Covalent immobilisation of tropoelastin on a plasma deposited interface for enhancement of endothelialisation on metal surfaces

Stability of a Therapeutic Layer of Immobilized Recombinant Human Tropoelastin on a Plasma-Activated Coated Surface

“…This binding has been shown to resist removal by stringent SDS washing, implying a covalent interaction (13), important for the application of a stable protein coating. The PAC surface is durable and biocompatible (14,15), while covalently immobilized TE increases endothelial cell attachment and proliferation (13). Additionally, the PAC surface and TE-coated PAC exhibit reduced thrombogenicity compared to 316L SS (14), supporting the use of this therapeutic protein coating technology to facilitate the bio-integration of blood-contacting medical devices.…”

“…The PAC is a surface composed of carbon and nitrogen deposited through a plasma process, which allows binding of proteins through well-characterized free radical species (12). This binding has been shown to resist removal by stringent SDS washing, implying a covalent interaction (13), important for the application of a stable protein coating. The PAC surface is durable and biocompatible (14,15), while covalently immobilized TE increases endothelial cell attachment and proliferation (13).…”

“…Although soluble TE is particularly cleaved by human plasma kallikrein (8), the degradation of covalently immobilized TE has not been explored. It might reasonably be expected that a monolayer of immobilized protein would resist proteolysis through steric hindrance to proteases due to the proximity of a modified PAC surface (13,14,16).…”

Free radical functionalization of surfaces to prevent adverse responses to biomedical devices

“…An effective method of creating buried radicals is to treat an organic polymer with energetic ions (13,14). After treatment with energetic ions, either postformation or during their deposition, these surfaces strongly immobilize proteins (15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28)(29) and provide a means of cloaking biomaterial surfaces. What is required is a means of controlling the density of radicals to bind a full protein monolayer that is not compromised by excessive numbers of covalent bonds, while giving sufficient shelf life of the binding property for practical applications.…”

Elastin Based Constructs

“…rhTE exhibits many properties of native tropoelastin including the ability to coacervate under physiological conditions and be cross-linked in vitro to form insoluble elastin fibers (Vrhovski, Jensen et al 1997;Muiznieks, Jensen et al 2003). rhTE promotes endothelial cell and fibroblast attachment, spreading and proliferation when used as a surface coating (Bax, Rodgers et al 2009;Rnjak, Li et al 2009;Wise, Byrom et al 2011) and improves the biocompatibility of implanted devices (Yin, Wise et al 2009;Wilson, Gibson et al 2010). Representative photographs of a synthetic human elastin scaffold (A) before cross-linking and (B-C) after cross-linking with hexamethylene diisocyanate and wetting with phosphate buffered saline.…”