Diversity of viscoelastic properties of an engineered muscle-inspired protein hydrogel

Folded protein hydrogels are prime candidates as tuneable biomaterials but it is unclear to what extent their mechanical properties have mesoscopic, as opposed to molecular origins. To address this, we probe hydrogels inspired by the muscle protein titin and engineered to the polyprotein I275, using a multimodal rheology approach. Across multiple protocols, the hydrogels consistently exhibit power-law viscoelasticity in the linear viscoelastic regime with an exponent β = 0.03, suggesting a dense fractal meso-structure, with predicted fractal dimension df = 2.48. In the nonlinear viscoelastic regime, the hydrogel undergoes stiffening and energy dissipation, indicating simultaneous alignment and unfolding of the folded proteins on the nanoscale. Remarkably, this behaviour is highly reversible, as the value of β, df and the viscoelastic moduli return to their equilibrium value, even after multiple cycles of deformation. This highlights a previously unrevealed diversity of viscoelastic properties that originate on both at the nanoscale and the mesoscopic scale, providing powerful opportunities for engineering novel biomaterials

Gels formed from amino-acid derivatives, their novel rheology as probed by bulk and particle tracking rheological methods

This is the final version of the article. It first appeared from Elsevier via http://dx.doi.org/10.1016/j.jnnfm.2014.09.008We discuss the use of dynamic light scattering based particle micro-rheology to probe the lengthscale dependence of the microstructures formed by Fmoc-tyrosine gels. Past studies on these systems using dye diffusion have shown that Fmoc-tyrosine is capable of forming gels that can entrap molecules if they are large enough, unlike those gels formed by Fmoc-phenylalanine (Sutton et al., 2009). This result seems at odds with microscopic studies of the gel microstructure, which indicate porosity on much larger lengthscales than the molecular probes used. Here, we use particle probe based micro-rheology to investigate the porosity of the gels on larger lengthscales than is possible using molecular diffusion studies and show that there is considerable evidence of larger scale structures present in the gel. In particular we see that at no point does particle probe based micro-rheology reproduce the bulk properties of the gels, and also that there is strong dependence of the probe behaviour on particle size. Both of these results indicate the presence of microstructural features in the gel that are of the order of the particle size.Parts of this work are drawn from the doctoral thesis of A.A-R, for which the authors gratefully acknowledge funding from the EPSRC and Unilever R&D Colworth through an EPSRC CASE studentship

Reaction Rate Governs the Viscoelasticity and Nanostructure of Folded Protein Hydrogels

Hydrogels constructed from folded protein domains are of increasing interest as resilient and responsive biomaterials, but their optimization for applications requires time-consuming and costly molecular design. Here, we explore a complementary approach to control their properties by examining the influence of crosslinking rate on the structure and viscoelastic response of a model hydrogel constructed from photochemically crosslinked bovine serum albumin (BSA). Gelation is observed to follow a heterogeneous nucleation pathway in which BSA monomers crosslink into compact nuclei that grow into fractal percolated networks. Both the viscoelastic response probed by shear rheology and the nanostructure probed by small-angle X-ray scattering (SAXS) are shown to depend on the photochemical crosslinking reaction rate, with increased reaction rates corresponding to higher viscoelastic moduli, lower fractal dimension, and higher fractal cluster size. Reaction rate-dependent changes are shown to be consistent with a transition between diffusion- and rate-limited assembly, and the corresponding changes to viscoelastic response are proposed to arise from the presence of nonfractal depletion regions, as confirmed by SAXS. This controllable nanostructure and viscoelasticity constitute a potential route for the precise control of hydrogel properties, without the need for molecular modification

PE and PS Lipids Synergistically Enhance Membrane Poration by a Peptide with Anticancer Properties

Polybia-MP1 (MP1) is a bioactive host-defense peptide with known anticancer properties. Its activity is attributed to excess serine (phosphatidylserine (PS)) on the outer leaflet of cancer cells. Recently, higher quantities of phosphatidylethanolamine (PE) were also found at these cells' surface. We investigate the interaction of MP1 with model membranes in the presence and absence of POPS (PS) and DOPE (PE) to understand the role of lipid composition in MP1's anticancer characteristics. Indeed we find that PS lipids significantly enhance the bound concentration of peptide on the membrane by a factor of 7-8. However, through a combination of membrane permeability assays and imaging techniques we find that PE significantly increases the susceptibility of the membrane to disruption by these peptides and causes an order-of-magnitude increase in membrane permeability by facilitating the formation of larger transmembrane pores. Significantly, atomic-force microscopy imaging reveals differences in the pore formation mechanism with and without the presence of PE. Therefore, PS and PE lipids synergistically combine to enhance membrane poration by MP1, implying that the combined enrichment of both these lipids in the outer leaflet of cancer cells is highly significant for MP1's anticancer action. These mechanistic insights could aid development of novel chemotherapeutics that target pathological changes in the lipid composition of cancerous cells