Design of a hydrophobic tripeptide that self-assembles into amphiphilic superstructures forming a hydrogel biomaterial.

We report the rational design of a heterochiral hydrophobic tripeptide self-assembling into amphiphilic d-superstructures that yield a self-supportive hydrogel at physiological pH. The material endures cell culture conditions and sustains fibroblast proliferation. Tripeptide superstructures are thoroughly analysed by several techniques.

Higher and lower supramolecular orders for the design of self-assembled heterochiral tripeptide hydrogel biomaterials.

The self-assembly behaviour of the eight stereoisomers of Val-Phe-Phe tripeptides under physiological conditions is assessed by several spectroscopy and microscopy techniques. We report the first examples of self-organised hydrogels from tripeptides in the l-d-l or d-l-d configuration, besides the expected gels with the d-l-l or l-d-d configuration, thus widening the scope for using amino acid chirality as a tool to drive self-assembly. Importantly, the positions of d- and l-amino acids in the gelling tripeptides determine a higher or lower supramolecular order, which translates into macroscopic gels with different rheological properties and thermal behaviours. The more durable hydrogels perform well in cytotoxicity assays, and also as peptides in solution. An appropriate design of the chirality of self-assembling sequences thus allows for the fine-tuning of the properties of the gel biomaterials. In conclusion, this study adds key details of supramolecular organization that will assist in the ex novo design of assembling chiral small molecules for their use as biomaterials.

Chirality effects at each amino acid position on tripeptide self-assembly into hydrogel biomaterials.

Hydrogels formed by ultrashort peptides are emerging as cost-effective materials for cell culture. However, L-peptides are labile to proteases, while their D-isomers are thought to not support cell growth as well. In contrast, the self-assembly behaviour and biological performance of heterochiral peptides (i.e., made of both d and l amino acids) are largely unknown. In this study, we evaluate the effects of amino acid chirality on tripeptide self-assembly and hydrogelation at physiological pH, and cytocompatibility in fibroblast cell culture. A series of uncapped hydrophobic tripeptides with all combinations of d, l amino acids was prepared, tested for self-assembly under physiological conditions, and analysed by circular dichroism, FT-IR, cryo-TEM, AFM, and Thioflavin T fluorescence imaging. Amino acid chirality has a profound effect on the peptides' supramolecular behaviour. Only selected isomers form hydrogels, and of amyloid structure, as confirmed by rheology and XRD. Importantly, they are able to maintain the viability and proliferation of fibroblasts in vitro. This study identifies two heterochiral gels that perform well in cell culture and will assist in the design of innovative and cost-effective peptide gel biomaterials.

In vitro fibroblast response to polyurethane organosilicate nanocomposites.

The term nanocomposite refers to organic:inorganic composites where one phase, typically the inorganic phase, has dimensions on the nanoscale. Several authors have noted the potential benefit of biomedical application of nanocomposite technology, and have suggested using quaternary ammonium compounds (QAC) as an organic modification to enhance dispersion of nanoparticles within polymer matrices. This study aimed to examine fibroblast responses in vitro to a range of nanocomposites using different organic modifiers. Composite materials were prepared from a polyether urethane (PEU) and various unmodified and organically modified montmorillonite (MMT) nanoparticles. QAC and amino undecanoic acid (AUA) modified-MMT were added to PEU at loadings ranging from approximately 1 to 15 wt %. Composites with organically modified QAC and AUA particles displayed partially exfoliated and intercalated silicate morphology, respectively. Nanocomposites showed increases in ultimate tensile properties for materials with lower QACMMT loadings. However QAC was shown to significantly inhibit cell growth following release from PEU-QACMMT under extraction conditions mimicking those of the physiological environment. Materials containing silicate modified using AUA were cytocompatible. The results of this study suggest that QAC may be unsuitable as organic modifiers for nanoparticles destined for biomedical use. Alternative modifiers based on AUA confer equivalent dispersion and are of low toxicity.