Mihee Kim

pH-dependent structures of ferritin and apoferritin in solution: disassembly and reassembly.

The pH-dependent structures of the ferritin shell (apoferritin, 24-mer) and the ferrihydrite core, under physiological conditions that permit enzymatic activity, were investigated by synchrotron small-angle X-ray scattering (SAXS). The solution structure of apoferritin was found to be nearly identical to the crystal structure. The shell thickness and hollow core volumes were estimated. The intact hollow spherical apoferritin was stable over a wide pH range, 3.40-10.0, and the ferrihydrite core was stable over the pH range 2.10-10.0. The apoferritin subunits underwent aggregation below pH 0.80, whereas the ferrihydrite cores aggregated below pH 2.10 as a result of the disassembly of the ferritin shell under the strongly acidic conditions. As the pH decreased from 3.40 to 0.80, apoferritin underwent stepwise disassembly by first forming a hollow sphere with two holes, then a headset-shaped structure, and, finally, rodlike oligomers. As the pH was increased from pH 1.96, the disassembled rodlike oligomers recovered only to the headset-shaped structure, and the disassembled headset-shaped intermediates recovered only to the hollow spherical structure with two hole defects. The apoferritin hole defects that formed during the disassembly process did not heal as the pH was increased to neutral or slightly basic conditions. The pH-induced apoferritin disassembly and reassembly processes were not fully reversible, although they were pseudoreversible over a limited pH range, between 10.0 and 2.66.

Well-defined DNA-mimic brush polymers bearing adenine moieties: synthesis, layer-by-layer self-assembly, and biocompatibility.

Two new DNA-mimicking brush polymers were synthesized: poly[oxy(11-(3-(9-adeninyl)propionato)-undecanyl-1-thiomethyl)ethylene] (PECH-AP) and poly[oxy(11-(5-(9-adenylethyloxy)-4-oxopentanoato)undecanyl-1-thiomethyl)ethylene] (PECH-AS). These polymers were found to be thermally stable up to 220 °C and could be applied easily by conventional coating processes to produce good quality films. Interestingly, both brush polymers formed molecular multibilayer structures to provide an adenine-rich surface. Despite the structural similarities, PECH-AS surprisingly exhibited higher hydrophilicity and better water sorption properties than PECH-AP. These differences were attributed to the chemical structures in the bristles of the polymers. The adenine-rich surfaces of the polymer films demonstrated selective protein adsorption, suppressed bacterial adherence, facilitated HEp-2 cell adhesion, and exhibited good biocompatibility in mice. However, the high hydrophilicity and good water sorption characteristics of the PECH-AS film suggest that this brush polymer is better suited to applications requiring good biocompatibility and reduced chance of bacterial infection compared with the PECH-AP film.

Molecular layer-by-layer self-assembly and mercury sensing characteristics of novel brush polymers bearing thymine moieties.

Two new brush polyoxyethylenes bearing thymine moieties at the bristle ends have been synthesized as model polymers in which the chemical loading of the thymine functional group into the polymer is maximized: poly(oxy(11-thyminoacetyloxyundecylthiomethyl)ethylene) (PECH(S)-T) and poly(oxy(11-thyminoacetyloxyundecylsulfonylmethyl)ethylene) (PECH(SO(2))-T). These brush polymers are thermally stable up to around 225 °C, and their glass transitions occur in the range 23-27 °C, but they have significantly different properties despite the similarity of their chemical structures. In particular, PECH(SO(2))-T films exhibit better performance in sensing mercury ions than PECH(S)-T films. These differences were found to originate in the differences between their morphological structures. The PECH(SO(2))-T film has a multi-bilayer structure without interdigitation, in which the layers stack along the out-of-plane of the film and provide a thymine-rich surface. In contrast, the PECH(S)-T film is amorphous with a relatively low population of thymine moieties at the surface. This study demonstrated that a thymine-rich surface is required for recyclable thymine-based polymers to provide highly improved sensitivity and selectivity as well as full reversibility in the sensing of mercury ions. A thymine-rich surface can be achieved with a brush polymer bearing thymine moieties that can self-assemble into a multi-bilayer structure. Because of the thymine-rich surface, the PECH(SO(2))-T thin films even in only 6 nm thickness demonstrate the detection of mercury ions in aqueous solutions with a detection limit of 10(-6) M.

Bacterial adherence on self-assembled films of brush polymers bearing zwitterionic sulfobetaine moieties

In this study we synthesized a series of well-defined brush polymers, poly(oxy(11-(3-sulfonylpropyltrimethylglycinyl)undecylesterthiomethyl)ethylene-co-oxy(n-dodecylthiomethyl)-ethylene)s (PECH-DMAPSm, where m is the mol% of DMAPS (sulfobetaine) end group). The thermal properties and phase transitions of these polymers were investigated. The polymers were thermally stable up to 185 °C. The polymers were found to form favorably into multi-bilayer structures, always providing hydrophilic, zwitterionic sulfobetaine end groups at the film surface. For the films, water sorption behavior was examined. In addition, surface energy components were determined for the polymer films and the bacterial cells deposited on cellulose acetate membranes. The brush polymer films were found to suppress bacterial adherence significantly. An understanding of the suppression of bacterial adherence was attempted in terms of surface energies and thermodynamics. The results collectively indicate that the sulfobetaine-containing brush polymers are suitable for use in biomedical applications that require the reduced possibility of post-operative infection.

New self-assembled brush glycopolymers: Synthesis, structure and properties

A new series of chemically well-defined brush glycopolymers consisting of a polyoxyethylene backbone and bristles bearing glycosyl and methyl end groups was synthesized with various compositions. The glycopolymers were thermally stable up 200 °C and were soluble in a variety of common solvents. The brush polymer films formed multibilayer structures, the layers of which were stacked along the direction normal to the film plane so as to display a glycosyl group-rich surface or a methyl group-rich surface or their mixture, depending on the bristle end group composition. The multibilayer structures were stabilized by the self-assembly of the bristles via lateral packing. The glycosyl-rich surface played a critical role in enhancing the surface hydrophilicity and water sorption to a certain level; thus, the glycopolymer films easily formed a hydration layer to a certain depth on the film surface. The hydrophilic surfaces and hydration layer efficiently prevented protein adsorption onto the brush glycopolymers and suppressed bacterial adherence while promoting mammalian cell adhesion and displaying excellent biocompatibility in an in vivo mouse study.

Biocompatible characteristics of sulfobetaine-containing brush polymers

AbstractA series of well-defined brush polymers, poly(oxy(11-(3-sulfonylpropyltrimethyl-glycinyl)undecylesterthiomethyl) ethylene-co-oxy(n-dodecylthio-methyl)ethylene)s (PECH-DMAPSm, where m is the mol% of the DMAPS [sulfobetaine] end group) were synthesized. The thermal properties and phase transitions of these polymers were investigated. The polymers were thermally stable up to 185 °C and were found to form favorably into multibilayer structures, always providing hydrophilic, zwitterionic sulfobetaine end groups at the film surface. Because of the presence of these sulfobetaine groups at the surface, the polymer films promoted HEp-2 cell adhesion and revealed biocompatibility in mice but significantly suppressed protein adsorption. These results collectively indicate that the sulfobetaine-containing brush polymers are suitable for use in biomedical applications, including medical devices and biosensors that require biocompatibility.

Chemical End Group Modified Diblock Copolymers Elucidate Anchor and Chain Mechanism of Membrane Stabilization

Block copolymers can be synthesized in an array of architectures and compositions to yield diverse chemical properties. The triblock copolymer Poloxamer 188 (P188), the family archetype, consisting of a hydrophobic poly(propylene oxide) core flanked by hydrophilic poly(ethylene oxide) chains, can stabilize cellular membranes during stress. However, little is known regarding the molecular basis of membrane interaction by copolymers in living organisms. By leveraging diblock architectural design, discrete end-group chemistry modifications can be tested. Here we show evidence of an anchor and chain mechanism of interaction wherein titrating poly(propylene oxide) block end group hydrophobicity directly dictates membrane interaction and stabilization. These findings, obtained in cells and animals in vivo, together with molecular dynamics simulations, provide new insights into copolymer-membrane interactions and establish the diblock copolymer molecular architecture as a valuable platform to inform copolymer-biological membrane interactions. These results have implications for membrane stabilizers in muscular dystrophy and for other biological applications involving damaged cell membranes.

PEO–PPO Diblock Copolymers Protect Myoblasts from Hypo-Osmotic Stress In Vitro Dependent on Copolymer Size, Composition, and Architecture

Poloxamer 188, a triblock copolymer of poly(ethylene oxide) (PEO) and poly(propylene oxide) (PPO), protects cellular membranes from various stresses. Though numerous block copolymer variants exist, evaluation of alternative architecture, composition, and size has been minimal. Herein, cultured murine myoblasts are exposed to the stresses of hypotonic shock and isotonic recovery, and membrane integrity was evaluated by quantifying release of lactate dehydrogenase. Comparative evaluation of a systematic set of PEO-PPO diblock and PEO-PPO-PEO triblock copolymers demonstrates that the diblock architecture can be protective in vitro. Short PPO blocks hinder protection with >9 PPO units needed for protection at 150 μM and >16 units needed at 14 μM. Addition of a tert-butyl end group enhances protection at reduced concentration. When the end group and PPO length are fixed, increasing the PEO length improves protection. This systematic evaluation establishes a new in vitro screening tool for evaluating membrane-sealing amphiphiles and provides mechanistic insight to guide future copolymer design for membrane stabilization in vivo.

Surface Plasmon Resonance Study of the Binding of PEO–PPO–PEO Triblock Copolymer and PEO Homopolymer to Supported Lipid Bilayers

Poloxamer 188 (P188), a poly(ethylene oxide)- b-poly(propylene oxide)- b-poly(ethylene oxide) triblock copolymer, protects cell membranes against various external stresses, whereas poly(ethylene oxide) (PEO; 8600 g/mol) homopolymer lacks protection efficacy. As part of a comprehensive effort to elucidate the protection mechanism, we used surface plasmon resonance (SPR) to obtain direct evidence of binding of the polymers onto supported lipid bilayers. Binding kinetics and coverage of P188 and PEO were examined and compared. Most notably, PEO exhibited membrane association comparable to that of P188, evidenced by comparable association rate constants and coverage. This result highlights the need for additional mechanistic understanding beyond simple membrane association to explain the differential efficacy of P188 in therapeutic applications.

2.16 – Reflectivity, GI-SAS and GI-Diffraction: X-Ray

One of the features of X-rays is the penetration power into matter, revealing the invisible interior of complex objects. Moreover, X-rays are a critical tool to investigate material structures and properties because of the electromagnetic radiation that is able to interact with matter having an electromagnetic nature. In particular, X-ray reflectivity (XR) and grazing incidence X-ray scattering (GIXS) are powerful, nondestructive techniques for examining the structure and properties of materials including various kinds of polymers and their nanostructures and nanosize specimens. These techniques have become more powerful for understanding nanostructures and objects in nanosize by using synchrotron radiation sources. Their fundamental theories and applications in polymer science are given and discussed.