A new bottom-up strategy based on aromatic peptide amphiphile is developed for a high-contrast visualization of 3D live cell-material imaging-something that has been difficult to achieve previously because of the problems associated with the diffraction of light by the nanosized peptide materials and the aggregation-caused quenching of aggregated π-conjugated fluorophores in the nanostructures. This study reports an example of a novel supramolecular hydrogelator, naphthaleneimide-phenylalanine (NI-Phe), which forms a self-supporting hydrogel displaying a unique microfibrous network and promising aggregation-induced emission characteristics at pH 7.4. The storage modulus of the NI-Phe gel supports the mass of a cell for 3D cell culturing. This work illustrates a new dopant-free supramolecular approach, complementary to well-established doping procedures that should facilitate the development of live cell imaging in 3D scaffolding materials.

Fluorenyl-9-methoxycarbonyl (Fmoc)-diphenylalanine (Fmoc-FF) and Fmoc-arginine-glycine--aspartate (Fmoc-RGD) peptides self-assemble to form a 3D network of supramolecular hydrogel (Fmoc-FF/Fmoc-RGD), which provides a nanofibrous network that uniquely presents bioactive ligands at the fiber surface for cell attachment. In the present study, mesenchymal stem cells (MSCs) in Fmoc-FF/Fmoc-RGD hydrogel increase in proliferation and survival compared to those in Fmoc-FF/Fmoc-RGE hydrogel. Moreover, MSCs encapsulated in Fmoc-FF/Fmoc-RGD hydrogel and induced in each defined induction medium undergo in vitro osteogenic, adipogenic, and chondrogenic differentiation. For in vivo differentiation, MSCs encapsulated in hydrogel are induced in each defined medium for one week, followed by injection into gelatin sponges and transplantation into immunodeficient mice for four weeks. MSCs in Fmoc-FF/Fmoc-RGD hydrogel increase in differentiation into osteogenic, adipogenic, and chondrogenic differentiation, compared to those in Fmoc-FF/Fmoc-RGE hydrogel. This study concludes that nanofibers formed by the self-assembly of Fmoc-FF and Fmoc-RGD are suitable for the attachment, proliferation, and multi-differentiation of MSCs, and can be applied in musculoskeletal tissue engineering.

Due to the replacement of only one hydrogen atom by fluorine, 4-fluorobenzyl-diphenylalanine molecules self-assemble to form a transparent hydrogel.