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.

Emergence of Elastic Properties in a Minimalist Resilin‐Derived Heptapeptide upon Bromination

“…Both Res-Br and Res-Wt were extremely soluble in water as expected since the parental Exon I sequence of Res is highly hydrophilic. [25] Circular dichroism (CD) spectra of 10 mm aqueous solutions of the two peptides revealed the signature of a polyproline II (PPII) conformation, [33] which is typical of peptide elastomers and consists of an intense negative band in the 190-200 nm range (Figure S13, Supporting Information). A small red shift of the negative peak in the CD spectrum with respect to that of Res-Wt suggested a possible role of bromine in altering the π→π* charge-transfer transition [34] related to the peptide bond.…”

“…Both Res‐Br and Res‐Wt were extremely soluble in water as expected since the parental Exon I sequence of Res is highly hydrophilic. [ 25 ]…”

“…In this frame, many efforts have been spent on the biosynthesis of Res-like polypeptides (RLPs), endowed with both native Res regions and heterologous domains, to shape new properties for potential biomedical applications, [21][22][23] including tissue engineering [24] and drug delivery, [25] as well as wearable sensors. [26][27][28][29] RLPs' elasticity arises from covalent cross-linking among polymer chains, which limits their extension and sliding, ensuring a complete shape recovery after deformation.…”

Identification and characterization of epicuticular proteins of nematodes sharing motifs with cuticular proteins of arthropods

“…Innovative ways to produce epicuticlins are needed. Recombinant epicuticlins would be especially useful for future applications in the biomaterial sciences, as shown for recombinant resilin [ 75 ] and elastin [ 76 ]. The potential of the highly organized characteristics of epicuticlins is evident, for example, as biosynthetic polymers in medical applications, such as in skin healing procedures.…”

Global Biomaterials Market: Potential Opportunities for Raw Materials of Animal Origin

“…Для улучшения свойств имплантируемых материалов на основе коллагена и придания им большей механической прочности предложено получать композиты коллагена с керамиками и синтетическими полимерами (полиэтиленом, поливиниловым спиртом, полисилоксанами). Композиты коллагена и гидроксиапатита рассматриваются в качестве остеозамещающего материала для восстановления дефектов костной ткани в челюстно-лицевой хирургии и стоматологии [37][38][39][40][41][42][43][44].…”