Three-dimensional (3D) porous scaffolds combined with therapeutic stem cells play vital roles in tissue engineering. The adult brain has very limited regeneration ability after injuries such as trauma and stroke. In this study, injectable 3D silk fibroin-based hydrogel scaffolds with encapsulated neural stem cells were developed, aiming at supporting brain regeneration. To improve the function of the hydrogel towards neural stem cells, silk fibroin was modified by an IKVAV peptide through covalent binding. Both unmodified and modified silk fibroin hydrogels were obtained, through sonication, with mechanical stiffness comparable to that of brain tissue. Human neural stem cells were encapsulated in both hydrogels and the effects of IKVAV peptide conjugation on cell viability and neural differentiation were assessed. The silk fibroin hydrogel modified by IKVAV peptide showed increased cell viability and an enhanced neuronal differentiation capability, which contributed to understanding the effects of IKVAV peptide on the behaviour of neural stem cells. For these reasons, IKVAV-modified silk fibroin is a promising material for brain tissue engineering. Copyright (C) 2015 John Wiley & Sons, Ltd.

Sufficient vascularization of the implant construct is required for tissue regeneration to ensure the supply of oxygen and nutrients. In our previous work, we established sonication-induced silk fibroin hydrogel to load neural stem cells for brain tissue engineering applications. In this study, we explored the application of silk fibroin as an injectable hydrogel for vascularization of soft tissues. We investigated the ability of outgrowth endothelial cells (OECs) in mono-culture or in co-culture with human bone marrow-derived mesenchymal stem cells (BM-MSCs) to form capillary networks in silk fibroin hydrogels. Furthermore, the silk fibroin hydrogel was modified with IKVAV peptide revealing a sequence derived from the extracellular matrix component laminin-1 to test its effects on angiogenesis, using unmodified and VVIAK modified silk fibroin hydrogel as controls. In monocultures of OECs, no angiogenic structures were observed in silk fibroin hydrogels. In contrast, vascular structures were abundant and increased in co-culture, as confirmed by immunocytochemistry and scanning electron microscopy (SEM) over 10 d of culture in silk fibroin-based hydrogels. Although no significant differences in angiogenic activity seem to be caused by the IKVAV peptide in our experimental settings, these results indicate that sonication-induced silk fibroin-based hydrogels support the formation of functional endothelial tubes and vascularization networks in the presence of mesenchymal cells supporting the vascular sprouting of endothelial cells.

Different hydrogel materials have been prepared to investigate the effects of culture substrate on the behaviour of pluripotent cells. In particular, genipin‐crosslinked gelatin–silk fibroin hydrogels of different compositions have been prepared, physically characterized and used as substrates for the culture of pluripotent cells. Pluripotent cells cultured on hydrogels remained viable and proliferated. Gelatin and silk fibroin promoted the proliferation of cells in the short and long term, respectively. Moreover, cells cultured on genipin‐crosslinked gelatin–silk fibroin blended hydrogels were induced to an epithelial ectodermal differentiation fate, instead of the neural ectodermal fate obtained by culturing on tissue culture plates. This work confirms that specific culture substrates can be used to modulate the behaviour of pluripotent cells and that our genipin‐crosslinked gelatin–silk fibroin blended hydrogels can induce pluripotent cells differentiation to an epithelial ectodermal fate. Copyright © 2014 John Wiley & Sons, Ltd.