Mitsuhiro Ebara

Novel bifunctional polymer with reactivity and temperature sensitivity

To introduce reactive groups into temperature-responsive polymeric chains of poly(N-isopropylacrylamide) (PIPAAm), IPAAm is copolymerized with other monomer such as acrylic acid (AAc). IPAAm homopolymer exhibited temperature-responsive properties and phase transition at 32°C, however, the lower critical solution temperature (LCST) of the IPAAm-AAc copolymer shifts to a higher temperature and the phase transition becomes insensitive with increasing AAc content. To achieve a useful bifunctional copolymer containing both reactivity and temperature-sensitivity, we assumed that the homopolymer-like structure in the polymer chain would be required to maintain a sensitive temperature response with functional groups. Therefore, we designed a reactive monomer, 2-carboxyisopropylacrylamide (CIPAAm), and investigated its copolymerization with IPAAm. The important characteristic of the poly(IPAAm-co-CIPAAm) structure is that it was composed of the same polymer backbone and isopropylamide groups and some additional carboxyl groups. The transmittance measurement of the polymer aqueous solution revealed that phase transition of IPAAm-co-CIPAAm random copolymer occurred within a very narrow temperature range in pH 6.4, 7.4, and also even 9.0 phosphate buffered solution. These profiles were almost same as that of IPAAm homopolymer. While, under the same conditions, phase transition properties of poly(IPAAm-co-AAc)s solution were considerably influenced by small AAc content. We succeeded with the preparation of bifunctional polymer that possessed reactive functional groups and very sensitive response to temperature change.

Shape-Memory Surface with Dynamically Tunable Nano-Geometry Activated by Body Heat

Shape-memory surfaces with on-demand, tunable nanopatterns are developed to observe time dependent changes in cell alignment using temperature-responsive poly(ϵ-caprolactone) (PCL) films. Temporary grooved nanopatterns are easily programmed on the films and triggered to transition quickly to permanent surface patterns by the application of body heat. A time-dependent cytoskeleton remodeling is also observed under biologically relevant conditions.

Bioinspired nanoarchitectonics as emerging drug delivery systems

We propose an important paradigm shift in the preparation of functional materials with well-designed nanostructures, from the nanotechnological to the nanoarchitectonic approach. Nanoarchitectonics is a methodology for arranging nanoscale structural units in the required configurations for new functional materials by the sophisticated combination and harmonization of several processes including atom/molecule manipulation, chemical nanomanipulation, field-induced materials control and controlled supramolecular self-assembly. In particular, nanoarchitectonics bears features of nanoscale phenomena including their flexibility and uncertainty of structure due to the unavoidable influence of thermal fluctuation. It shares characteristics with structural constructions in biological systems and could become a powerful bioinspired approach for materials science. Here, we focus on examples involving drug delivery functions due to these promising applications of bioinspired materials research. We commence with a discussion of recent developments involving assemblies of small amphiphilic molecules, polymer micelles and molecular conjugates and follow this with examples of challenging concepts including inorganic nanostructure design for drug delivery and mechanically controlled drug release. The new concept of bioinspired nanoarchitectonics could significantly expand the possibilities of systems design for drug delivery.

Photo-switchable control of pH-responsive actuators via pH jump reaction

We propose a new approach to fabricate reversible self-bending actuators utilizing a photo-triggered pH jump reaction. A photo-initiated proton-releasing agent of o-nitrobenzaldehyde (NBA) was successfully integrated into bilayer hydrogels composed of a polyacid layer, poly(N-isopropylacrylamide-co-2-carboxyisopropylacrylamide) (P(NIPAAm-co-CIPAAm)) and a polybase layer, poly(N-isopropylacrylamide-co-N,N′-dimethylaminopropylacylamide) (P(NIPAAm-co-DMAPAAm)), where the adhesion of both layers was achieved via electrophoresis of semi-interpenetrating polyelectrolyte chains. The NBA-integrated bilayer gels demonstrated quick proton release upon UV irradiation, allowing the pH within the gel to decrease below the volume phase transition pH in 30 seconds. By controlling the NBA concentration and the gel thickness, the degrees and the kinetics of bending were easily controlled. Reversible bending was also studied with respect to the NBA concentration in response to ‘on–off’ UV irradiation. Additionally, self-bending of the non-UV irradiated region of the gel was also achieved because the generated protons gradually diffused toward the non-irradiated region. The proposed system can be potentially applied in the fields of mechanical actuators, controlled encapsulation and drug release, robotics and microfluidic technologies because control over autonomous motion by both physical and chemical signals is essential as a programmable system for real biomedical and nano-technological applications.

A Smart Nanofiber Web That Captures and Releases Cells

Caught in a web: Photo-cross-linkable temperature-responsive polymer-based nanofiber webs have been synthesized that have the ability to capture, encapsulate, and release cells by dynamically transforming the fibrous structure into hydrogel-like structures by wrapping, swelling, and deswelling processes in response to external temperature changes (see picture).

Hippo pathway effectors control cardiac progenitor cell fate by acting as dynamic sensors of substrate mechanics and nanostructure

Stem cell responsiveness to extracellular matrix (ECM) composition and mechanical cues has been the subject of a number of investigations so far, yet the molecular mechanisms underlying stem cell mechano-biology still need full clarification. Here we demonstrate that the paralog proteins YAP and TAZ exert a crucial role in adult cardiac progenitor cell mechano-sensing and fate decision. Cardiac progenitors respond to dynamic modifications in substrate rigidity and nanopattern by promptly changing YAP/TAZ intracellular localization. We identify a novel activity of YAP and TAZ in the regulation of tubulogenesis in 3D environments and highlight a role for YAP/TAZ in cardiac progenitor proliferation and differentiation. Furthermore, we show that YAP/TAZ expression is triggered in the heart cells located at the infarct border zone. Our results suggest a fundamental role for the YAP/TAZ axis in the response of resident progenitor cells to the modifications in microenvironment nanostructure and mechanics, thereby contributing to the maintenance of myocardial homeostasis in the adult heart. These proteins are indicated as potential targets to control cardiac progenitor cell fate by materials design.

Fabrication of doubly responsive polymer functionalized silica nanoparticles via a simple thiol–ene click chemistry

Temperature and pH responsive silica nanoparticles were easily prepared by a simple thiol–ene click chemistry. Well-defined poly(2-(diethylamino)ethyl methacrylate) (PDEAEMA) and poly(N-isopropylacrylamide) (PNIPAAm) were first synthesized by a reversible addition–fragmentation chain transfer (RAFT) process. The polymers were then reduced to generate a thiol group at the chain end to react with vinyl groups on the surface of silica nanoparticles via a simple ‘click’ reaction. The ratio of the PDEAEMA and PNIPAAm segments on the silica nanoparticles surface was controlled by adjusting the feed weight ratios of the polymers in the reaction solution. The surface coated silica nanoparticles were characterized by a range of techniques such as thermogravimetric analysis (TGA), Fourier transform infrared (FT-IR) spectroscopy, transmission electron microscopy (TEM), and dynamic light scattering (DLS). The functionalized silica nanoparticles showed both pH and temperature responsive behavior and the solution properties were dependent on the ratio of the two polymers on the surface.

Temperature-responsive electrospun nanofibers for ‘on–off’ switchable release of dextran

Abstract We propose a new type of ‘smart’ nanofiber (NF) with dynamically and reversibly tunable properties for the ‘on–off’ controlled release of the polysaccharide dextran. The fibers are produced by electrospinning copolymers of N-isopropylacrylamide (NIPAAm) and N-hydroxymethylacrylamide (HMAAm). The OH groups of HMAAm are subsequently crosslinked by thermal curing. The copolymers were successfully fabricated into a well-defined nanofibrous structure with a diameter of about 600–700 nm, and the fibers preserved their morphology even after thermal curing. The resulting crosslinked NFs showed rapid and reversible volume changes in aqueous media in response to cycles of temperature alternation. The fibrous morphology was maintained for the crosslinked NFs even after the cycles of temperature alternation, while non-crosslinked NFs collapsed and dispersed quickly in the aqueous solution. Dextran-containing NFs were prepared by electrospinning the copolymers blended with fluorescein isothiocyanate (FITC)-dextran, and the ‘on–off’ switchable release of FITC-dextran from the crosslinked NFs was observed. Almost all the FITC-dextran was released from the NFs after six heating cycles, whereas only a negligible amount of FITC-dextran was evolved during the cooling process. The reported incorporation of smart properties into NFs takes advantage of their extremely large surface area and porosity and is expected to provide a simple platform for on–off drug delivery.

Fabrication of zeolite–polymer composite nanofibers for removal of uremic toxins from kidney failure patients

There is a need to develop a simple, cheap, and accessible method of treating patients with kidney failure, especially in resource-limited environments such as disaster areas and the developing world due to the inaccessibility of conventional hemodialysis treatments. In this study, we develop a zeolite-polymer composite nanofiber mesh to remove uremic toxins for blood purification. The nanofiber is composed of blood compatible poly(ethylene-co-vinyl alcohol) (EVOH) as the primary matrix polymer and zeolites which are capable of selectively adsorbing uremic toxins such as creatinine. The composite fiber meshes were produced by a cost-effective electrospinning method: electrospinning composite solutions of EVOH and zeolites. Scanning electron microscope (SEM) images revealed that the 7 w/v% EVOH solution produced non-woven fibers with a continuous and smooth morphology. The SEM also showed that over 90% of zeolites in the solution were successfully incorporated into the EVOH nanofibers. Although the barrier properties of the EVOH matrix lowered the creatinine adsorption capacity of the zeolites in the fiber when compared with adsorption to free zeolites, their adsorption capacity was still 67% of the free zeolites. The proposed composite fibers have the potential to be utilized as a new approach to removing nitrogenous waste products from the bloodstream without the requirement of specialized equipment.

Novel temperature-responsive polymer brushes with carbohydrate residues facilitate selective adhesion and collection of hepatocytes

Abstract Temperature-responsive glycopolymer brushes were designed to investigate the effects of grafting architectures of the copolymers on the selective adhesion and collection of hypatocytes. Homo, random and block sequences of N-isopropylacrylamide and 2-lactobionamidoethyl methacrylate were grafted on glass substrates via surface-initiated atom transfer radical polymerization. The galactose/lactose-specific lectin RCA120 and HepG2 cells were used to test for specific recognition of the polymer brushes containing galactose residues over the lower critical solution temperatures (LCSTs). RCA120 showed a specific binding to the brush surfaces at 37 °C. These brush surfaces also facilitated the adhesion of HepG2 cells at 37 °C under nonserum conditions, whereas no adhesion was observed for NIH-3T3 fibroblasts. When the temperature was decreased to 25 °C, almost all the HepG2 cells detached from the block copolymer brush, whereas the random copolymer brush did not release the cells. The difference in releasing kinetics of cells from the surfaces with different grafting architectures can be explained by the correlated effects of significant changes in LCST, mobility, hydrophilicity and mechanical properties of the grafted polymer chains. These findings are important for designing ‘on–off’ cell capture/release substrates for various biomedical applications such as selective cell separation.