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Development and in vitro biocompatibility assessment of novel supramolecular hydrogel biomaterial

Hydrogel biomaterials, closely mimicking the three-dimensional extracellular matrix, are considered to be an ideal material for cell and tissue scaffolding applications. The dynamic nature of the noncovalent hydrogels fibril network allows the hydrogel material to spontaneously adjust to the surrounding environment and the cells to migrate through the matrix comparing to covalent hydrogels whose pores are chemically constrained and relatively inflexible. A new type of such supramolecular gel has been designed and synthesized. The self-assembling tripeptide hydrogelator Ac-FFA-NH2 was prepared using classical methods of solution-state protein synthesis. Prepared peptide-type hydrogelator forms fibers by self-assembly giving stable hydrogel. Organization in gel assemblies at the supramolecular level was determined using spectroscopic methods and gel morphology by electronic microscopy. It is important to stress that the peptide three- dimensional gel network is held together by non-covalent interactions eg. hydrogen bonding, hydrophobic interactions and π-π stacking thus implying to build supramolecular fibril structures differently from conventional polymer materials. Series of experiments have been performed in order to assess in vitro biocompatibility of the novel material. HEK293T cells have been used as a reproducible model cell line and were cultured following established proliferation protocols. Survival and proliferation of HEK293T cells encapsulated within three- dimensional network of nanofibers formed by self-assembly of peptide amphiphile molecules have been evaluated. Results showed high viability and high proliferation rate of the cells distributed throughout the supramolecular hydrogel structure compared to Matrigel. The obtained material was proved to serve as stabile and biocompatible physical support in improving HEK293T cells biological outcome in vitro. Furthermore, these results suggest further evaluation of in vitro biocompatibility and bioactivity on the neural stem cells upon encapsulation in order to be investigated for possible brain tissue engineering application. Acknowledgments: The study was supported by EU FP7 grant GlowBrain (REGPOT–2012–CT2012–316120) and by the Croatian Science Foundation (HRZZ No. 7387).

self-assembling peptide; characterization; in vitro; biocompatibility

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