Motivated by the outstanding mechanical properties of natural resilin, a recombinant resilin-like polypeptide (RLP12) has been expressed, purified and cross-linked into hydrogels via a Michael-type addition reaction.  The recursive ligation of our original RLP12 gene has led to three new RLPs withincreased cysteine functionality. The cloning, expression and purification of these proteins were verified via gene sequencing, SDS-PAGE, MALDI mass spectroscopy and amino acid analysis. These RLP hydrogels can be cross-linked by reaction of cysteine residues with a multi-arm star PEG terminated with vinyl sulfone; this strategy has led to homogenous, stable networks and has provided the opportunity for 3D cell encapsulation. Oscillatory rheology was employed to study the mechanical properties of the RLP-PEG hydrogels and LIVE/DEAD demonstrated the viability of encapsulated NIH3T3 murine fibroblasts and human aortic adventitial fibroblasts. These resilin-based protein-polymer hydrogels offer a promising strategy to the development of cell-instructive tissue engineering scaffolds for mechanically active tissues.

Resilin-PEG hybrid hydrogels as tissue engineering scaffolds for cardiovascular tissues

244th National Meeting (2012)

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