An Engineered Biomimetic Peptide Regulates Cell Behavior by Synergistic Integrin and Growth Factor Signaling

Abstract

Recreating the healing microenvironment is essential to regulate cell–material interactions and ensure the integration of biomaterials. To repair bone, such bioactivity can be achieved by mimicking its extracellular matrix (ECM) and by stimulating integrin and growth factor (GF) signaling. However, current approaches relying on the use of GFs, such as bone morphogenetic protein 2 (BMP‐2), entail clinical risks. Here, a biomimetic peptide integrating the RGD cell adhesive sequence and the osteogenic DWIVA motif derived from the wrist epitope of BMP‐2 is presented. The approach offers the advantage of having a spatial control over the single binding of integrins and BMP receptors. Such multifunctional platform is designed to incorporate 3,4‐dihydroxyphenylalanine to bind metallic oxides with high affinity in a one step process. Functionalization of glass substrates with the engineered peptide is characterized by physicochemical methods, proving a successful surface modification. The biomimetic interfaces significantly improve the adhesion of C2C12 cells, inhibit myotube formation, and activate the BMP‐dependent signaling via p38. These effects are not observed on surfaces displaying only one bioactive motif, a mixture of both motifs or soluble DWIVA. These data prove the biological potential of recreating the ECM and engaging in integrin and GF crosstalk via molecular‐based mimics.

New trends in the development of multifunctional peptides to functionalize biomaterials

“…Recently, we have engineered a biomimetic multifunctional peptide containing the RGD and the DWIVA motifs in a spatially defined geometry and two DOPA molecules to anchor the molecule to model glass substrates (Figure 5C). 58 The biomimetic interface significantly increased C2C12 adhesion and spreading, and inhibited myotube formation, a wellknown indicator of BMP-2 activity. 128 Moreover, activation of BMPdependent signaling via p38 was observed.…”

“…72 (C) Branched peptide combining the RGD and DWIVA sequences. 58 Active sequences represented in red and green colors, spacers in yellow and anchorage units in blue peptide showed positive effects in reducing S. mutans biofilm formation and promoting MSC adhesion and mineralization. 106 However, the peptide was coated on the surfaces by simple physisorption and the biological effects were highly dependent on the chemistry of the surface used.…”

“…Most interestingly, integrins and BMP receptors have shown synergistic effects, opening the way to engineer novel surface functionalization techniques to enhance bioactivity of biomaterials. [56][57][58] • Preventing bacterial adhesion without loosening cell adhesive properties: in addition to mimicking ECM characteristics, bacterial colonization is another challenge. This is especially important in orthopedics and dental applications, in which metals, which are prone to bacterial infection, are commonly used.…”

Biocompatibility pathways and mechanisms for bioactive materials: The bioactivity zone

“…This has not been a straightforward process and few clear strategies have emerged with respect to cell-adhesive bioactive materials. Recent work on enhanced cellular behavior mediated by E-cadherin surface immobilization [ 70 ] and the regulation of cell behavior by synergistic integrin and growth factor signaling [ 71 ] show some promise, but the detailed observations are obtained during in vitro studies. Similarly, Barros et al were able to demonstrate the potential of mono-PEGylated recombinant human N-terminal agrin as a natural affinity binding ligand for the site-selective immobilization of laminin and resulting mediation of cell adhesion and spreading [ 72 ].…”

A versatile click chemistry-based approach for functionalizing biomaterials of diverse nature with bioactive peptides

“…The use of copper(I)-catalysed azide-alkyne cycloaddition reaction (CuAAc) on solid-phase allows the preparation of peptide mimetics containing specific anchoring units in a selective and stereospecific manner. 19,20 As a proof of concept, we selected a biomimetic multifunctional peptide, recently developed by us, 21 and focused on the incorporation of three different anchoring groups (thiol, catechol and amino) in solid-phase. To this end, suitable functional groups were incorporated in both moieties, i.e.…”