Jonathan Yeow

Copper-Mediated Living Radical Polymerization (Atom Transfer Radical Polymerization and Copper(0) Mediated Polymerization): From Fundamentals to Bioapplications

Radical Polymerization and Copper(0) Mediated Polymerization): From Fundamentals to Bioapplications Cyrille Boyer,*,†,‡ Nathaniel Alan Corrigan,‡ Kenward Jung,‡ Diep Nguyen,‡ Thuy-Khanh Nguyen,‡ Nik Nik M. Adnan,†,‡ Susan Oliver,†,‡ Sivaprakash Shanmugam,‡ and Jonathan Yeow†,‡ †Australian Centre for Nanomedicine, and ‡Centre for Advanced Macromolecular Design (CAMD), School of Chemical Engineering, University of New South Wales, Sydney 2052, Australia

Pair correlation microscopy reveals the role of nanoparticle shape in intracellular transport and site of drug release

Nanoparticle size, surface charge and material composition are known to affect the uptake of nanoparticles by cells. However, whether nanoparticle shape affects transport across various barriers inside the cell remains unclear. Here we used pair correlation microscopy to show that polymeric nanoparticles with different shapes but identical surface chemistries moved across the various cellular barriers at different rates, ultimately defining the site of drug release. We measured how micelles, vesicles, rods and worms entered the cell and whether they escaped from the endosomal system and had access to the nucleus via the nuclear pore complex. Rods and worms, but not micelles and vesicles, entered the nucleus by passive diffusion. Improving nuclear access, for example with a nuclear localization signal, resulted in more doxorubicin release inside the nucleus and correlated with greater cytotoxicity. Our results therefore demonstrate that drug delivery across the major cellular barrier, the nuclear envelope, is important for doxorubicin efficiency and can be achieved with appropriately shaped nanoparticles.

An Efficient and Highly Versatile Synthetic Route to Prepare Iron Oxide Nanoparticles/Nanocomposites with Tunable Morphologies

We report a versatile synthetic method for the in situ self-assembly of magnetic-nanoparticle-functionalized polymeric nanomorphologies, including spherical micelles and rod-like and worm-like micelles and vesicles. Poly(oligoethylene glycol methacrylate)-block-(methacrylic acid)-block-poly(styrene) (POEGMA-b-PMAA-b-PST) triblock copolymer chains were simultaneously propagated and self-assembled via a polymerization-induced self-assembly (PISA) approach. Subsequently, the carboxylic acid groups in the copolymers were used to complex an iron ion (Fe(II)/Fe(III)) mixture. Iron oxide nanoparticles were then formed in the central block, within the polymeric nanoparticles, via alkaline coprecipitation of the iron(II) and (III) salts. Nanoparticle morphologies, particle sizes, molecular weights, and chemical structures were then characterized by transmission electron microscopy (TEM), dynamic light scattering (DLS), size exclusion chromatography (SEC), and (1)H NMR measurements. TEM micrographs showed that the average size of the magnetic nanoparticles was ∼7 nm at the hydrophobic/hydrophilic nexus contained within the nanoparticles. In addition, XRD was used to confirm the formation of iron oxide nanoparticles. Importantly, the polymeric nanoparticle morphologies were not affected by the coprecipitation of the magnetic nanoparticles. The hybrid nanoparticles were then evaluated as negative MRI contrast agents, displaying remarkably high transverse relaxivities (r2, greater than 550 mM(-1) s(-1) at 9.4 T); a result, that we hypothesize, ensues from iron oxide nanoparticle clustering at the hydrophobic-hydrophilic interface. This simple synthetic procedure is highly versatile and produces nanocarriers of tunable size and shape with high efficacy as MRI contrast agents and potential utility as theranostic delivery vectors.

Oxygen tolerant photopolymerization for ultralow volumes

A benchtop approach is developed for the synthesis of various polymeric architectures using an aqueous Reversible Addition–Fragmentation chain Transfer (RAFT) photopolymerization technique. Under visible green light irradiation (λ = 530 nm), eosin Y (EY) in the presence of ascorbic acid (AscA) as a reducing agent can initiate RAFT polymerization of a range of monomers (acrylamide, acrylate and methacrylate families) in water. More importantly, this process proceeds rapidly without the need for traditional deoxygenation and thus allows RAFT polymerizations to be performed in ultralow volumes (20 μL). This photopolymerization approach can be applied on a 96-well microtiter plate for the synthesis of a range of homopolymer and diblock copolymers. Furthermore, more complex polymeric architectures such as star polymers (arm first) and polymeric nanoparticles (via a polymerization-induced self-assembly (PISA) approach) were successfully synthesized in low volumes and without prior deoxygenation.

Photoinitiated Polymerization‐Induced Self‐Assembly (Photo‐PISA): New Insights and Opportunities

The polymerization‐induced self‐assembly (PISA) process is a useful synthetic tool for the efficient synthesis of polymeric nanoparticles of different morphologies. Recently, studies on visible light initiated PISA processes have offered a number of key research opportunities that are not readily accessible using traditional thermally initiated systems. For example, visible light mediated PISA (Photo‐PISA) enables a high degree of control over the dispersion polymerization process by manipulation of the wavelength and intensity of incident light. In some cases, the final nanoparticle morphology of a single formulation can be modulated by simple manipulation of these externally controlled parameters. In addition, temporal (and in principle spatial) control over the Photo‐PISA process can be achieved in most cases. Exploitation of the mild room temperature polymerizations conditions can enable the encapsulation of thermally sensitive therapeutics to occur without compromising the polymerization rate and their activities. Finally, the Photo‐PISA process can enable further mechanistic insights into the morphological evolution of nanoparticle formation such as the effects of temperature on the self‐assembly process. The purpose of this mini‐review is therefore to examine some of these recent advances that have been made in Photo‐PISA processes, particularly in light of the specific advantages that may exist in comparison with conventional thermally initiated systems.

Application of oxygen tolerant PET-RAFT to polymerization-induced self-assembly

The presence of oxygen during controlled/living radical polymerization (CLRP) can lead to a number of undesirable behaviours such as long inhibition periods and slow polymerization rates. Photoinduced Electron/Energy Transfer-Reversible Addition–Fragmentation chain Transfer (PET-RAFT) polymerization has been shown to possess tolerance to molecular oxygen in dimethylsulfoxide (DMSO) as solvent. This mechanism of oxygen tolerance is due to the rapid conversion of triplet oxygen into singlet oxygen, and then, its rapid consumption in the presence of the singlet oxygen quencher, dimethylsulfoxide. In this work, we demonstrate that the addition of singlet oxygen quenchers such as 9,10-dimethylanthracene (DMA), ascorbic acid (AscA) and (R)-(+)-limonene (Lim) allows for oxygen tolerant PET-RAFT polymerization to be achieved in a range of other organic solvents and under low energy red light (λmax = 635 nm, 3.0 mW cm−2). In particular, when DMA is used as a quencher, remarkably similar polymerization kinetics are observed regardless of prior deoxygenation. Finally, the ability to perform in situ deoxygenation was exploited to conduct RAFT dispersion polymerization of benzyl methacrylate (BzMa) under ethanolic conditions and without inert gas sparging. Under these conditions, nanoparticles of different morphologies (spheres, worm-like micelles and vesicles) can be synthesized according to a polymerization-induced self-assembly (PISA) approach. This work greatly expands the versatility of PET-RAFT polymerizations by allowing oxygen tolerance to be achieved under a broader range of polymerization conditions.

Polymerization of a Photocleavable Monomer Using Visible Light.

The polymerization of the photocleavable monomer, o-nitrobenzyl methacrylate (NBMA), is investigated using photoinduced electron/energy transfer reversible addition-fragmentation chain transfer polymerization. The polymerizations under visible red (λ max = 635 nm, 0.7 mW cm(-2) ) and yellow (λ max = 560 nm, 9.7 mW cm(-2) ) light are performed and demonstrate rational evidence of a controlled/living radical polymerization process. Well-defined poly(o-nitrobenzyl methacrylate) (PNBMA) homopolymers with good control over the molecular weight and polymer dispersity are successfully synthesized by varying the irradiation time and/or targeted degree of polymerization. Chain extension of a poly(oligo(ethylene glycol) methyl ether methacrylate) macro-chain transfer agent with NBMA is carried out to fabricate photocleavable amphiphilic block copolymers (BCP). Finally, these self-assembled BCP rapidly dissemble under UV light suggesting the photoresponsive character of NBMA is not altered during the polymerization under yellow or red light. Such photoresponsive polymers can be potentially used for the remote-controlled delivery of therapeutic compounds.

An Oxygen‐Tolerant PET‐RAFT Polymerization for Screening Structure–Activity Relationships

The complexity of polymer-protein interactions makes rational design of the best polymer architecture for any given biointerface extremely challenging, and the high throughput synthesis and screening of polymers has emerged as an attractive alternative. A porphyrin-catalysed photoinduced electron/energy transfer-reversible addition-fragmentation chain-transfer (PET-RAFT) polymerisation was adapted to enable high throughput synthesis of complex polymer architectures in dimethyl sulfoxide (DMSO) on low-volume well plates in the presence of air. The polymerisation system shows remarkable oxygen tolerance, and excellent control of functional 3- and 4-arm star polymers. We then apply this method to investigate the effect of polymer structure on protein binding, in this case to the lectin concanavalin A (ConA). Such an approach could be applied to screen the structure-activity relationships for any number of polymer-protein interactions.

The effects of polymer topology and chain length on the antimicrobial activity and hemocompatibility of amphiphilic ternary copolymers

Investigation into the macromolecular structure–activity relationship of synthetic antimicrobial polymers has been gaining scientific interest due to the possibility of discovering new alternatives for combating the increase of multidrug resistance in bacteria. Recently, we reported the development of new antimicrobial polymers in the form of amphiphilic ternary copolymers that consist of low-fouling (oligoethylene glycol), cationic and hydrophobic side chains. The combination of these three main functional groups is crucial in endowing the polymers with high antimicrobial potency against Gram-negative pathogens and low cytotoxicity. Following on from our previous study, we herein present a systematic assessment on the effects of the polymer chain length and architecture (i.e., random vs. block copolymers and linear vs. hyperbranched) on the antimicrobial activity and hemocompatibility of antimicrobial ternary copolymers. The polymer chain length in random copolymers slightly affects the antimicrobial activity where longer chains are marginally more bacteriostatic against Pseudomonas aeruginosa and Escherichia coli. In terms of hemocompatibility, polymers with shorter chains are more prone to hemagglutination. Interestingly, when the hydrophilic and hydrophobic segments are separated into diblock copolymers, the antimicrobial activity is lost, possibly due to the stable core–shell architecture. The hyperbranched structure which consists of 2-ethylhexyl groups as hydrophobic side-chains yields the best overall biological properties, having similar antimicrobial activity (MIC = 64 μg mL−1) and >4-fold increase in HC50 compared to the linear random copolymers (HC50 > 10 000 μg mL−1) with no hemagglutination. The hyperbranched polymers are also bactericidal and kill ≥99% and 90% of planktonic and biofilm Pseudomonas aeruginosa, respectively. This study thus highlights the importance of determining macromolecular structural aspects that govern the biological activity of antimicrobial polymers.

Facile Synthesis of Worm-like Micelles by Visible Light Mediated Dispersion Polymerization Using Photoredox Catalyst

Presented herein is a protocol for the facile synthesis of worm-like micelles by visible light mediated dispersion polymerization. This approach begins with the synthesis of a hydrophilic poly(oligo(ethylene glycol) methyl ether methacrylate) (POEGMA) homopolymer using reversible addition-fragmentation chain-transfer (RAFT) polymerization. Under mild visible light irradiation (λ = 460 nm, 0.7 mW/cm(2)), this macro-chain transfer agent (macro-CTA) in the presence of a ruthenium based photoredox catalyst, Ru(bpy)3Cl2 can be chain extended with a second monomer to form a well-defined block copolymer in a process known as Photoinduced Electron Transfer RAFT (PET-RAFT). When PET-RAFT is used to chain extend POEGMA with benzyl methacrylate (BzMA) in ethanol (EtOH), polymeric nanoparticles with different morphologies are formed in situ according to a polymerization-induced self-assembly (PISA) mechanism. Self-assembly into nanoparticles presenting POEGMA chains at the corona and poly(benzyl methacrylate) (PBzMA) chains in the core occurs in situ due to the growing insolubility of the PBzMA block in ethanol. Interestingly, the formation of highly pure worm-like micelles can be readily monitored by observing the onset of a highly viscous gel in situ due to nanoparticle entanglements occurring during the polymerization. This process thereby allows for a more reproducible synthesis of worm-like micelles simply by monitoring the solution viscosity during the course of the polymerization. In addition, the light stimulus can be intermittently applied in an ON/OFF manner demonstrating temporal control over the nanoparticle morphology.