls. Tropoelastin coatings applied to otherwise inert substrates using this technology could thus have broad applications to a range of non-polymeric vascular devices.
Yin
Yongbai
Y
University of Sydney, NSW, Australia.
Wise
Steven G
SG
Nosworthy
Neil J
NJ
Waterhouse
Anna
A
Bax
Daniel V
DV
Youssef
Hani
H
Byrom
Michael J
MJ
Bilek
Marcela M M
MM
McKenzie
David R
DR
Weiss
Anthony S
AS
Ng
Martin K C
MK
D000268:Adhesiveness; D002448:Cell Adhesion; D049109:Cell Proliferation; D002478:Cells, Cultured; D003460:Crystallization; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005559:Formamides; D006735:Horseradish Peroxidase / Q000378:metabolism; D006801:Humans; D055767:Immobilized Proteins / Q000378:metabolism*; D008297:Male; D008670:Metals / Q000378:metabolism*; D011791:Quartz; D012031:Refractometry; D013499:Surface Properties; D013997:Time Factors; D014333:Tropoelastin / Q000378:metabolism*
D005559:Formamides; D055767:Immobilized Proteins; D008670:Metals; D014333:Tropoelastin; D011791:Quartz; C031066:formamide; D006735:Horseradish Peroxidase