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Abstract: The development of new strategies for protein immobilization to control cell adhesion, growth and differentiation is of prime interest in the field of tissue engineering. Here we propose a versatile approach based on the interaction between two de novo designed peptides, Ecoil and Kcoil, for oriented immobilization of epidermal growth factor (EGF) on polyethylene terephthalate (PET) films. After amination of PET surfaces by ammonia plasma treatment, Kcoil peptides were covalently grafted in an oriented fashion using succinimidyl 6-[30-(2-pyridyldithio)-propionamido] hexanoate (LC-SPDP) linker, and the Kcoil-functionalized films were characterized by X-ray photoelectron spectroscopy (XPS). Bioactivity of Ecoil-EGF captured on Kcoil-functionalized PET via coiled-coil interactions was confirmed by EGF receptor phosphorylation analysis following A-431 cell attachment. We also demonstrated cell biological effects where tethered EGF enhanced adhesion, spreading and proliferation of human corneal epithelial cells compared to EGF that was either physically adsorbed or present in solution. Tethered EGF effects were most likely linked to the prolonged activation of both mitogen-activated protein kinase and phosphoinositidine 3-kinase pathways. Taken together, our results indicate that coiled-coil-based oriented immobilization is a powerful method to specifically tailor biomaterial surfaces for tissue engineering applications. [Copyright &y& Elsevier]

Abstract: Complications following endovascular aneurysm repair (EVAR) are related to deficient healing around the stent-graft (SG). New generations of SG with surface properties that foster vascular repair could overcome this limitation. Our goal was to evaluate the potential of a new nitrogen-rich plasma-polymerised biomaterial, designated PPE:N, as an external coating for polyethylene terephtalate (PET)- or polytetrafluoro-ethylene (PTFE)-based SGs, to promote healing around the implant. Thin PPE:N coatings were deposited on PET and PTFE films. Then, adhesion, growth, migration and resistance to apoptosis of vascular smooth muscle cells (VSMCs) and fibroblasts, as well as myofibroblast differentiation, were assessed in vitro. In another experimental group, chondroitin sulphate (CS), a newly described mediator of vascular repair, was added to normal culture medium, to search for possible additional benefit. PPE:N-coatings, especially on PET, increased and accelerated cell adhesion and growth, compared with control PET and with standard polystyrene culture plates (PCP). PPE:N was also found to increase the resistance to apoptosis in VSMC, an important finding as aneurysms are characterised by VMSC depletion caused by a pro-apoptotic phenotype. Addition of CS in solution further increased migration and resistance to apoptosis. In conclusion, PPE:N-coating and/or CS could promote vascular repair around SGs following EVAR. [Copyright &y& Elsevier]

Abstract: Little is known of the effect of material surfaces on stem cell differentiation. The present study has addressed the hypothesis that the interaction of mesenchymal stem cells (MSCs) with material surfaces modified by glow discharge plasma is a major regulator of osteogenic differentiation. We found that biaxially oriented polypropylene (BOPP) plasma treated in ammonia significantly reduced up-regulation of expression of osteogenic marker genes, such as alkaline phosphatase (ALP), bone sialoprotein (BSP) and osteocalcin (OC). In contrast, ALP expression was up-regulated when cultured on treated Nylon-6 polyamide (Ny-t) but was substantially reduced when cultured on its pristine counterpart (Ny-p) on day 3. On day 7, ALP expression was down-regulated with MSCs cultured on Ny-t although its expression level was up again on day 14. BSP was expressed weakly on day 3, but was up-regulated when cultured on Ny-t and Ny-p. Its expression reached its maximum on day 14 when cultured on a polystyrene control, while it was cyclically up-regulated on Ny-t. Similarly, there was a slight increase in OC expression when MSCs were cultured on Ny-t and Ny-p on day 3, when compared to control. Thus, the nature of the surface can directly influence MSCs differentiation, ultimately affecting the quality of new tissue formation with BOPP-t suppressing osteogenic differentiation. [Copyright &y& Elsevier]