New prebiotic chemistry inspired filter media for stormwater/greywater disinfection.

Greywater and stormwater have received significant attention due to increasing water scarcity. Passive filtration such as biofiltration has been a popular treatment method with its low energy input and environmental friendliness. However, pathogen removal capacity needs improvement to achieve safe water quality. In this study, a prebiotic chemistry inspired copolymer based on aminomalononitrile and 3,4,5-trihydroxybenzaldehyde (AMNT30) was introduced to develop antimicrobial media for passive filtration. The AMNT30 polymer provided an adhesive coating on zeolite substrates following a spontaneous polymerisation process at room temperature. AMNT30 coated media were investigated for metal loading capacity, surface morphology, E. coli removal and metal leaching after filtration of different water sources (i.e. stormwater, greywater, and deionised water) at low/high conductivity. The coating enhanced metal ion loading on the surface and demonstrated that >8 log reduction of E. coli can be achieved for silver loaded materials compared to a 1 log reduction for copper loaded materials. The coating also increased the stability of the metals on the media irrespective of inflow characteristics. This study provided the first example using AMNT30 to create antimicrobial water purification media. It is expected that this technology will find applications in the water treatment industry.

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.

Application of layer-by-layer coatings to tissue scaffolds - development of an angiogenic biomaterial.

Tissue engineered materials aimed at wound care typically underperform due to poor engrafting to the wound bed. The need for such materials will continue to intensify as a result of an ageing population and an increase in patients suffering from vascular problems. Here we describe the development of an angiogenic coating strategy employing a combination of plasma phase deposition of acrylic acid and layer-by-layer (LBL) chemistry using polyethyleneimine and poly(acrylic acid) for the immobilization of heparin and Vascular Endothelial Growth Factor (VEGF). The formation of the coating and its ability to immobilize heparin was examined by Quartz Crystal Microbalance with Dissipation. X-ray Photoelectron Spectroscopy (XPS) and Atomic Force Microscopy were used to confirm that these coatings retained a significant amount of heparin on the surface when applied to a flat substrate. The coating strategy was transferred to 2 different tissue scaffold architectures: a commercially available non-biodegradable polypropylene mesh, and a biodegradable electrospun poly(lactic-co-glycolic acid) (PLGA) scaffold. XPS confirmed that the coating was successfully applied to the scaffolds and that a similar amount of heparin was immobilized. In vitro testing showed that while HDMEC readily attached to the PLGA scaffold, they were inhibited from adhering and forming proliferative colonies where heparin alone was attached to the LBL coated PLGA scaffold. However, after dip coating with VEGF, the heparin coated scaffold supported both attachment and colony growth of HDMEC; no such colony formation occurred in the absence of VEGF.

Micropatterning of polymer brushes: grafting from dewetting polymer films for biological applications.

In this novel platform, a micropatterned polymer brush was obtained by grafting poly(poly(ethylene glycol) methyl ether methacrylate) (poly(PEGMA)) from a thin macroinitiator film using atom transfer radical polymerization (ATRP). A pattern of holes was formed in the macroinitiator film by taking advantage of its spontaneous dewetting above the glass transition temperature from a bottom polystyrene film, driven by unfavorable intermolecular forces. Patterning by dewetting can be achieved at length-scales from a few hundred nanometers to several tens of micrometers, by simply thermally annealing the bilayer above the glass transition temperature of the polymer. This approach is substrate-independent, as polymer films can be cast onto surfaces of different size, shape, or material. As a demonstration of its potential, proteins, and individual cells were attached on targeted bioadhesive polystyrene areas of the micropatterns within poly(PEGMA) protein-repellent brushes. We anticipate this approach will be suitable for the patterning of brushes, especially for biomedical applications such as in the study of single cells and of cell cocultures.