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Chemical–physical and in vivo evaluations of a self-assembling amphiphilic peptide as an injectable hydrogel scaffold for biomedical applications

The self-aggregation and gelation of an amphiphilic peptide (C17H35CONH–A4G3ERGD, peptide amphiphile) were studied by light scattering, viscometry, nuclear magnetic resonance diffusometry, and atomic force microscopy. The peptide amphiphile critical aggregation concentration was evaluated to be 16 and 60 µM by light scattering and viscometry, respectively. The observed difference was attributed to the larger sensitivity of the latter technique to the presence of long fibrils. The addition of one equivalent or more of divalent cations (Ca2+ and Mg2+) to peptide amphiphile formed dense incoherent hydrogels. Based on the atomic force microscopy and nanoindentation data, both the hydrogel morphology and stiffness were independent of the cation type and peptide amphiphile concentration. However, gel stiffness increased on increasing Ca2+/peptide amphiphile molar ratio while a parallel decrease in the apparent water diffusion rate was observed by nuclear magnetic resonance diffusometry. The dispersions of endothelial progenitor cells in the peptide amphiphile hydrogels were evaluated in vivo on a rat tissue hypoxia model. Significant capillary formation at the injection site was observed by tissue appearance and histological examination, which indicated endothelial progenitor cell/peptide amphiphile hydrogel-enhanced angiogenesis in ischemic tissue.