Production and Characterization of Recombinant Collagen-Binding Resilin Nanocomposite for Regenerative Medicine Applications

Abstract: Development of mechanically stable and multifunctional biomaterials for sensing, repair, and regeneration applications is of great importance. Herein, we investigate the potential of recombinant resilin-like (Res) nanocomposite elastomer as a template biomaterial for regenerative devices such as adhesive bandages or films, electrospun fibers, screws, sutures, and drug delivery vehicles. Exon I (Rec1) from the native resilin gene of Drosophila (CG15920) was fused with collagen-binding domain (ColBD) from Clostridium histolyticum and expressed in Komagataella pastoris (formerly Pichia pastoris). The 100% binding of Resilin-ColBD (Res-ColBD) to collagen I was shown at a 1:1 ratio by mass. Atomic force microscopy results in force mode show a bimodal profile for the ColBD-binding interactions. Moreover, based on the force-volume map, Res-ColBD adhesion to collagen was statistically significantly higher than resilin without ColBD. Lay Summary: Designing advanced biomaterials that will not only withstand the repetitive mechanical loading and flexibility of tissues but also retain biochemical and biophysical interactions remains challenging. The combination of physical, biological, and chemical cues is vital for disease regulation, healing, and ultimately complete regeneration of functional human tissues. Resilin is a super elastic and highly resilient natural protein with good biocompatibility but lacks specific biological and chemical cues. Therefore, resilin decorated with collagen I–binding domain is proposed as a functional nanocomposite template biomaterial. Collagen I is an ideal binding target, as it is the most abundant structural protein found in human body including scars that affect unwanted adhesion. Future Work: Musculoskeletal-related injuries and disorders are the second largest cause of disabilities worldwide. Significant pain, neurological discomfort, limited mobility, and substantial financial burden are associated with these disorders. Thus, biocompatible materials comprised of resilin with collagen-binding domain, such as films adhesive bandages (films, fiber matts, or hydrogels), sutures, screws and rods, three-dimensional scaffolds, and delivery vehicles, will be designed and evaluated for multiple musculoskeletal-related regeneration applications.