Immobilisation of linear and cyclic RGD-peptides on titanium surfaces and their impact on endothelial cell adhesion and proliferation

Abstract

Functional coatings on titanium vascular stents and endosseous dental implants could probably enhance endothelial cell (EC) adhesion and activity with a shortening of the wound healing time and an increase of peri-implant angiogenesis during early bone formation. Therefore, the role of the structure of linear and cyclic cell adhesive peptides Arg-Gly-Asp (l-RGD and c-RGD) on differently pre-treated titanium (Ti) surfaces (untreated, silanised vs. functionalised with l- and c-RGD peptides) on EC cell coverage and proliferation was evaluated. After 24 h and after 3 d, surface coverage of adherent cells was quantified and an alamarBlue® proliferation assay was conducted. After 24 h, l-RGD modified surfaces showed a significantly better coverage of adhered cells than untreated titanium (p=0.01). Differences between l-RGD surfaces and silanised Ti (p=0.066) as well as between l-RGD and c-RGD surfaces (p=0.191) were not significant. After 3 d, c-RGD surfaces showed a significantly higher cell coverage than untreated Ti, silanised and l-RGD titanium surfaces (all p<0.0001). After 24 h, c-RGD modified surfaces showed significant higher cell proliferation compared to untreated Ti (p=0.003). However, there were no differences in proliferation between c-RGD and l-RGD (p=0.126) or c-RGD and silanised titanium (p=0.196). After 3 d, proliferation on c-RGD surfaces outranged significantly untreated titanium (p=0.004), silanised (p=0.001) and l-RGD surfaces (p=0.023), whereas no significant difference could be found between untreated Ti and l-RGD surfaces (p=0.54). According to these results, the biomimetic coating of c-RGD peptides on conventional titanium surfaces showed a positive effect on EC cell coverage and proliferation. We were able to show that modifications of titanium surfaces with c-RGD are a promising approach in promoting endothelial cell growth.