Bioinspired Control of Calcium Phosphate Liesegang Patterns Using Anionic Polyelectrolytes.

The Liesegang phenomenon is a spontaneous pattern formation, which is a periodic distribution of the precipitate discovered in diffusion-limited systems. Over the past century, it has been experimentally attempted to control the periodicity of patterns and structures of precipitates by varying the concentration of the hydrogel or electrolytes, adding organic or inorganic impurities, and applying an electric or pH field. In this work, the periodic patterns of calcium phosphate were manipulated with an anionic macromolecular additive inspired by bone mineralization in which various noncollagenous proteins are involved in the formation of a polymer-induced liquid precursor. The periodic patterns were systematically controlled by adjusting the amount of poly(acrylic acid), and they were numerically simulated by adjusting the threshold concentration of nucleation. The change of the pattern is explained by improved stability and directional diffusion of the intermediate.

Cytocompatible Polymer Grafting from Individual Living Cells by Atom-Transfer Radical Polymerization.

A cytocompatible method of surface-initiated, activator regenerated by electron transfer, atom transfer radical polymerization (SI-ARGET ATRP) is developed for engineering cell surfaces with synthetic polymers. Dopamine-based ATRP initiators are used for both introducing the ATRP initiator onto chemically complex cell surfaces uniformly (by the material-independent coating property of polydopamine) and protecting the cells from radical attack during polymerization (by the radical-scavenging property of polydopamine). Synthetic polymers are grafted onto the surface of individual yeast cells without significant loss of cell viability, and the uniform and dense grafting is confirmed by various characterization methods including agglutination assay and cell-division studies. This work will provide a strategic approach to the generation of living cell-polymer hybrid structures and open the door to their application in multitude of areas, such as sensor technology, catalysis, theranostics, and cell therapy.

Effects of spring-loaded crutches on gastrocnemius activity and upward displacement of the body during gait.

[Purpose] This study investigated the effects of spring-loaded crutches on gastrocnemius muscle activity and upper body displacement in the sagittal plane during gait. [Subjects and Methods] The study involved 12 healthy males. All subjects performed crutch gait by using spring-loaded crutches and axillary crutches. During this gait, the gastrocnemius muscle activity was measured using a wireless electromyography system, and upward displacement of the body was measured using a three-dimensional motion analysis system. [Results] The gastrocnemius activity was significantly lower but upward displacement of the body was significantly greater with the spring-loaded crutches than with axillary crutches. [Conclusion] Spring-loaded crutches allow efficient crutch gait and involve less effort from the gastrocnemius muscle.

Formation of Silica/Graphene Oxide Hybrid Nano Films by Layer-by-Layer Self-Assembly and Biomimetic Silicification.

Silica/graphene oxide hybrid thin films were formed by layer-by-layer self-assembly and biomimetic silicification, and the thickness and structure of hybrid thin films were finely controlled at the nanometer scale, by tuning number of the layer-by-layer process. The physical properties of thin films were characterized by infrared spectroscopy, atomic force microscopy, and scanning electron microscopy. In addition, silica/graphene oxide hybrid thin films were successfully utilized for cell culture platforms.

Cytoprotective silica coating of individual mammalian cells through bioinspired silicification.

The cytoprotective coating of physicochemically labile mammalian cells with a durable material has potential applications in cell-based sensors, cell therapy, and regenerative medicine, as well as providing a platform for fundamental single-cell studies in cell biology. In this work, HeLa cells in suspension were individually coated with silica in a cytocompatible fashion through bioinspired silicification. The silica coating greatly enhanced the resistance of the HeLa cells to enzymatic attack by trypsin and the toxic compound poly(allylamine hydrochloride), while suppressing cell division in a controlled fashion. This bioinspired cytocompatible strategy for single-cell coating was also applied to NIH 3T3 fibroblasts and Jurkat cells.

A cytoprotective and degradable metal-polyphenol nanoshell for single-cell encapsulation.

Single-cell encapsulation promises the cytoprotection of the encased cells against lethal stressors, reminiscent of the sporulation process in nature. However, the development of a cytocompatible method for chemically mimicking the germination process (i.e., shell degradation on-demand) has been elusive, despite the shell degradation being pivotal for the practical use of functional cells as well as for single cell-based biology. We report that an artificial shell, composed of tannic acid (TA) and Fe(III) , on individual Saccharomyces cerevisiae controllably degrades on-demand, while protecting the yeast from multiple external aggressors, including UV-C irradiation, lytic enzymes, and silver nanoparticles. Cell division is suppressed by the TA-Fe(III) shell, but restored fully upon shell degradation. The formation of a TA-Fe(III) shell would provide a versatile tool for achieving the chemical version of "sporulation and germination".

Artificial spores: cytocompatible encapsulation of individual living cells within thin, tough artificial shells.

Cells are encapsulated individually within thin and tough shells in a cytocompatible way, by mimicking the structure of bacterial endospores that survive under hostile conditions. The 3D 'cell-in-shell' structures-coined as 'artificial spores'-enable modulation and control over cellular metabolism, such as control of cell division, resistance to external stresses, and surface-functionalizability, providing a useful platform for applications, including cell-based sensors, cell therapy, regenerative medicine, as well as for fundamental studies on cellular metabolism at the single-cell level and cell-to-cell communications. This Concept focuses on chemical approaches to single-cell encapsulation with artificial shells for creating artificial spores, including cross-linked layer-by-layer assembly, bioinspired mineralization, and mussel-inspired polymerization. The current status and future prospects of this emerging field are also discussed.

Assembly of bacteriophage into functional materials.

For the last decade, the fabrication of ordered structures of phage has been of great interest as a means of utilizing the outstanding biochemical properties of phage in developing useful materials. Combined with other organic/inorganic substances, it has been demonstrated that phage is a superior building block for fabricating various functional devices, such as the electrode in lithium-ion batteries, photovoltaic cells, sensors, and cell-culture supports. Although previous research has expanded the utility of phage when combined with genetic engineering, most improvements in device functionality have relied upon increases in efficiency owing to the compact, more densely packable unit size of phage rather than on the unique properties of the ordered nanostructures themselves. Recently, self-templating methods, which control both thermodynamic and kinetic factors during the deposition process, have opened up new routes to exploiting the ordered structural properties of hierarchically organized phage architectures. In addition, ordered phage films have exhibited unexpected functional properties, such as structural color and optical filtering. Structural colors or optical filtering from phage films can be used for optical phage-based sensors, which combine the structural properties of phage with target-specific binding motifs on the phage-coat proteins. This self-templating method may contribute not only to practical applications, but also provide insight into the fundamental study of biomacromolecule assembly in in vivo systems under complicated and dynamic conditions.

Chemical control of yeast cell division by cross-linked shells of catechol-grafted polyelectrolyte multilayers.

The chemical control of cell division has attracted much attention in the areas of single cell-based biology and high-throughput screening platforms. A mussel-inspired cytocompatible encapsulation method for achieving a "cell-division control" with cross-linked layer-by-layer (LbL) shells is developed. Catechol-grafted polyethyleneimine and hyaluronic acid are chosen as polyelectrolytes for the LbL process, and the cross-linking of polyelectrolytes is performed at pH 8.5. Cell division is controlled by the number of the LbL nanolayers and cross-linking reaction. We also suggest a new measuring unit, t-2.0 OD 600, for quantifying "cell-division timing" based on microbial growth kinetics.

Bioinspired, cytocompatible mineralization of silica-titania composites: thermoprotective nanoshell formation for individual chlorella cells.

Hard-shell case: Using a (RKK)4 D8 peptide allows mineralization to occur under cytocompatible conditions. Thus individual Chlorella cells could be encapsulated within a SiO2 -TiO2 nanoshell with high cell viability (87 %). The encapsulated Chlorella showed an almost threefold increase in their thermo-tolerance after 2 h at 45 °C.

Cytocompatible encapsulation of individual Chlorella cells within titanium dioxide shells by a designed catalytic peptide.

The individual encapsulation of living cells has a great impact on the areas of single cell-based sensors and devices as well as fundamental studies in single cell-based biology. In this work, living Chlorella cells were encapsulated individually with abiological, functionalizable TiO(2), by a designed catalytic peptide that was inspired by biosilicification of diatoms in nature. The bioinspired cytocompatible reaction conditions allowed the encapsulated Chlorella cells to maintain their viability and original shapes. After formation of the TiO(2) shells, the shells were postfunctionalized by using catechol chemistry. Our work suggests a bioinspired approach to the interfacing of individual living cells with abiological materials in a controlled manner.

Diffusion-Controlled Crystallization of Calcium Phosphate in a Hydrogel toward a Homogeneous Octacalcium Phosphate/Agarose Composite.

Diffusion-controlled crystallization in a hydrogel has been investigated to synthesize organic/inorganic hybrid composites and obtain a fundamental understanding of the detailed mechanism of biomineralization. Although calcium phosphate/hydrogel composites have been intensively studied and developed for the application of bone substitutes, the synthesis of homogeneous and integrated composites remains challenging. In this work, diffusion-controlled systems were optimized by manipulating the calcium ion flux at the interface, concentration gradient, and diffusion coefficient to synthesize homogeneous octacalcium phosphate/hydrogel composites with respect to the crystal morphology and density. The ion flux and local pH play an important role in determining the morphology, density, and phase of the crystals. This study suggests a model system that can reveal the relation between local conditions and the resulting crystal phase in diffusion-limited systems and provides a synthetic method for homogeneously organized organic/inorganic composites.

Mussel-inspired encapsulation and functionalization of individual yeast cells.

The individual encapsulation of living cells has a great impact on the area of cell-based sensors and devices as well as fundamental studies in cell biology. In this work, living yeast cells were individually encapsulated with functionalizable, artificial polydopamine shells, inspired by an adhesive protein in mussels. Yeast cells maintained their viability within polydopamine, and the cell cycle was controlled by the thickness of the shells. In addition, the artificial shells aided the cell in offering much stronger resistance against foreign aggression, such as lyticase. After formation of the polydopamine shells, the shells were functionalized with streptavidin by utilizing the chemical reactivity of polydopamine, and the functionalized cells were biospecifically immobilized onto the defined surfaces. Our work suggests a biomimetic approach to the encapsulation and functionalization of individual living cells with covalently bonded, artificial shells.

Glaucoma Progression and its Relationship with Corrected and Uncorrected Intraocular Pressure in Eyes with History of Refractive Corneal Surgery.

PURPOSE: To investigate glaucoma progression and its relationship with corrected and uncorrected intraocular pressure (IOP) in patients with a history of refractive corneal surgery (RCS). METHODS: Totally, 56 eyes of 56 primary open-angle glaucoma patients who had a history of RCS were included. Mean keratometry and central corneal thickness were measured at the time of glaucoma diagnosis. Three IOP measurements, i.e., uncorrected IOP (UCIOP) and corrected IOP calculated by applying the Kohlhaas (CKIOP) and Ehlers (CEIOP) formulas, were used. Participants were categorized into two groups (low-teen [<15 mmHg] and high-teen [≥15 mmHg] groups) according to the mean follow-up IOP, in each of the three IOP measurements. Glaucoma progression was determined based on either structurally or functionally by stereoscopic optic disc/retinal nerve fiber layer photographs or visual field tests. Kaplan-Meier survival curves were compared between the low-teens group and high-teens group for each IOP measurement. Risk factors for glaucoma progression, including uncorrected and corrected IOP measurements, were analyzed using a Cox proportional-hazards model. RESULTS: Among total eyes, glaucoma progression was found in 19 (34%) eyes during 4.3 years of mean follow-up period. Individual values of uncorrected IOP and corrected IOP showed significant differences (p < 0.001). Incidence of glaucoma progression was higher in high-teens group than in low-teens group by corrected IOP values (both Kohlhaas and Ehlers, p = 0.006, 0.003), but not by uncorrected IOP values (p = 0.749). The cumulative probability of glaucoma progression was significantly greater in the high-teens group than in the low-teens group using the Elhers formula determined by Kaplan-Meier analysis (p = 0.030). CONCLUSIONS: For glaucoma eyes with history of RCS, high-teen group by corrected mean follow-up IOP showed more glaucoma progression than low-teen group. More intensive IOP-lowering treatment will be required for patient with high-teen follow-up corrected IOP to prevent glaucoma progression.

Peptide-catalyzed, bioinspired silicification for single-cell encapsulation in the imidazole-buffered system.

Inspired by biosilicification of glass sponges, we designed a catalytic peptide, which formed silica structures in the imidazole-buffered solution. The peptide was adsorbed selectively onto the surface of yeast cells, and the bioinspired silicification led to the formation of a cytoprotective silica shell on individual yeast cells.

Micro/Nanostructured Films and Adhesives for Biomedical Applications.

The advanced technologies available for micro/nanofabrication have opened new avenues for interdisciplinary approaches to solve the unmet medical needs of regenerative medicine and biomedical devices. This review highlights the recent developments in micro/nanostructured adhesives and films for biomedical applications, including waterproof seals for wounds or surgery sites, drug delivery, sensing human body signals, and optical imaging of human tissues. We describe in detail the fabrication processes required to prepare the adhesives and films, such as tape-based adhesives, nanofilms, and flexible and stretchable film-based electronic devices. We also discuss their biomedical functions, performance in vitro and in vivo, and the future research needed to improve the current systems.

Chemical sporulation and germination: cytoprotective nanocoating of individual mammalian cells with a degradable tannic acid-FeIII complex.

Individual mammalian cells were coated with cytoprotective and degradable films by cytocompatible processes maintaining the cell viability. Three types of mammalian cells (HeLa, NIH 3T3, and Jurkat cells) were coated with a metal-organic complex of tannic acid (TA) and ferric ion, and the TA-Fe(III) nanocoat effectively protected the coated mammalian cells against UV-C irradiation and a toxic compound. More importantly, the cell proliferation was controlled by programmed formation and degradation of the TA-Fe(III) nanocoat, mimicking the sporulation and germination processes found in nature.

Layer-by-layer-based silica encapsulation of individual yeast with thickness control.

In the area of cell-surface engineering with nanomaterials, the metabolic and functional activities of the encapsulated cells are manipulated and controlled by various parameters of the artificial shells that encase the cells, such as stiffness and elasticity, thickness, and porosity. The mechanical durability and physicochemical stability of inorganic shells prove superior to layer-by-layer-based organic shells with regard to cytoprotection, but it has been difficult to vary the parameters of inorganic shells including their thickness. In this work, we combine the layer-by-layer technique with a process of bioinspired silicification to control the thickness of the silica shells that encapsulate yeast Saccharomyces cerevisiae cells individually, and investigate the thickness-dependent microbial growth.

Organic/inorganic double-layered shells for multiple cytoprotection of individual living cells.

The cytoprotection of individual living cells under in vitro and daily-life conditions is a prerequisite for various cell-based applications including cell therapy, cell-based sensors, regenerative medicine, and even the food industry. In this work, we use a cytocompatible two-step process to encapsulate Saccharomyces cerevisiae in a highly uniform nanometric (<100 nm) shell composed of organic poly(norepinephrine) and inorganic silica layers. The resulting cell-in-shell structure acquires multiple resistance against lytic enzyme, desiccation, and UV-C irradiation. In addition to the UV-C filtering effect of the double-layered shell, the biochemical responses of the encapsulated yeast are suggested to contribute to the observed UV-C tolerance. This work offers a chemical tool for cytoprotecting individual living cells under multiple stresses and also for studying biochemical behavior at the cellular level.