Sequenced Somatic Cell Reprogramming and Differentiation Inside Nested Hydrogel Droplets.

The efficient genesis of pluripotent cells or therapeutic cells for regenerative medicine involves several external manipulations and conditioning protocols, which drives down clinical applicability. Automated programming of the genesis by microscale physical forces and chronological biochemistry can increase clinical success. The design and fabrication of nested polysaccharide droplets (millimeter-sized) with cell sustaining properties of natural tissues and intrinsic properties for time and space evolution of cell transformation signals between somatic cells, pluripotent cells and differentiated therapeutic cells in a swift and efficient manner without the need for laborious external manipulation are reported. Cells transform between phenotypic states by having single and double nested droplets constituted with extracellular matrix proteins and reprogramming, and differentiation factors infused chronologically across the droplet space. The cell transformation into germ layer cells and bone cells is successfully tested in vitro and in vivo and promotes the formation of new bone tissues. Thus, nested droplets with BMP-2 loaded guests synthesize mineralized bone tissue plates along the length of a cranial non-union bone defect at 4 weeks. The advantages of sequenced somatic cell reprogramming and differentiation inside an individual hydrogel module without external manipulation, promoted by formulating tissue mimetic physical, mechanical, and chemical microenvironments are shown.

Natural History Collections as Inspiration for Technology.

Living organisms are the ultimate survivalists, having evolved phenotypes with unprecedented adaptability, ingenuity, resourcefulness, and versatility compared to human technology. To harness these properties, functional descriptions and design principles from all sources of biodiversity information must be collated - including the hundreds of thousands of possible survival features manifest in natural history museum collections, which represent 12% of total global biodiversity. This requires a consortium of expert biologists from a range of disciplines to convert the observations, data, and hypotheses into the language of engineering. We hope to unite multidisciplinary biologists and natural history museum scientists to maximize the coverage of observations, descriptions, and hypotheses relating to adaptation and function across biodiversity, to make it technologically useful. This is to be achieved by developments in meta- taxonomic classification, phylogenetics, systematics, biological materials research, structure and morphological characterizations, and ecological data gathering from the collections - the aim being to identify and catalogue features essential for good biomimetic design.

Differences in Nanostructure and Hydrophobicity of Cicada (Cryptotympana atrata) Forewing Surface with the Distribution of Precipitation.

Although the cicada wing has a variety of functions and the nanostructure and surface properties of many species have been extensively investigated, there are no reports investigating diversity of nanostructures and wetting properties within a single species collected at locations with different rainfall conditions. In this study, the hydrophobicity and nanostructure dimensions of the forewing surface of Cryptotympana atrata were measured, based on specimens collected from 12 distributions with varying precipitation averages in China and Japan. The relationships among hydrophobicity, nanostructures, and precipitation were analyzed, and the adaption of hydrophobic nanostructures under different wet environments is discussed. The precipitation of locations in the years the samples of C. atrata were collected only has an effect on the diameter and spacing of wing surface nanostructure, and the multiple years of precipitation may have an influence on the basic diameter and spacing, as well as the height of protrusions. The rougher the wing surface, the stronger the hydrophobicity which was observed from samples taken where the rainfall conditions of the collection years are high. To our knowledge, this is one special example providing evidence of hydrophobic nanostructures found on a biological surface of a single species which shows adaption for specific wet environments.

Insect Analogue to the Lotus Leaf: A Planthopper Wing Membrane Incorporating a Low-Adhesion, Nonwetting, Superhydrophobic, Bactericidal, and Biocompatible Surface.

Nature has produced many intriguing and spectacular surfaces at the micro- and nanoscales. These small surface decorations act for a singular or, in most cases, a range of functions. The minute landscape found on the lotus leaf is one such example, displaying antiwetting behavior and low adhesion with foreign particulate matter. Indeed the lotus leaf has often been considered the "benchmark" for such properties. One could expect that there are animal counterparts of this self-drying and self-cleaning surface system. In this study, we show that the planthopper insect wing (Desudaba danae) exhibits a remarkable architectural similarity to the lotus leaf surface. Not only does the wing demonstrate a topographical likeness, but some surface properties are also expressed, such as nonwetting behavior and low adhering forces with contaminants. In addition, the insect-wing cuticle exhibits an antibacterial property in which Gram-negative bacteria (Porphyromonas gingivalis) are killed over many consecutive waves of attacks over 7 days. In contrast, eukaryote cell associations, upon contact with the insect membrane, lead to a formation of integrated cell sheets (e.g., among human stem cells (SHED-MSC) and human dermal fibroblasts (HDF)). The multifunctional features of the insect membrane provide a potential natural template for man-made applications in which specific control of liquid, solid, and biological contacts is desired and required. Moreover, the planthopper wing cuticle provides a "new" natural surface with which numerous interfacial properties can be explored for a range of comparative studies with both natural and man-made materials.

Diversity of Cuticular Micro- and Nanostructures on Insects: Properties, Functions, and Potential Applications.

Insects exhibit a fascinating and diverse range of micro- and nanoarchitectures on their cuticle. Beyond the spectacular beauty of such minute structures lie surfaces evolutionarily modified to act as multifunctional interfaces that must contend with a hostile, challenging environment, driving adaption so that these can then become favorable. Numerous cuticular structures have been discovered this century; and of equal importance are the properties, functions, and potential applications that have been a key focus in many recent studies. The vast range of insect structuring, from the most simplistic topographies to the most elegant and geometrically complex forms, affords us with an exhaustive library of natural templates and free technologies to borrow, replicate, and employ for a range of applications. Of particular importance are structures that imbue cuticle with antiwetting properties, self-cleaning abilities, antireflection, enhanced color, adhesion, and antimicrobial and specific cell-attachment properties.

Use of Tethered Hydrogel Microcoatings for Mesenchymal Stem Cell Equilibrium, Differentiation, and Self-Organization into Microtissues.

Therapeutic adult mesenchymal stem cells (MSCs) lose multipotency and multilineage specialization in culture and after transplantation due to the absence of complex biological architecture. Here, it is shown that a transient ultrathin covering of permeable biomaterial can be differentially formulated to either preserve multipotency or induce multidifferentiation. Accordingly, populations of single, spherical MSCs in suspended media with high selectivity and specificity can be coated. Assembly of single, double, and triple hydrogel layers at MSC membranes is initiated by first attaching MSC-specific immunoglobulins onto CD90 or Stro-1 receptors and UEA-1 and soybean lectins. A secondary biotinylated immunoglobulin is targeted for avidin binding, which becomes an attractor for biotinylated alginate or hyaluronate, which are subsequently stiffened and gelled, in situ around the entire cell surface. Alginate microcoatings permeated with mobile BMP-2-induced osteospecialized tissue, vascular endothelial growth factor (VEGF) induced microcapillary formation, while microcoatings, with selected basement membrane proteins, preserve the multipotent phenotype of MSCs, for continuing rounds of culture and directed specialization. Furthermore, forced packing of microcoated MSC populations creates prototypical tissue compartments: the coating partially simulating the extracellular matrix structures. Remarkably, microcoated MSC clusters show a tremendous simulation of a common embryological tissue transformation into the epithelium. Thus, confinement of free morphology exerts another control on tissue specialization and formation.

Biomimetics for early stage biofouling prevention: templates from insect cuticles.

A biomimetic antifouling material study was carried out utilising superhydrophobic cicada and dragonfly wings replicated with a polymer (epoxy resin). They were tested in a marine biofouling study for up to 1 week in addition to biofouling assays of protein, carbohydrate and DNA absorption. The materials were compared against a commercial antifouling paint and a polymeric smooth surface constituting a control sample. The replicated surfaces demonstrated superior antifouling properties in comparison to the control and similar efficiency in DNA (10% reduction), protein and carbohydrate adsorption (15%) to the commercial anti-fouling paint. As the fabricated surfaces have roughness at the nanometre scale it is probable that the low adsorption properties, at least in the early stages, may be related to air trapped at the surface. Interestingly the most disordered replicated surface (dragonfly wing replicate) demonstrated the lowest values of absorption.

Contaminant adhesion (aerial/ground biofouling) on the skin of a gecko.

In this study, we have investigated the micro- and nano-structuring and contaminant adhesional forces of the outer skin layer of the ground dwelling gecko--Lucasium steindachneri. The lizard's skin displayed a high density of hairs with lengths up to 4 µm which were spherically capped with a radius of curvature typically less than 30 nm. The adhesion of artificial hydrophilic (silica) and hydrophobic (C18) spherical particles and natural pollen grains were measured by atomic force microscopy and demonstrated extremely low values comparable to those recorded on superhydrophobic insects. The lizard scales which exhibited a three-tier hierarchical architecture demonstrated higher adhesion than the trough regions between scales. The two-tier roughness of the troughs comprising folding of the skin (wrinkling) limits the number of contacting hairs with particles of the dimensions used in our study. The gecko skin architecture on both the dorsal and trough regions demonstrates an optimized topography for minimizing solid-solid and solid-liquid particle contact area, as well as facilitating a variety of particulate removal mechanisms including water-assisted processes. These contrasting skin topographies may also be optimized for other functions such as increased structural integrity, levels of wear protection and flexibility of skin for movement and growth. While single hair adhesion is low, contributions of many thousands of individual hairs (especially on the abdominal scale surface and if deformation occurs) may potentially aid in providing additional adhesional capabilities (sticking ability) for some gecko species when interacting with environmental substrates such as rocks, foliage and even man-made structuring.

Removal mechanisms of dew via self-propulsion off the gecko skin.

Condensation resulting in the formation of water films or droplets is an unavoidable process on the cuticle or skin of many organisms. This process generally occurs under humid conditions when the temperature drops below the dew point. In this study, we have investigated dew conditions on the skin of the gecko Lucasium steindachneri. When condensation occurs, we show that small dew drops, as opposed to a thin film, form on the lizard's scales. As the droplets grow in size and merge, they can undergo self-propulsion off the skin and in the process can be carried away a sufficient distance to freely engage with external forces. We show that factors such as gravity, wind and fog provide mechanisms to remove these small droplets off the gecko skin surface. The formation of small droplets and subsequent removal from the skin may aid in reducing microbial contact (e.g. bacteria, fungi) and limit conducive growth conditions under humid environments. As well as providing an inhospitable microclimate for microorganisms, the formation and removal of small droplets may also potentially aid in other areas such as reduction and cleaning of some surface contaminants consisting of single or multiple aggregates of particles.

A gecko skin micro/nano structure - A low adhesion, superhydrophobic, anti-wetting, self-cleaning, biocompatible, antibacterial surface.

Geckos, and specifically their feet, have attracted significant attention in recent times with the focus centred around their remarkable adhesional properties. Little attention however has been dedicated to the other remaining regions of the lizard body. In this paper we present preliminary investigations into a number of notable interfacial properties of the gecko skin focusing on solid and aqueous interactions. We show that the skin of the box-patterned gecko (Lucasium sp.) consists of dome shaped scales arranged in a hexagonal patterning. The scales comprise of spinules (hairs), from several hundred nanometres to several microns in length, with a sub-micron spacing and a small radius of curvature typically from 10 to 20 nm. This micro and nano structure of the skin exhibited ultralow adhesion with contaminating particles. The topography also provides a superhydrophobic, anti-wetting barrier which can self clean by the action of low velocity rolling or impacting droplets of various size ranges from microns to several millimetres. Water droplets which are sufficiently small (10-100 µm) can easily access valleys between the scales for efficient self-cleaning and due to their dimensions can self-propel off the surface enhancing their mobility and cleaning effect. In addition, we demonstrate that the gecko skin has an antibacterial action where Gram-negative bacteria (Porphyromonas gingivalis) are killed when exposed to the surface however eukaryotic cell compatibility (with human stem cells) is demonstrated. The multifunctional features of the gecko skin provide a potential natural template for man-made applications where specific control of liquid, solid and biological contacts is required.

Self-propulsion of dew drops on lotus leaves: a potential mechanism for self cleaning.

This study shows that condensation on the hierarchically structured lotus leaf can facilitate self-propulsion of water droplets off the surface. Droplets on leaves inclined at high angles can be completely removed from the surface by self-propulsion with the assistance of gravity. Due to the small size of mobile droplets, light breezes may also fully remove the propelled droplets, which are typically projected beyond the boundary layer of the leaf cuticle. Moreover the self-propelled droplets/condensate were able to remove contaminants (eg silica particles) from the leaf surface. The biological significance of this process may be associated with maintaining a healthy cuticle surface when the action of rain to clean the surface via the lotus effect is not possible (due to no precipitation). Indeed, the native lotus plants in this study were located in a region with extended time periods (several months) without rain. Thus, dew formation on the leaf may provide an alternative self-cleaning mechanism during times of drought and optimise the functional efficiency of the leaf surface as well as protecting the surface from long term exposure to pathogens such as bacteria and fungi.

Bactericidal activity of black silicon.

Black silicon is a synthetic nanomaterial that contains high aspect ratio nanoprotrusions on its surface, produced through a simple reactive-ion etching technique for use in photovoltaic applications. Surfaces with high aspect-ratio nanofeatures are also common in the natural world, for example, the wings of the dragonfly Diplacodes bipunctata. Here we show that the nanoprotrusions on the surfaces of both black silicon and D. bipunctata wings form hierarchical structures through the formation of clusters of adjacent nanoprotrusions. These structures generate a mechanical bactericidal effect, independent of chemical composition. Both surfaces are highly bactericidal against all tested Gram-negative and Gram-positive bacteria, and endospores, and exhibit estimated average killing rates of up to ~450,000 cells min(-1) cm(-2). This represents the first reported physical bactericidal activity of black silicon or indeed for any hydrophilic surface. This biomimetic analogue represents an excellent prospect for the development of a new generation of mechano-responsive, antibacterial nanomaterials.

Enhanced gametocyte formation in erythrocyte progenitor cells: a site-specific adaptation by Plasmodium falciparum.

Gametocytogenesis by Plasmodium falciparum is essential for transmission of the parasite from human to mosquito, yet developing gametocytes lack expression of surface proteins required for cytoadherence. Therefore, elimination from the circulation should occur unless they are sequestered in regions of low blood flow such as the extracellular spaces of the bone marrow. Our data indicate that gametocytogenesis is enhanced in the presence of erythroid progenitors found within the bone marrow. Furthermore, atomic force microscopy indicates that developing gametocytes undergo remarkable shifts in their erythrocyte membrane elasticity, which may allow them to be retained within the bone marrow until maturation.

Self-cleaning of superhydrophobic surfaces by self-propelled jumping condensate.

The self-cleaning function of superhydrophobic surfaces is conventionally attributed to the removal of contaminating particles by impacting or rolling water droplets, which implies the action of external forces such as gravity. Here, we demonstrate a unique self-cleaning mechanism whereby the contaminated superhydrophobic surface is exposed to condensing water vapor, and the contaminants are autonomously removed by the self-propelled jumping motion of the resulting liquid condensate, which partially covers or fully encloses the contaminating particles. The jumping motion off the superhydrophobic surface is powered by the surface energy released upon coalescence of the condensed water phase around the contaminants. The jumping-condensate mechanism is shown to spontaneously clean superhydrophobic cicada wings, where the contaminating particles cannot be removed by gravity, wing vibration, or wind flow. Our findings offer insights for the development of self-cleaning materials.

High-spatial-resolution mapping of superhydrophobic cicada wing surface chemistry using infrared microspectroscopy and infrared imaging at two synchrotron beamlines.

The wings of some insects, such as cicadae, have been reported to possess a number of interesting and unusual qualities such as superhydrophobicity, anisotropic wetting and antibacterial properties. Here, the chemical composition of the wings of the Clanger cicada (Psaltoda claripennis) were characterized using infrared (IR) microspectroscopy. In addition, the data generated from two separate synchrotron IR facilities, the Australian Synchrotron Infrared Microspectroscopy beamline (AS-IRM) and the Synchrotron Radiation Center (SRC), University of Wisconsin-Madison, IRENI beamline, were analysed and compared. Characteristic peaks in the IR spectra of the wings were assigned primarily to aliphatic hydrocarbon and amide functionalities, which were considered to be an indication of the presence of waxy and proteinaceous components, respectively, in good agreement with the literature. Chemical distribution maps showed that, while the protein component was homogeneously distributed, a significant degree of heterogeneity was observed in the distribution of the waxy component, which may contribute to the self-cleaning and aerodynamic properties of the cicada wing. When comparing the data generated from the two beamlines, it was determined that the SRC IRENI beamline was capable of producing higher-spatial-resolution distribution images in a shorter time than was achievable at the AS-IRM beamline, but that spectral noise levels per pixel were considerably lower on the AS-IRM beamline, resulting in more favourable data where the detection of weak absorbances is required. The data generated by the two complementary synchrotron IR methods on the chemical composition of cicada wings will be immensely useful in understanding their unusual properties with a view to reproducing their characteristics in, for example, industry applications.

Biophysical model of bacterial cell interactions with nanopatterned cicada wing surfaces.

The nanopattern on the surface of Clanger cicada (Psaltoda claripennis) wings represents the first example of a new class of biomaterials that can kill bacteria on contact based solely on their physical surface structure. The wings provide a model for the development of novel functional surfaces that possess an increased resistance to bacterial contamination and infection. We propose a biophysical model of the interactions between bacterial cells and cicada wing surface structures, and show that mechanical properties, in particular cell rigidity, are key factors in determining bacterial resistance/sensitivity to the bactericidal nature of the wing surface. We confirmed this experimentally by decreasing the rigidity of surface-resistant strains through microwave irradiation of the cells, which renders them susceptible to the wing effects. Our findings demonstrate the potential benefits of incorporating cicada wing nanopatterns into the design of antibacterial nanomaterials.

Selective bactericidal activity of nanopatterned superhydrophobic cicada Psaltoda claripennis wing surfaces.

The nanopattern on the surface of Clanger cicada (Psaltoda claripennis) wings represents the first example of a new class of biomaterials that can kill bacteria on contact based solely on its physical surface structure. As such, they provide a model for the development of novel functional surfaces that possess an increased resistance to bacterial contamination and infection. Their effectiveness against a wide spectrum of bacteria, however, is yet to be established. Here, the bactericidal properties of the wings were tested against several bacterial species, possessing a range of combinations of morphology and cell wall type. The tested species were primarily pathogens, and included Bacillus subtilis, Branhamella catarrhalis, Escherichia coli, Planococcus maritimus, Pseudomonas aeruginosa, Pseudomonas fluorescens, and Staphylococcus aureus. The wings were found to consistently kill Gram-negative cells (i.e., B. catarrhalis, E. coli, P. aeruginosa, and P. fluorescens), while Gram-positive cells (B. subtilis, P. maritimus, and S. aureus) remained resistant. The morphology of the cells did not appear to play any role in determining cell susceptibility. The bactericidal activity of the wing was also found to be quite efficient; 6.1 ± 1.5 × 10(6) P. aeruginosa cells in suspension were inactivated per square centimeter of wing surface after 30-min incubation. These findings demonstrate the potential for the development of selective bactericidal surfaces incorporating cicada wing nanopatterns into the design.

Spatial variations and temporal metastability of the self-cleaning and superhydrophobic properties of damselfly wings.

Self-cleaning surfaces found in nature show great potential for application in many fields, ranging from industry to medicine. The ability for a surface to self-clean is intimately related to the wetting properties of the surface; for a surface to possess self-cleaning ability it must exhibit extremely high water contact angles and low water adhesion. While investigating the self-cleaning properties of damselfly wings, significant spatial variations in surface wettability were observed. Within an area of 100 µm × 100 µm of the wing surface the water contact angle was found to vary up to 17.8°, while remaining consistently superhydrophobic. The contributions of both surface chemistry and topography to the hydrophobicity of the wings were assessed in an effort to explain these variations. Synchrotron-sourced Fourier-transform infrared microspectroscopy revealed that some of the major components of the wing were aliphatic hydrocarbons and esters, which are attributable to epicuticular lipids. The wing topography, as determined by optical profilometry and atomic force microscopy (AFM), also showed only minor levels of heterogeneity arising from irregular ordering of surface nanostructures. The measured contact angle of a single droplet of water was also found to decrease over time as it evaporated, reaching a minimum of 107°. This is well below the threshold value for superhydrophobicity (i.e., 150°), demonstrating that when the surface is in contact with water for a prolonged period, the damselfly wings lose their superhydrophobicity and subsequently their ability to self-clean. This decrease in hydrophobicity over time can be attributed to the surface undergoing a transition from the Cassie-Baxter wettability state toward the Wenzel wettability state.

New biomimetic directions in regenerative ophthalmology.

One of the most complete and permanent ways of treating many causes of visual impairment and blindness is to replace the entire affected tissue with pre-cultured ocular tissues supported and maintained on biomaterial frameworks. One direction towards enhancing ocular tissue regeneration on biomaterials, in the laboratory is by applying biomimicry. Specifically to engineer biomaterials with important functional elements of the native extracellular matrices, such as topography, that support and organise cells into coherent tissues. Further problems in regenerative ophthalmology can be potentially solved through application of biomimicry. They include, more efficient ways of moving and transplanting cultivated tissues into correct therapeutic locations inside the eye and scar-less, non-destructive healing of surgical incisions and wounds, to repair structural integrity of tissues at the ocular surface. Two examples are given to show this potential for redeveloping an ocular epithelium onto a nanostructured insect wing surface and producing an origami membrane modelled on deployable structures in nature. Efforts to harness natural innovation will eventually provide unique designs and structures that cannot for now be made synthetically, for regeneration of clinically acceptable ocular tissues.