Bioactive hydrogels based on Designer Collagens

Abstract

Designer Collagens are based on streptococcal collagen-like (Scl) proteins that form a triple helix similar to mammalian collagens but that are non-platelet aggregating. In contrast to the numerous cell-binding sites on collagen, Scl2 from Streptococcus pyogenes serotype M28 does not contain any known cell-binding sites and thus provides a blank slate in terms of cellular interactions. In the current study, Scl2 protein was modified to include receptor binding motifs that interact with alpha1 and/or alpha2 integrin subunits. The modfied Scl2 proteins have been demonstrated to mediate differential endothelial cell (EC) and smooth muscle cell (SMC) adhesion via these integrins and to retain the non-platelet aggregating properties of the "parent" Scl2. Thromboresistant scaffolds which selectively bind ECs vs. SMCs would be desirable for vascular repair or replacement. Despite the potential of these Scl proteins in vascular applications, the utility of this recombinant protein family is currently limited to coatings due to the inability of Scl proteins to assemble into stable three-dimensional networks. To address this limitation, the Scl2 proteins were functionalized with photocrosslinking sites to enable incorporation into a hydrogel matrix. Characterization studies confirmed that the functionalization of the Scl2 proteins did not disrupt triple helix conformation, integrin binding or cell adhesion. Bioactive hydrogels were fabricated by combining the functionalized Scl2 proteins with poly(ethylene glycol) diacrylate (PEGDA) and photocrosslinking. EC and SMC adhesion studies confirmed cell-specific adhesion due to selective integrin binding to the two receptor binding motifs investigated. These results serve to highlight the potential of this novel biomaterial platform in the development of improved tissue engineered vascular grafts.