Thomas R. Congdon

Thermoresponsive, well-defined, poly(vinyl alcohol) co-polymers

Thermoresponsive polymers have attracted huge interest as adaptable biomaterials based on their reversible solubility behaviour which can be exploited for controlled drug delivery or cellular uptake. The most famous and successful of these is poly(ethylene glycol) (PEG), but the thermal transition temperatures that are practically accessible are not physiologically useful. There are some notable examples of synthetic, responsive, polymers that are highly tunable over a physiologically relevant range, but there is still a need for these to be clinically validated in terms of toxicology and immunogenity for in vivo usage, in addition to their widely used in vitro applications. Poly(vinyl alcohol), PVA, is an appealing biocompatible polymer which is already used for a huge range of biomedical applications. Here, PVA is shown to be a highly tunable, thermoresponsive polymer scaffold. RAFT/MADIX polymerization is used to obtain a library of well-defined polymers between 8 and 50 kDa. Selective alkanoylation of the obtained PVA enabled the effect of side-chains, end-groups and molecular weight on the observable transition temperatures to be studied by turbidimetry. It was found that increasingly hydrophobic side chains (acetyl, propanoyl, butanoyl), or increasing their density led to corresponding decreases in cloud point. PVA with just 10 mol% butanoylation was shown to have a thermal transition temperature close to physiological temperatures (37 °C), compared to 70 mol% for acetylation, with temperatures in between accessible by controlling both the relative degree of functionalization, or by altering the chain length. Finally, a secondary response to esterase enzymes was demonstrated as a route to ‘turn off’ the responsive behaviour on demand. This study suggests that PVA-derived polymers may be a useful platform for responsive biomaterials.

Probing the biomimetic ice nucleation inhibition activity of poly(vinyl alcohol) and comparison to synthetic and biological polymers

Nature has evolved many elegant solutions to enable life to flourish at low temperatures by either allowing (tolerance) or preventing (avoidance) ice formation. These processes are typically controlled by ice nucleating proteins or antifreeze proteins, which act to either promote nucleation, prevent nucleation or inhibit ice growth depending on the specific need, respectively. These proteins can be expensive and their mechanisms of action are not understood, limiting their translation, especially into biomedical cryopreservation applications. Here well-defined poly(vinyl alcohol), synthesized by RAFT/MADIX polymerization, is investigated for its ice nucleation inhibition (INI) activity, in contrast to its established ice growth inhibitory properties and compared to other synthetic polymers. It is shown that ice nucleation inhibition activity of PVA has a strong molecular weight dependence; polymers with a degree of polymerization below 200 being an effective inhibitor at just 1 mg.mL–1. Other synthetic and natural polymers, both with and without hydroxyl-functional side chains, showed negligible activity, highlighting the unique ice/water interacting properties of PVA. These findings both aid our understanding of ice nucleation but demonstrate the potential of engineering synthetic polymers as new biomimetics to control ice formation/growth processes

Influence of Block Copolymerization on the Antifreeze Protein Mimetic Ice Recrystallization Inhibition Activity of Poly(vinyl alcohol)

Antifreeze (glyco) proteins are produced by many cold-acclimatized species to enable them to survive subzero temperatures. These proteins have multiple macroscopic effects on ice crystal growth which makes them appealing for low-temperature applications—from cellular cryopreservation to food storage. Poly(vinyl alcohol) has remarkable ice recrystallization inhibition activity, but its mode of action is uncertain as is the extent at which it can be incorporated into other high-order structures. Here the synthesis and characterization of well-defined block copolymers containing poly(vinyl alcohol) and poly(vinylpyrrolidone) by RAFT/MADIX polymerization is reported, as new antifreeze protein mimetics. The effect of adding a large second hydrophilic block is studied across a range of compositions, and it is found to be a passive component in ice recrystallization inhibition assays, enabling retention of all activity. In the extreme case, a block copolymer with only 10% poly(vinyl alcohol) was found to retain all activity, where statistical copolymers of PVA lose all activity with very minor changes to composition. These findings present a new method to increase the complexity of antifreeze protein mimetic materials, while retaining activity, and also to help understand the underlying mechanisms of action.

Gold Nanoparticle Aggregation as a Probe of Antifreeze (Glyco) Protein-Inspired Ice Recrystallization Inhibition and Identification of New IRI Active Macromolecules.

Antifreeze (glyco)proteins are found in polar fish species and act to slow the rate of growth of ice crystals; a property known as ice recrystallization inhibition. The ability to slow ice growth is of huge technological importance especially in the cryopreservation of donor cells and tissue, but native antifreeze proteins are often not suitable, nor easily available. Therefore, the search for new materials that mimic this function is important, but currently limited by the low-throughout assays associated with the antifreeze properties. Here 30 nm gold nanoparticles are demonstrated to be useful colorimetric probes for ice recrystallization inhibition, giving a visible optical response and is compatible with 96 well plates for high-throughout studies. This method is faster, requires less infrastructure, and has easier interpretation than the currently used ‘splat’ methods. Using this method, a series of serum proteins were identified to have weak, but specific ice recrystallization inhibition activity, which was removed upon denaturation. It is hoped that high-throughput tools such as this will accelerate the discovery of new antifreeze mimics.

Synthesis of polymeric microcapsules by interfacial-suspension cationic photopolymerisation of divinyl ether monomer in aqueous suspension

Polymeric microcapusles have been synthesised with a markedly more hydrophillic monomer than previously reported, triethylene glycol divinyl ether, using cationic photopolymerisation in an aqueous environment. Characterisation by NMR and SEM show that the particles are formed with low dispersity with a size of approximately 1 μm in spite of the expected inhibition in aqueous conditions. Furthermore, supercritical carbon dioxide has been used to generate silver nanoparticles which distribute throughout the shell of the microcapsules further illustrating the structure of these capsules with characterisation by TEM and SAXS.