Modification of material surfaces aimed at bestowing them with antimicrobial properties is a promising approach in the development of new biomaterials. Antimicrobial peptides (AMPs) are an attractive alternative to conventional antibiotics because of lack of toxicity, inherently high selectivity, and absence of immune response. As the antimicrobial mode of action of the AMP cathelin LL37 is formation of pores and disruption of microbial membrane, the purpose of the present study was to develop and test a method of covalent immobilization of LL37 on titanium surface. The application of a flexible hydrophilic poly(ethylene glycol) spacer and selective N-terminal conjugation of LL37 resulted in a surface peptide layer which was capable of killing bacteria on contact.
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 land c-RGD peptides) on EC cell coverage and proliferation was evaluated. After 24 h and after 3 d, surface coverage of adherent cells was quantifi ed and an alamarBlue® proliferation assay was conducted. After 24 h, l-RGD modifi ed surfaces showed a signifi cantly 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 signifi cant. After 3 d, c-RGD surfaces showed a signifi cantly higher cell coverage than untreated Ti, silanised and l-RGD titanium surfaces (all p<0.0001). After 24 h, c-RGD modified surfaces showed signifi cant 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 signifi cantly untreated titanium (p=0.004), silanised (p=0.001) and l-RGD surfaces (p=0.023), whereas no signifi cant 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 modifi cations of titanium surfaces with c-RGD are a promising approach in promoting endothelial cell growth.
Direct surface modification of biodegradable polycaprolactone (PCL) was performed without the necessity of synthesis of functionisable co-polymers. An easy-to-perform three-step procedure consisting of amination, reaction with hetero-bifunctional cross-linkers and conjugation of an RGD-motif-containing peptide was used to modify polymer films and improve the attachment of endothelial cells. The biological activity of modified surfaces was assessed by estimating microvascular endothelial cell attachment. Covalent coating with RGD resulted in an approximately 11-fold increase of endothelial cell attachment on modified PCL surfaces compared with untreated polymer. The specificity of the attachment enhancement was confirmed by using a control peptide. It is concluded that chemical surface modification is an appropriate method of rendering degradable polymers, such as PCL, cell-adhesive.
Polytetrafluoroethylene (PTFE), a frequently utilized polymer for the fabrication of synthetic vascular grafts, was surface-modified by means of a wet-chemical process. The inherently non-cell-adhesive polymer does not support cellular attachment, a prerequisite for the endothelialization of luminal surface grafts in small diameter applications. To impart the material with cell-adhesive properties a treatment with sodium-naphthalene provided a basis for the subsequent immobilization of the adhesion promoting RGD-peptide using a hydroxy- and amine-reactive crosslinker. Successful conjugation was shown with cell culture experiments which demonstrated excellent endothelial cell growth on the modified surfaces.
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