No abstract

This work presents an environmentally friendly, iodine-catalysed chemical modification method to generate highly hydrophobic, optically active cellulose nanocrystals (CNC). The high degree of ester substitution (DS=2.18), hydrophobicity, crystalline behaviour and optical activity of the generated acetylated CNC (Ac-CNC) were quantified by TEM, FTIR, solid 13C NMR, contact angle, XRD and POM analyses. Ac-CNC possessing substantial enhancement in thermal stability (16.8%) and forms thin films with interlayer distance of 50-150 nm, presenting cavities suitable for entrapping nano and micro particles. Generated Ac-CNC proved as an effective reinforcing agent in hydrophobic polymer matrices for fabricating high performance nanocomposites. When integrated at a very low weight percentage (0.5%) in an epoxy matrix, Ac-CNC provided for a 73% increase in tensile strength and a 98% increase in modulus, demonstrating its remarkable reinforcing potential and effective stress transfer behaviour. The method of modification and the unique properties of the modified CNC (hydrophobicity, crystallinity, reinforcing ability and optical activity) render them a novel bionanomaterial for a range of multipurpose applications.

Resilin is an elastomeric protein found in specialized regions of the cuticle of most insects, providing outstanding material properties including high resilience and fatigue lifetime for insect flight and jumping needs. Two exons (1 and 3) from the resilin gene in Drosophila melanogaster were cloned and the encoded proteins expressed as soluble products in Escherichia coli. A heat and salt precipitation method was used for efficient purification of the recombinant proteins. The proteins were solution cast from water and formed into rubber-like biomaterials via horseradish peroxidase-mediated cross-linking. Comparative studies of the two proteins expressed from the two different exons were investigated by Fourier Transform Infrared Spectroscopy (FTIR) and Circular Dichrosim (CD) for structural features. Little structural organization was found, suggesting structural order was not induced by the enzyme-mediateed dityrosine cross-links. Atomic Force Microscopy (AFM) was used to study the elastomeric properties of the uncross-linked and cross-linked proteins. The protein from exon 1 exhibited 90% resilience in comparison to 63% for the protein from exon 3, and therefore may be the more critical domain for functional materials to mimic native resilin. Further, the cross-linking of the recombinant exon 1 via the citrate-modified photo-Fenton reaction was explored as an alternative dityrosine mediated polymerization method and resulted in both highly elastic and adhesive materials. The citrate-modified photo-Fenton system may be suitable for in-vivo applications of resilin biomaterials.

This research explores the properties of bionanocomposite films prepared by binding recombinant resilin-like protein (res) consisting of the exon 1 resilin sequence from Drosophila melanogaster engineered to include a cellulose binding domain (CBD), to cellulose nanocrystals (CNCs). The optimal binding of res-CBD to CNCs was 1:5 by mass, and the resulting res-CBD-CNCs remained colloidally stable in water. Res-CBD-CNCs were solvent cast into transparent, free-standing films, which were more hydrophobic than neat CNC films, with water contact angles of 70-80°compared to 35-40°for the latter. In contrast to the multi-domain orientation typical of chiral nematic CNC films, res-CBD-CNC and CBD-CNC films exhibited long-range, uniaxial orientation that was apparently driven by the CBD moiety. Glycerol was studied as an additive in the films to determine whether the addition of a wet component to solvate the recombinant protein improved the mechanical properties of the res-CBD-CNC films. In comparison to the other films, res-CBD-CNC films were more elastic with added glycerol, demonstrating a range of 0.5-5 wt% (i.e., the films responded more elastically to a given strain and/or were less plastically deformed by a given mechanical load), but became less elastic with added glycerol between 0.5-5 wt%. Overall, films made of res-CBD-CNCs plus 0.5 wt% glycerol displayed improved mechanical properties compared to neat CNC films, and with an increase in toughness of 150% and in elasticity of 100%.

Collagen type I, in various physical forms, is widely used in tissue engineering and regenerative medicine. To control the mechanical properties and biodegradability of collagen-based devices, exogenous cross-links are introduced into the 3D supramolecular structure. However, potent cross-linking methods are associated with cytotoxicity, whilst mild cross-linking methods are associated with suboptimal mechanical resilience. Herein, we assessed the influence of resilin, a super-elastic and highly stretchable protein found within structures in arthropods where energy storage and long-range elasticity are needed, on the biophysical and biological properties of mildly cross-linked extruded collagen fibres. The addition of resilin-like protein in the 4-arm poly(ethylene glycol) ether tetrasuccinimidyl glutarate cross-linked collagen fibres resulted in a significant increase of stress and strain at break values and a significant decrease of modulus values. The addition of resilin-like protein did not compromise cell metabolic activity and DNA concentration. All groups are supported parallel to the longitudinal fibre axis cell orientation. Herein we provide evidence that the addition of resilin-like protein in mildly cross-linked collagen fibres improves their biomechanical properties, without jeopardising their biological properties.