(502c) Characterization of Modular Resilin-Based Artificial Protein Matrices for Cartilage Tissue Engineering

Recombinant proteins have been explored for numerous tissue engineering applications. In particular, artificial proteins based on structural repeats of elastin and silk have been investigated for use in soft tissue and orthopedic applications. Compared to synthetic or natural materials, recombinant proteins have a number of advantages that include: exquisite control over sequence and composition; precise molecular weight; and a modular nature, which allows for the functional domains to be easily altered.

We are developing an artificial protein matrix based on a structural repeat found in resilin, an elastic protein from insect joints and tendons that contributes to flight and jumping. Resilin has a high strain and can efficiently store energy. In addition, it efficiently recovers after deformation and has a high fatigue lifetime, which makes it ideal for use in repetitive environments. Based on the natural properties of resilin, we believe that these resilin-based proteins are promising candidates for cartilage engineering. Our modular protein design includes bioactive domains and sites for either chemical or photochemical crosslinking. We adapted a recursive cloning technique and show that the number of resilin repeats in our protein can be precisely and easily tuned. The Studier auto-induction method was used to express the proteins, but had to be modified due to protein degradation. Protein expression and purity are assessed by Western blot, amino acid analysis, and mass spectrometry.