Friction-reduction effect of the hierarchical surface microstructure of carrion beetle by controlling the real contact area.

The discovery and elucidation of the surface microstructure functions of living organisms are crucial to resolving issues, such as friction. We newly discovered that Necrophila japonica, a type of carrion beetle that lives on the ground surface, exhibited a hierarchical surface microstructure comprising a submicron-sized wrinkle structure on top of a micron-sized microstructure. The surface microstructure of this beetle improved wettability but did not exhibit superhydrophobicity, a well-known function of hierarchical structures, so it was expected to have a different function. By combining the insights in the field of structural mechanics that avoidance of stress concentration by the structural geometry affects deformation with the basic principles of friction, the frictional properties and mechanisms of the hierarchical surface microstructure of carrion beetle were investigated. The measurements of frictional force indicated that the mimicked structure exerted lower frictional forces than flat and single-layer microstructure surfaces. Analysis of finite element method simulations showed that even though the mimicked structure was prone to pressure concentration due to small contact points, the surface contact pressure was reduced more than that of the single-layer structure by hierarchical load dispersion like that of metamaterials. As a result, the suppression of the increase in the real contact area due to deformation suppression contributed to effective friction-reduction. The effective friction-reduction by the hierarchical structure provides not only new insight into the surface microstructure function of various organisms, but also new design for the lubricant-free friction reduction that has been the focus of attention in carbon neutrality and other fields.

A New Concept for an Adhesive Material Inspired by Clingfish Sucker Nanofilaments.

Underwater adhesive materials are in high demand in various fields, and fish species with sucker disks have attracted attention due to their superior performance and interesting structures. The clingfish, in particular, is widely known for using hierarchical sucker disk structures to demonstrate rapid and strong adhesion to rocky surfaces under strong currents. We examined the combination of nanofilaments and mucus in the clingfish sucker disk. Nanofilaments reinforce mucus adhesion force by reducing the compliance without affecting the contact area. We prepared structures from hard polymers and soft polydimethylsiloxane (PDMS) that mimicked clingfish sucker nanofilaments and mucus, with these biomimetic structures showing significant adhesion force underwater. Furthermore, the hardness and length of the nanofilaments and Young's modulus and thickness of the mucus-mimicking PDMS layer had critical effects on the adhesion force. According to the results, clingfish nanofilaments act as hard bracing for the soft mucus, and the structural combination of the conflicting characteristics of hardness and softness, thus achieved, is crucial for strong adhesion.

Microscopy and biomimetics: the NanoSuit® method and image retrieval platform.

This review aims to clarify a suitable method towards achieving next-generation sustainability. As represented by the term 'Anthropocene', the Earth, including humans, is entering a critical era; therefore, science has a great responsibility to solve it. Biomimetics, the emulation of the models, systems and elements of nature, especially biological science, is a powerful tool to approach sustainability problems. Microscopy has made great progress with the technology of observing biological and artificial materials and its techniques have been continuously improved, most recently through the NanoSuit® method. As one of the most important tools across many facets of research and development, microscopy has produced a large amount of accumulated digital data. However, it is difficult to extract useful data for making things as biomimetic ideas despite a large amount of biological data. Here, we would like to find a way to organically connect the indispensable microscopic data with the new biomimetics to solve complex human problems.

Hydrophobic-hydrophilic crown-like structure enables aquatic insects to reside effectively beneath the water surface.

Various insects utilise hydrophobic biological surfaces to live on the surface of water, while other organisms possess hydrophilic properties that enable them to live within a water column. Dixidae larvae reside, without being submerged, just below the water surface. However, little is known about how these larvae live in such an ecological niche. Herein, we use larvae of Dixa longistyla (Diptera: Dixidae) as experimental specimens and reveal their characteristics. A complex crown-like structure on the abdomen consists of hydrophobic and hydrophilic elements. The combination of these contrasting features enables the larvae to maintain their position as well as to move unidirectionally. Their hydrophobic region leverages water surface tension to function as an adhesive disc. By using the resistance of water, the hydrophilic region serves as a rudder during locomotion.

Effect of the Microstructures on Vulcanized Rubber Frictions.

Vulcanized rubber is widely used in a wide range of applications because of its flexibility, durability, sealing properties, and high degree of friction. However, this high degree of friction can also become an issue, as it leads to the wearing and breakage of parts. In this report, we investigated the effects of the vulcanized rubber microstructures on friction force by using simple, anisotropic microstructures. The line and space master microstructures were prepared from a photoresist, and the structures were transferred to PDMS, PSt, and then Ni. After surface modification of the Ni microstructures by TEOS, the vulcanized rubber microstructures were fabricated by a simple hot press process with the TEOS-coated Ni microstructure molds. The structural parameters of the vulcanized rubber line and space microstructures were found to be successfully varied by elongation, and the structural deformations were also investigated by FEM simulations. Measurements of the frictional force of the vulcanized rubber microstructures revealed the friction coefficient was reduced by the surface microstructures and was affected by the directions of the contact because of the microstructure anisotropy. The reason for of these results can be explained by the changes in the contact area and hysteresis friction. These results suggest that the friction coefficients of vulcanized rubbers can be reduced by the simple surface microstructures that are applicable to a wide range of fields.

In situ elemental analyses of living biological specimens using 'NanoSuit' and EDS methods in FE-SEM.

Energy dispersive X-ray spectroscopy (EDS) carried out alongside scanning electron microscopy (SEM) is a common technique for elemental analysis. To investigate "wet" biological specimens, complex pre-treatments are required to stabilize them under the high vacuum conditions of high-resolution SEM. These often produce unwanted artifacts. We have previously reported that the polymerization of natural surface substances on organisms by the electron beam of the SEM setup or by plasma irradiation causes a nano-scale layer to form-called a "NanoSuit"-that can act as a barrier and keep organisms alive and hydrated in a field-emission SEM system. In the study reported herein, we examined the suitability of the NanoSuit method for elemental analyses of biological specimens by EDS. We compared experimental results for living Drosophila larvae and Aloe arborescens specimens prepared by the NanoSuit method and by conventional fixation. The NanoSuit method allowed accurate detection of the elemental compositions at high resolution. By contrast, specimens prepared by the conventional fixation method displayed additional EDS signals corresponding to the elements in the chemicals involved in the fixation process. Our results demonstrate that the NanoSuit method is useful for studying hydrous samples via EDS and SEM, particularly in biological sciences.

Imaging dataset of fresh hydrous plants obtained by field-emission scanning electron microscopy conducted using a protective NanoSuit.

Although scanning electron microscopy (SEM) can generate high-resolution images of nanosized objects, it requires a high vacuum to do so, which precludes direct observations of living organisms and often produces unwanted structural changes. It has previously been reported that a simple surface modification gives rise to a nanoscale layer, termed the "NanoSuit", which can keep small animals alive under the high vacuum required for field-emission scanning electron microscopy (FE-SEM). We have previously applied this technique to plants, and successfully observed healthy petals in a fully hydrated state using SEM. The flower petals protected with the NanoSuit appeared intact, although we still lack a fundamental understanding of the images of other plants observed using FE-SEM. This report presents and evaluates a rich set of images, acquired using the NanoSuit, for a taxonomically diverse set of plant species. This dataset of images allows the surface features of various plants to be analyzed and thus provides a further complementary morphological profile. Image data can be accessed and viewed through Figshare (https://doi.org/10.6084/m9.figshare.c.4446026.v1).

The Friction Properties of Firebrat Scales.

Friction is an important subject for sustainability due to problems that are associated with energy loss. In recent years, micro- and nanostructured surfaces have attracted much attention to reduce friction; however, suitable structures are still under consideration. Many functional surfaces are present in nature, such as the friction reduction surfaces of snake skins. In this study, we focused on firebrats, Thermobia domestica, which temporary live in narrow spaces, such as piled papers, so their body surface (integument) is frequently in contact with surrounding substrates. We speculate that, in addition to optical, cleaning effects, protection against desiccation and enemies, their body surface may be also adapted to reduce friction. To investigate the functional effects of the firebrat scales, firebrat surfaces were observed using a field-emission scanning electron microscope (FE-SEM) and a colloidal probe atomic force microscope (AFM). Results of surface observations by FE-SEM revealed that adult firebrats are entirely covered with scales, whose surfaces have microgroove structures. Scale groove wavelengths around the firebrat's head are almost uniform within a scale but they vary between scales. At the level of single scales, AFM friction force measurements revealed that the firebrat scale reduces friction by decreasing the contact area between scales and a colloidal probe. The heterogeneity of the scales' groove wavelengths suggests that it is difficult to fix the whole body on critical rough surfaces and may result in a "fail-safe" mechanism.

A 'NanoSuit' successfully protects petals of cherry blossoms in high vacuum: examination of living plants in an FE-SEM.

Land plants have evolved on dry land and developed surface barriers to protect themselves from environmental stresses. We have previously reported that polymerization of a natural extracellular substance (ECS) on the outer surface of animals by electron beam or plasma irradiation, can give rise to a nano-scale layer, termed the "NanoSuit", which can keep small animals alive under the high vacuum of a scanning electron microscope (SEM). In the present research, we have focused on plants, using petals of cherry blossoms, as experimental specimens and examined their behavior under high vacuum conditions. Experiments on healthy living petals have demonstrated that without any pre-treatment, the overall morphology of specimens is well preserved and intact after imaging in an SEM, suggesting that natural substances on the petal surface behave like animal ECS and form a NanoSuit following irradiation with an electron beam. Furthermore, we have shown that the surface material can be extracted with chloroform and polymerized into a free-standing membrane by plasma irradiation. From our results, we conclude that surface materials, which have the ability to prevent water loss under natural conditions, increase the barrier ability and can protect plants under high vacuum conditions.

A modified 'NanoSuit®' preserves wet samples in high vacuum: direct observations on cells and tissues in field-emission scanning electron microscopy.

Although field-emission scanning electron microscopy (FE-SEM) has proven very useful in biomedical research, the high vacuum required (10-3 to 10-7 Pa) precludes direct observations of living cells and tissues at high resolution and often produces unwanted structural changes. We have previously described a method that allows the investigator to keep a variety of insect larvae alive in the high vacuum environment of the electron microscope by encasing the organisms in a thin, vacuum-proof suit, the 'NanoSuit®'. However, it was impossible to protect wet tissues freshly excised from intact organisms or cultured cells. Here we describe an improved 'NanoSuit' technique to overcome this limitation. We protected the specimens with a surface shield enhancer (SSE) solution that consists of glycerine and electrolytes and found that the fine structure of the SSE-treated specimens is superior to that of conventionally prepared specimens. The SSE-based NanoSuit affords a much stronger barrier to gas and/or liquid loss than the previous NanoSuit did and, since it allows more detailed images, it could significantly help to elucidate the 'real' organization of cells and their functions.

Uphill Water Transport on a Wettability-Patterned Surface: Experimental and Theoretical Results.

In nature, there exist many functional water-controlling surfaces, such as the water-repellent surface of lotus leaves, the superhydrophobic water-adhesive surface of rose petals, the water-harvesting surface of a beetle's back, and the water-transporting surface of the legs of Ligia exotica. These natural surfaces suggest that surface chemistry and hierarchical structures are essential for controlling the water behavior. We have reported the preparation of superhydrophobic and antireflection silicon nanospike-array structures using self-organized honeycomb-patterned films as three-dimensional dry-etching masks. Moreover, the surface wettability of the silicon nanospike-array structures can be easily transformed from superhydrophobic to superhydrophilic by changes in the surface chemistry. In this report, we show the preparation of water-controlling surfaces, such as water-harvesting and water-transporting surfaces, by the wettability patterning of silicon nanostructured surfaces. We prepared honeycomb-patterned films for dry-etching masks made from polystyrene and an amphiphilic polymer by casting a chloroform solution. After the fixation of the top layer of the honeycomb-patterned films on a single-crystal silicon substrate, reactive ion etching was performed. The as-prepared silicon nanospike-array structure showed superhydrophobicity, and the water contact angles were over 170°. After UV-O3 treatment with photomasks, only the UV-irradiated surfaces showed superhydrophilicity, suggesting that we can obtain superhydrophobic- and superhydrophilic-patterned surfaces for which the patterns are the same as those of the photomasks. On the basis of these wettability-patterned surfaces, we demonstrated water harvesting by superhydrophilic dot-patterned surfaces and water transportation against gravity by superhydrophilic triangular-patterned surfaces. In particular, we investigated uphill water transport through the motion of droplets on tilting slopes based on the equation of motion. These results suggested that we can obtain superior microfluidic devices suitable for various applications through the use of optional wettability patterns.

Paclitaxel-Releasing Thin Biodegradable Film for Prevention of Bleb Avascularity Without Compromising Filtration in Rabbits.

PURPOSE: A honeycomb-patterned film (HPF) prevents bleb scarring and mitomycin C (MMC)-related bleb avascularity in a rabbit model of filtration surgery. In this study, we examined whether a HPF-releasing paclitaxel (PTX) can prevent bleb avascularity without compromising filtration. METHODS: Filtration surgery was performed in one eye of rabbits. A 14-µm thick HPF made from poly(L-lactide-co-ε-caprolactone) was placed subconjunctivally over the filtration site with the honeycomb surface turned toward the subconjunctival Tenon tissue. The rabbits were divided into four groups (n = 5 each): 1, HPF with no drug; 2, HPF + PTX 50 µg; 3, HPF + 5 µg; 4, HPF + 0.5 µg. Intraocular pressure (IOP) measurements and bleb evaluations using ultrasound biomicroscopy were performed periodically for 4 weeks followed by histological examination. A longer follow-up study (12 weeks) was performed for group 4 (experiment 2; n = 8). RESULTS: Among all groups at the 4-week follow up, two blebs failed in group 1. The postoperative IOP decrease was significantly greater in PTX-treated eyes than in group 1. The bleb avascular area persisted for 4 weeks in groups 2 and 3. However, no avascular area was observed in groups 1 and 4 at 4 weeks postoperatively. Histology showed minimal fibrosis at the filtration site in all the PTX groups. In experiment 2, some blebs became flatter starting at 10 weeks after surgery. CONCLUSIONS: PTX released from HPF promoted bleb survival and IOP decrease. The lowest dose of PTX (0.5 µg) was effective at preventing bleb avascularity without compromising filtration.

Dehydrogenative polymerization of coniferyl alcohol in artificial polysaccharides matrices: effects of xylan on the polymerization.

To elucidate the influence of wood polysaccharide components on lignin formation in vitro, models for polysaccharide matrix in wood secondary cell wall were fabricated from two types of bacterial cellulosic films, flat film (FBC) and honeycomb-patterned film (HPBC), as basic frameworks by depositing xylan onto the films. An endwise type of dehydrogenative polymerization, "Zutropfverfahren", of coniferyl alcohol was attempted in the films with/without xylan. The resultant dehydrogenation polymer (DHP) was generated inside and outside xylan-deposited films, whereas DHP was deposited only outside the films without xylan. The amount of the generated DHP in the xylan-deposited films was larger than that in the films without xylan. The frequency of 8-O-4' interunitary linkage in DHP was also increased by the xylan deposition. These results suggest that xylan acts as a scaffold for DHP deposition in polysaccharides matrix and as a structure regulator for the formation of the 8-O-4' linkage. In addition, mechanical properties, i.e., tensile strength and modulus of elasticity (MOE), of both cellulosic films were found to be augmented by the deposition of xylan and DHP. Especially, DHP deposition remarkably enhanced MOE. Such effects of xylan on DHP formation and augmentation of mechanical strength were clearly observed for HPBC, revealing that HPBC is a promising framework model to investigate wood cell wall formation in vitro.

A 'NanoSuit' surface shield successfully protects organisms in high vacuum: observations on living organisms in an FE-SEM.

Although extremely useful for a wide range of investigations, the field emission scanning electron microscope (FE-SEM) has not allowed researchers to observe living organisms. However, we have recently reported that a simple surface modification consisting of a thin extra layer, termed 'NanoSuit', can keep organisms alive in the high vacuum (10(-5) to 10(-7) Pa) of the SEM. This paper further explores the protective properties of the NanoSuit surface-shield. We found that a NanoSuit formed with the optimum concentration of Tween 20 faithfully preserves the integrity of an organism's surface without interfering with SEM imaging. We also found that electrostatic charging was absent as long as the organisms were alive, even if they had not been coated with electrically conducting materials. This result suggests that living organisms possess their own electrical conductors and/or rely on certain properties of the surface to inhibit charging. The NanoSuit seems to prolong the charge-free condition and increase survival time under vacuum. These findings should encourage the development of more sophisticated observation methods for studying living organisms in an FE-SEM.

Breath Figures of Nanoscale Bricks: A Universal Method for Creating Hierarchic Porous Materials from Inorganic Nanoparticles Stabilized with Mussel-Inspired Copolymers.

High-performance catalysts and photovoltaics are required for building an environmentally sustainable society. Because catalytic and photovoltaic reactions occur at the interfaces between reactants and surfaces, the chemical, physical, and structural properties of interfaces have been the focus of much research. To improve the performance of these materials further, inorganic porous materials with hierarchic porous architectures have been fabricated. The breath figure technique allows preparing porous films by using water droplets as templates. In this study, a valuable preparation method for hierarchic porous inorganic materials is shown. Hierarchic porous materials are prepared from surface-coated inorganic nanoparticles with amphiphilic copolymers having catechol moieties followed by sintering. Micron-scale pores are prepared by using water droplets as templates, and nanoscale pores are formed between the nanoparticles. The fabrication method allows the preparation of hierarchic porous films from inorganic nanoparticles of various shapes and materials.

Capillary rise on legs of a small animal and on artificially textured surfaces mimicking them.

The wharf roach Ligia exotica is a small animal that lives by the sea and absorbs water from the sea through its legs by virtue of a remarkable array of small blades of micron scale. We find that the imbibition dynamics on the legs is rather complex on a microscopic scale, but on a macroscopic scale the imbibition length seems to simply scale linearly with elapsed time. This unusual dynamics of imbibition, which usually slows down with time, is advantageous for long-distance water transport and results from repetition of unit dynamics. Inspired by the remarkable features, we study artificially textured surfaces mimicking the structure on the legs of the animal. Unlike the case of the wharf roach, the linear dynamics were not reproduced on the artificial surfaces, which may result from more subtle features on the real legs that are not faithfully reflected on the artificial surfaces. Instead, the nonlinear dynamics revealed that hybrid structures on the artificial surfaces speed up the water transport compared with non-hybrid ones. In addition, the dynamics on the artificial surfaces turn out to be well described by a composite theory developed here, with the theory giving useful guiding principles for designing hybrid textured surfaces for rapid imbibition and elucidating physical advantages of the microscopic design on the legs.

Dressing living organisms in a thin polymer membrane, the NanoSuit, for high-vacuum FE-SEM observation.

Scanning electron microscopy (SEM) has made remarkable progress and has become an essential tool for observing biological materials at microscopic level. However, various complex procedures have precluded observation of living organisms to date. Here, a new method is presented by which living organisms can be observed by field emission (FE)-SEM. Using this method, active movements of living animals were observed in vacuo (10(-5)-10(-7) Pa) by protecting them with a coating of thin polymer membrane, a NanoSuit, and it was found that the surface fine structure of living organisms is very different from that of traditionally fixed samples. After observation of mosquito larvae in the high vacuum of the FE-SEM, it was possible to rear them subsequently in normal culture conditions. This method will be useful for numerous applications, particularly for electron microscopic observations in the life sciences.

The morphology and functions of articular chondrocytes on a honeycomb-patterned surface.

The present study investigated the potential of a novel micropatterned substrate for neocartilage formation. Articular chondrocytes were cultured on poly( ɛ-caprolactone) materials whose surfaces were either flat or honeycomb-patterned. The latter was prepared using a novel self-organization technique, while the former, was prepared by spin-coating. The chondrocytes attached and proliferated on both surfaces. On the honeycomb films, chondrocytes were found at the top surface and encased within the 10 µm pores. Meanwhile, chondrocytes on the spin-coated surface flattened out. Accumulation of DNA and keratin sulphate was comparatively higher on the honeycomb films within the first 7 days. At their respective peaks, DNA concentration increased on the honeycomb and flat surfaces by approximately 210% and 400% of their day 1 values, respectively. However, cultures on the flat surface took longer to peak. Extracellular Matrix (ECM) concentrations peaked at 900% and 320% increases for the honeycomb and flat cultures. Type II collagen was upregulated on the honeycomb and flat surfaces by as much as 28% and 25% of their day 1 values, while aggrecan was downregulated with time, by 3.4% and 7.4%. These initial results demonstrate the potential usefulness of honeycomb-based scaffolds during early cultures neocartilage and soft tissue engineering.

Micropatterned culture of HepG2 spheroids using microwell chip with honeycomb-patterned polymer film.

Microwell chip culture is a promising technique for the generation of homogenous spheroids. We investigated the relationship between the structure of the bottom surface of microwell chip and the properties of HepG2 spheroid. We developed a microwell chip, the bottom surface of which consisted of a honeycomb-patterned polymer film (honeycomb film) that had a regular porous structure (HF chip). The chip comprised 270 circular microwells; each microwell was 600 µm in diameter and 600 µm in depth. At the center of the honeycomb film, an area, 200 µm in diameter, was modified with collagen to facilitate cell adhesion. With the exception of the collagen-coated area, the entire microwell was modified with polyethylene glycol to eliminate cell adhesion. HepG2 cells formed uniform spheroids when cultured in the microwells of HF chip. Furthermore, the cells passed through the porous structure of honeycomb film and formed spheroids at its opposite side. The spheroid growth of HepG2 cells cultured in HF chip was greater than that when the cells were culture in a microwell chip, the bottom surface of which was made of poly-methylmethacrylate (PMMA chip). The albumin secretion activity of HepG2 spheroids in HF chip was equal to that in PMMA chip. These results indicate that the microwell bottom with a porous structure enhances the cell growth and maintains well the spheroid function. Thus, HF chip is a promising platform for spheroid cell culture.

Fabrication of gold nanoparticle-polymer composite particles with raspberry, core-shell and amorphous morphologies at room temperature via electrostatic interactions and diffusion.

Composite particles with varying morphologies composed of gold nanoparticles (Au NPs) and polymers were fabricated based on a combination of electrostatic interactions between the polymer particles and Au NPs and diffusion processes. The positively charged polymer particles were prepared from amino-terminated polystyrene (PS-NH2) and amino-terminated 1,2-polybutadiene (PB-NH2). Adsorption of citrate-stabilized Au NPs resulted in three different distribution states of Au NPs in the polymer particles, depending on the glass transition temperature (Tg) and molecular weight of the polymer. The adsorption of Au NPs onto PS-NH2 particles produced raspberry-like composite particle morphologies, while the NPs instead diffused into the PB-NH2 particles, since the Tg of PB-NH2 is below room temperature. The diffusion of Au NPs could be controlled by varying the molecular weight of the PB-NH2 and the diameter of the NPs, and both core-shell and amorphous distributions were successfully achieved.