Replicating the multi-hierarchical self-assembly of collagen has long-attracted scientists, from both the perspective of the fundamental science of supramolecular chemistry and that of potential biomedical applications in tissue engineering. Many approaches to drive the self-assembly of synthetic systems through the same steps as those of natural collagen (peptide chain to triple helix to nanofibres and, finally, to a hydrogel) are partially successful, but none simultaneously demonstrate all the levels of structural assembly. Here we describe a peptide that replicates the self-assembly of collagen through each of these steps. The peptide features collagen's characteristic proline-hydroxyproline-glycine repeating unit, complemented by designed salt-bridged hydrogen bonds between lysine and aspartate to stabilize the triple helix in a sticky-ended assembly. This assembly is propagated into nanofibres with characteristic triple helical packing and lengths with a lower bound of several hundred nanometres. These nanofibres form a hydrogel that is degraded by collagenase at a similar rate to that of natural collagen.

A series of nine, frustrated, multidomain peptides is described in which forces favoring self-assembly into a nanofiber versus those favoring disassembly could be easily modified. The peptides are organized into an ABA block motif in which the central B block is composed of alternating hydrophilic and hydrophobic amino acids (glutamine and leucine, respectively). This alternation allows the amino acid side chains to segregate on opposite sides of the peptide backbone when it is in a fully extended beta-sheet conformation. In water, packing between two such peptides stabilizes the extended conformation by satisfying the desire of the leucine side chains to exclude themselves from the aqueous environment. Once in this conformation intermolecular backbone hydrogen bonding can readily take place between additional peptides eventually growing into high aspect ratio fibers. B block assembly may continue infinitely or until monomeric peptides are depleted from solution which results in an insoluble precipitate. Block A consists of a variable number of positively charged lysine residues whose electrostatic repulsion at pH 7 works against the desire of the B block to assemble. Here we show that balancing the forces of block A against B allows the formation of controlled length, individually dispersed, and fully soluble nanofibers with a width of 6 +/- 1 nm and length of 120 +/- 30 nm. Analysis by infrared, circular dichroism, and vitreous ice cryo-transmission electron microscopy reveals that the relative sizes of blocks A and B dictate the peptide secondary structure which in turn controls the resulting nanostructure. The system described epitomizes the use of molecular frustration in the design of finite self-assembled structures. These materials, and ones based on their architecture, may find applications where nanostructured control over fiber architecture and chemical functionality is required.

Peptide hydrogels show immense promise as therapeutic materials. Here we present a rationally designed multidomain peptide that self-assembles into nanofibers approximately 8 nm wide, 2 nanometers high and microns in length in the presence of Mg2+. At a concentration of 1% by weight, the peptide forms an extensive nanofibers network that results in a physically cross-linked viscoelastic hydrogel. This hydrogel undergoes shear thinning and then quickly recovers nearly 100% of its elastic modulus when the shearing force is released, making it ideal for use as an injectable material. When placed in the presence of human embryonic stem cells (ESCs), the nanofibrous hydrogel acts like a sponge, soaking up the vast array of growth factors and cytokines released by the ESCs. The peptide hydrogel sponge can then be removed from the presence of the ESCs and placed in a therapeutic environment, where it can subsequently release these components. In vitro experiments demonstrate that release of stem cell secretome from these hydrogels in the presence of Glomerular Epithelial Cells treated with high glucose significantly decreased protein permeability in a model of diabetes-induced kidney injury. Tracking experiments were then performed to determine the fate of the hydrogel upon injection in vivo. Hydrogels labeled with a Gd3+ MRI contrast agent were injected into the abdominal cavity of mice and found to remain localized over twenty-four hours. This implies that the hydrogel possesses sufficient rigidity to remain localized and release stem cell secretome over time rather than immediately dissolving in the abdominal cavity. Together, the shear thinning and recovery as observed by rheometry, as well as secretome absorption and release in vivo, demonstrate the potential of the nanofibrous multidomain peptide hydrogel as an injectable delivery agent.

Self-assembling multidomain peptides have been shown to have desirable properties, such as the ability to form hydrogels that rapidly recover following shear-thinning and the potential to be tailored by amino acid selection to vary their elasticity and encapsulate and deliver proteins and cells. Here we describe the effects of substitution of aliphatic hydrophobic amino acids in the central domain of the peptide for the aromatic amino acids phenylalanine, tyrosine and tryptophan. While the basic nanofibrous morphology is retained in all cases, selection of the particular core residues results in switching from anti-parallel hydrogen bonding to parallel hydrogen bonding in addition to changes in nanofiber morphology and in hydrogel rheological properties. Peptide nanofiber assemblies are investigated by circular dichroism polarimetry, infrared spectroscopy, atomic force microscopy, transmission and scanning electron microscopy, oscillatory rheology and molecular dynamics simulations. Results from this study will aid in designing next generation cell scaffolding materials.

Organogels obtained from plant wax and soybean oil were tested for their suitability for incorporation into margarine. Sunflower wax, rice bran wax and candelilla wax were evaluated. Candelilla wax showed phase separation after making the emulsion with the formulation used in this study. Rice bran wax showed relatively good firmness with the organogel, but dramatically lowered firmness for a margarine sample. Sunflower wax showed the greatest firmness for organogel and the margarine samples among the three plant waxes tested in this study. Firmness of the margarine containing 2-6 % sunflower wax in soybean oil was similar to that of margarine containing 18-30 % hydrogenated soybean oil in soybean oil. The firmness of commercial spread could be achieved with about 2 % sunflower wax and that of commercial margarine could be achieved with about 10 % of sunflower wax in the margarine formulation. Dropping point, DSC and solid fat content of the new margarine containing 2-6 % sunflower wax showed a higher melting point than commercial margarine and spreads.

A series of self-assembling multidomain peptides have been designed, synthesized, and tested for their ability to individually suspend single-walled carbon nanotubes (SWCNTs) in water while preserving strong near-IR nanotube luminescence. Photometric and spectral measurements on individual SWCNTs revealed that emission in the common biocompatible coating agents Pluronic F127, ss-DNA, and BSA is approximately an order of magnitude weaker than in the bio-incompatible ionic surfactant SDBS. By contrast, one of the engineered peptides gave SWCNT emission ~40% as intense as in SDBS. A strong inverse correlation was also found between the spectral line widths of coated SWCNTs and the efficiency of their emission. Peptides with rationally designed selfassembly properties appear to be promising coatings that may enable SWCNT optical sensing applications in biological environments.

This study showed that sunflower wax could be used as an alternative to traditional solid fats for the development of new margarine and spread products from a variety of healthy vegetable oils.

The rheological properties of the environment in which a cell lives play a key role in how the cells will respond to that environment and may modify cell proliferation, morphology and differentiation. Effective means of modifying these properties are needed, particularly for peptide hydrogels which are generally relatively weak and soft. In this report we describe the enzymatic cross-linking of a nanofibrous multidomain peptide hydrogel. When this method was used, the storage modulus, G', could be increased to over 4000 Pa without changes in hydrogel concentration and without dramatic changes in nanostructural architecture. Enzymatic cross-linking represents a mild and simple method for increasing the mechanical strength of peptide hydrogels in applications for which the robustness of the gel is essential. This method should be suitable for a broad array of peptide hydrogels containing lysine such as those currently under study by many different groups.