Paciello, Antonio (2014) Peptide-conjugated hydrogels for cell and tissue recognition. [Tesi di dottorato]

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The development of biomaterials has recently focused on the design and synthesis of peptide-conjugated hydrogels that are capable of eliciting some specific responses to cell and tissue mediated by biomolecular recognition, which can be manipulated by altering design of the material. Biomolecular recognition of hydrogel materials by cells and tissue has been achieved by surface and bulk modification via chemical or physical methods, with bioactive molecules as short peptide sequences that can incur specific interactions with cell receptors or tissue binding-site. Peptides, indeed, are the smallest bioactive entities contained in a protein and they are responsible, in particular way, of full chemical interactions and spatial arrangement, which together supply the biological recognitions. Hydrogels, instead, represent one the most popular materials used in tissue engineering and regenerative medicine for their versatile properties. Here, are reported the preparation of DVS-crosslinked hyaluronic acid-based hydrogels for tissue wound dressing applications and the synthesis of novel supramolecular PEI-based hydrogels as cell microenvironments for biomedical applications. In the first part, collagen type I binding fragment conjugated to microbeads of hyaluronic acid have shown non-covalent adhesive interactions on human derma and cornea of eye, after mechanical or photo-removal of epithelial tissues. Were evaluated microbeads adhesive properties on human derma and cornea of eye, and was observed that they formed a continuous covering layer from indiscriminately biological provenience of the collagen type I. This novel instructed biomaterial show abilities to cover damaged tissue only where was provoked the injury, as well as the final layer was originated by deformation of many micro-entities capable to arrange itself until to completely dress the injury regions. In second part, novel PEI hydrogels were synthesized, they showed easily chemical activation by multi-photon laser irradiation showing interesting three-dimensional patterning capabilities in presence of peptide as RGD and biomolecules with free carboxylic acid or hydroxyl groups without use of adjunctive photo-initiators or activators. PEI hydrogels activated for photo-conjugation can be indispensable to the spatial distribution control of bioactive signal, which represents an important factor for cell recognition and cell-guidance behaviour in tissue genesis and regeneration. This project is today the first study on the supramolecular hydrogels activated for photo-conjugation chemistry, and can represent a novel cell-instructive platform that can be selectively encoded with active signals, with relevant applications in biotechnology and medicine.

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