Enhanced cell-material interactions through the biofunctionalization of polymeric surfaces with engineered peptides.

Research on surface modification of polymeric materials to guide the cellular activity in biomaterials designed for tissue engineering applications has mostly focused on the use of natural extracellular matrix (ECM) proteins and short peptides, such as RGD. However, the use of engineered proteins can gather the advantages of these strategies and avoid the main drawbacks. In this study, recombinant engineered proteins called elastin-like recombinamers (ELRs) have been used to functionalize poly(lactic) acid (PLA) model surfaces. The structure of the ELRs has been designed to include the integrin ligand RGDS and the cross-linking module VPGKG. Surface functionalization has been characterized and optimized by means of ELISA and atomic force microscopy (AFM). The results suggest that ELR functionalization creates a nonfouling canvas able to restrict unspecific adsorption of proteins. Moreover, AFM analysis reveals the conformation and disposition of ELRs on the surface. Biological performance of PLA surfaces functionalized with ELRs has been studied and compared with the use of short peptides. Cell response has been assessed for different functionalization conditions in the presence and absence of the bovine serum albumin (BSA) protein, which could interfere with the surface-cell interaction by adsorbing on the interface. Studies have shown that ELRs are able to elicit higher rates of cell attachment, stronger cell anchorages and faster levels of proliferation than peptides. This work has demonstrated that the use of engineered proteins is a more efficient strategy to guide the cellular activity than the use of short peptides, because they not only allow for better cell attachment and proliferation, but also can provide more complex properties such as the creation of nonfouling surfaces.

Biofunctionalization of polymeric surfaces.

Most of the synthetic polymeric biomaterials used for biomedical applications lack of functional groups able to specifically instruct cells to unlock their potential for tissue regeneration. Surface modification strategies are able to overcome this limitation by introducing bioactive cues. In this study, several functionalization approaches are analyzed. Wet chemical methods such as controlled hydrolysis of polyesters followed by biomolecules grafting by carbodiimide chemistry are simple and versatile approaches, able to succesfully improve the bioactivity of devices with virtually any architecture. Grafting of short peptides, extracellular matrix proteins (ECM) or engineered protein-like recombinamers are promising techniques to improve cell adhesion to biomaterials, including polylactic acid (PLA) and its derivatives. ECM molecules and recombinamers can present more effectively bioactive signals, even in presence of competing, nonadhesive serum proteins. Besides adhesion, surface modifications intended to improve cell attachment, play a role on other cell responses, such as migratory potential. Collagen coating were shown to enhance the expression of the migratory receptor CXCR4 in mesenchymal stromal cells, when compared to short RGD peptides, while the modality of functionalization (covalent vs. physisorbed) tuned the rate of cell migration from PLA-based microcarriers. This multiple effects have to be taken into account when designing biomaterials for cell delivery and tissue engineering. Furthermore, as we aim to recapitulate in vitro the complexity of native tissues, alternative strategies based on the generation of decellularized polymer scaffold rich in cell-deposited ECM are proposed.

Biomolecular functionalization for enhanced cell-material interactions of poly(methyl methacrylate) surfaces.

The integration of implants or medical devices into the body tissues requires of good cell-material interactions. However, most polymeric materials used for these applications lack on biological cues, which enhanced mid- and long-term implant failure due to weak integration with the surrounding tissue. Commonly used strategies for tissue-material integration focus on functionalization of the material surface by means of natural proteins or short peptides. However, the use of these biomolecules involves major drawbacks such as immunogenic problems and oversimplification of the constructs. Here, designed elastin-like recombinamers (ELRs) are used to enhance poly(methyl methacrylate) surface properties and compared against the use of short peptides. In this study, cell response has been analysed for different functionalization conditions in the presence and absence of a competing protein, which interferes on surface-cell interaction by unspecific adsorption on the interface. The study has shown that ELRs can induce higher rates of cell attachment and stronger cell anchorages than short peptides, being a better choice for surface functionalization.