Search

Nature has developed a fascinating strategy of cryptobiosis ("secret life") for counteracting the stressful, and often lethal, environmental conditions that fluctuate sporadically over time. For example, certain bacteria sporulate to transform from a metabolically active, vegetative state to an ametabolic endospore state. The bacterial endospores, encased within tough biomolecular shells, withstand the extremes of harmful stressors, such as radiation, desiccation, and malnutrition, for extended periods of time and return to a vegetative state by breaking their protective shells apart when their environment becomes hospitable for living. Certain ciliates and even higher organisms, for example, tardigrades, and others are also found to adopt a cryptobiotic strategy for survival. A common feature of cryptobiosis is the structural presence of tough sheaths on cellular structures. However, most cells and cellular assemblies are not "spore-forming" and are vulnerable to the outside threats. In particular, mammalian cells, enclosed with labile lipid bilayers, are highly susceptible to in vitro conditions in the laboratory and daily life settings, making manipulation and preservation difficult outside of specialized conditions. The instability of living cells has been a main bottleneck to the advanced development of cell-based applications, such as cell therapy and cell-based sensors. A judicious question arises: can cellular tolerance against harmful stresses be enhanced by simply forming cell-in-shell hybrid structures? Experimental results suggest that the answer is yes. A micrometer-sized "Iron Man" can be generated by chemically forming an ultrathin (100 nm) but durable shell on a "non-spore-forming" cell. Since the report on silica nanoencapsulation of yeast cells, in which cytoprotective yeast-in-silica hybrids were formed, several synthetic strategies have been developed to encapsulate individual cells in a cytocompatible fashion, mimicking the cryptobiotic cell-in-shell structures found in nature, for example, bacterial endospores. Bioinspired silicification and phenolics-based coatings are, so far, the main approaches to the formation of cytoprotective cell-in-shell hybrids, because they ensure cell viability during encapsulations and also generate durable nanoshells on cell surfaces. The resulting cell-in-shell hybrids extrinsically possess enhanced resistance to external aggressors, and more intriguingly, the encapsulation alters their metabolic activity, exemplified by retarded or suppressed cell cycle progression. In addition, recent developments in the field have further advanced the synthetic tools available to the stage of chemical sporulation and germination of mammalian cells, where cytoprotective shells are formed on labile mammalian cells and broken apart on demand. For example, individual HeLa cells are coated with a metal-organic complex of ferric ion and tannic acid, and cellular adherence and proliferation are controlled by the programmed shell formation and degradation. Based on these demonstrations, the (degradable) cell-in-shell hybrids are anticipated to find their applications in various biomedical and bionanotechnological areas, such as cytotherapeutics, high-throughput screening, sensors, and biocatalysis, as well as providing a versatile research platform for single-cell biology.

In this Opinion we discuss the development of artificial spores and their maturation as an independent field of research. The robust cell-in-shell structures have displayed unprecedented characteristics, which include the retardation of cell division and extensive cytoprotective capabilities that encompass exposure to osmotic pressure, shear force, heat, UV radiation, and lytic enzymes. Additionally, the nanothin shells act as highly versatile scaffolds for chemical functionalization to equip cells for implementation in tissue engineering, biosensors, cell therapy, or other biotechnological applications. We also explore the future direction of this emerging field and dictate that the next phase of research should focus on attaining more intricate engineering to achieve stimulus-responsive shell-degradation, multilayer casings with orthogonal functions, and the encapsulation of multiple cells for multicellular artificial spores.

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.

E-mail: sunghoyang@knue.ac.krReceived June 16, 2014, Accepted July 2, 2014Key Words : Silica, Thin film, Histidine, Self-assembled monolayer, SilicificationThere exists a great deal of interest in silica thin films,which potentially allow us to develop various applicationsincluding heterogeneous catalysis

In this work, we report that high-density, vertically grown silicon nanowires (vg-SiNWs) direct a new in vitro developmental pathway of primary hippocampal neurons. Neurons on vg-SiNWs formed a single, extremely elongated major neurite earlier than minor neurites, which led to accelerated polarization. Additionally, the development of lamellipodia, which generally occurs on 2D culture coverslips, was absent on vg-SiNWs. The results indicate that surface topography is an important factor that influences neuronal development and also provide implications for the role of topography in neuronal development in vivo.

Although the programmed degradation of biocompatible films finds applications in various fields including biomedical and bionanotechnological areas, coating methods have generally been limited to be substrate-specific, not applicable to any kinds of substrates. In this paper, we report a dopamine derivative, which allows for both universal coating of various substrates and stimuli-responsive film degradation, inspired by mussel-adhesive proteins. Two dopamine moieties are linked together by the disulfide bond, the cleavage of which enables the programmed film degradation. Mechanistic analysis of the degradable films indicates that the initial cleavage of the disulfide linkage causes rapid uptake of water molecules, hydrating the films, which leads to rapid degradation. Our substrate-independent coating of degradable films provides an advanced tool for drug delivery systems, tissue engineering, and anti-fouling strategies.

The design of stimuli-responsive self-assembled molecular systems capable of undergoing mechanical work is one of the most important challenges in synthetic chemistry and materials science. Here we report that foldectures, that is, self-assembled molecular architectures of β-peptide foldamers, uniformly align with respect to an applied static magnetic field, and also show instantaneous orientational motion in a dynamic magnetic field. This response is explained by the amplified anisotropy of the diamagnetic susceptibilities as a result of the well-ordered molecular packing of the foldectures. In addition, the motions of foldectures at the microscale can be translated into magnetotactic behaviour at the macroscopic scale in a way reminiscent to that of magnetosomes in magnetotactic bacteria. This study will provide significant inspiration for designing the next generation of biocompatible peptide-based molecular machines with applications in biological systems., Controlling organic materials with magnetic fields in a dynamic fashion is a challenging task. Here, the authors show that synthetic ß-peptide foldamers can be rotated at will under a dynamic magnetic field and that this can be extended to macroscopic scale objects containing these materials.

Formation of both mechanically durable and programmably degradable layer-by-layer (LbL) films in a biocompatible fashion has potential applications in cell therapy, tissue engineering, and drug-delivery systems, where the films are interfaced with living cells. In this work, we developed a simple but versatile method for generating in situ cross-linked and responsively degradable LbL films, based on the thiol–exchange reaction, under highly cytocompatible conditions (aqueous solution at pH 7.4 and room temperature). The cytocompatibility of the processes was confirmed by coating individual yeast cells with the cross-linked LbL films and breaking the films on demand, while maintaining the cell viability. In addition, the processes were applied to the controlled release of an anticancer drug in the HeLa cells.

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".

In this chapter, we describe methods for encapsulating individual cells within inorganic nanoshells. Inspired by the biomineralization that occurs in nature, living cells have been encapsulated within various inorganic shells made of gold, calcium phosphate, calcium carbonate, silica, or titania. The viability and metabolic activities of the cells were ensured by the careful selection of biocompatible materials and processes. Importantly, inorganic shells have been useful for enhancing long-term cell viability, controlling cell division, protecting the cytoplasm against foreign aggression, and functionalizing cell surfaces. This chapter focuses on the chemical reactions used to generate inorganic shells, categorized as direct reduction, in situ crystallization, and in situ condensation, and on the artificial control of the cellular behaviors conferred by the inorganic shells. The future prospects in the field of cell encapsulation within inorganic materials are also discussed.

The chronological progresses in single-cell nanocoating are described. The historical developments in the field are divided into biotemplating, cytocompatible nanocoating, and cells in nano-nutshells, depending on the main research focuses. Each subfield is discussed in conjunction with the others, regarding how and why to manipulate living cells by nanocoating at the single-cell level.

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.