Single-Cell Nanoencapsulation: From Passive to Active Shells.

Single-cell nanoencapsulation is an emerging field in cell-surface engineering, emphasizing the protection of living cells against external harmful stresses in vitro and in vivo. Inspired by the cryptobiotic state found in nature, cell-in-shell structures are formed, which are called artificial spores and which show suppression or retardation in cell growth and division and enhanced cell survival under harsh conditions. The property requirements of the shells suggested for realization of artificial spores, such as durability, permselectivity, degradability, and functionalizability, are demonstrated with various cytocompatible materials and processes. The first-generation shells in single-cell nanoencapsulation are passive in the operation mode, and do not biochemically regulate the cellular metabolism or activities. Recent advances indicate that the field has shifted further toward the formation of active shells. Such shells are intimately involved in the regulation and manipulation of biological processes. Not only endowing the cells with new properties that they do not possess in their native forms, active shells also regulate cellular metabolism and/or rewire biological pathways. Recent developments in shell formation for microbial and mammalian cells are discussed and an outlook on the field is given.

Publisher Correction: Antimicrobial spray nanocoating of supramolecular Fe(III)-tannic acid metal-organic coordination complex: applications to shoe insoles and fruits.

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Enzymatic film formation of nature-derived phenolic amines.

An enzyme-instructed method is developed for material-independent, cytocompatible coating of phenolic amines, inspired by melanogenesis found in nature. Tyrosinase-based film formation proceeds smoothly in an aqueous solution at neutral pH, and can use various phenolic amines including catecholamines, such as tyrosine, tyramine, dopamine, norepinephrine, and DOPA, as a coating precursor. Compared with polydopamine coating, the method is fast and efficient, and forms uniform films. Its high cytocompatibility is advantageously applied to cell-surface engineering, where chemically labile Jurkat cells are coated individually without any noticeable decrease in viability. Considering the huge potential of polyphenolic-based coatings, the strategy developed herein will provide an advanced tool for manipulating biological entities, including living cells, in biomedical and medicinal applications.

Antimicrobial spray nanocoating of supramolecular Fe(III)-tannic acid metal-organic coordination complex: applications to shoe insoles and fruits.

Numerous coating strategies are available to control the surface properties and confer new properties to substrates for applications in energy, environment, biosystems, etc., but most have the intrinsic limitations in the practical setting: (1) highly specific interactions between coating materials and target surfaces are required for stable and durable coating; (2) the coating of bulk substrates, such as fruits, is time-consuming or is not achievable in the conventional solution-based coating. In this respect, material-independent and rapid coating strategies are highly demanded. We demonstrate spray-assisted nanocoating of supramolecular metal-organic complexes of tannic acid and ferric ions. The spray coating developed is material-independent and extremely rapid (<5 sec), allowing for coating of commodity goods, such as shoe insoles and fruits, in the controlled fashion. For example, the spray-coated mandarin oranges and strawberries show significantly prolonged post-harvest shelf-life, suggesting practical potential in edible coating of perishable produce.

Artificial Spores: Immunoprotective Nanocoating of Red Blood Cells with Supramolecular Ferric Ion-Tannic Acid Complex.

The blood-type-mismatch problem, in addition to shortage of blood donation, in blood transfusion has prompted the researchers to develop universal blood that does not require blood typing. In this work, the "cell-in-shell" (i.e., artificial spore) approach is utilized to shield the immune-provoking epitopes on the surface of red blood cells (RBCs). Individual RBCs are successfully coated with supramolecular metal-organic coordination complex of ferric ion (FeIII) and tannic acid (TA). The use of isotonic saline (0.85% NaCl) is found to be critical in the formation of stable, reasonably thick (20 nm) shells on RBCs without any aggregation and hemolysis. The formed "RBC-in-shell" structures maintain their original shapes, and effectively attenuate the antibody-mediated agglutination. Moreover, the oxygen-carrying capability of RBCs is not deteriorated after shell formation. This work suggests a simple but fast method for generating immune-camouflaged RBCs, which would contribute to the development of universal blood.

Turning Diamagnetic Microbes into Multinary Micro-Magnets: Magnetophoresis and Spatio-Temporal Manipulation of Individual Living Cells.

Inspired by the biogenic magnetism found in certain organisms, such as magnetotactic bacteria, magnetic nanomaterials have been integrated into living cells for bioorthogonal, magnetic manipulation of the cells. However, magnetized cells have so far been reported to be only binary system (on/off) without any control of magnetization degree, limiting their applications typically to the simple accumulation or separation of cells as a whole. In this work, the magnetization degree is tightly controlled, leading to the generation of multiple subgroups of the magnetized cells, and each subgroup is manipulated independently from the other subgroups in the pool of heterogeneous cell-mixtures. This work will provide a strategic approach to tailor-made fabrication of magnetically functionalized living cells as micro-magnets, and open new vistas in biotechnological and biomedical applications, which highly demand the spatio-temporal manipulation of living cells.

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.

Artificial Spores: Cytocompatible Coating of Living Cells with Plant-Derived Pyrogallol.

Cell nanoencapsulation, generating cell-in-shell structures ("artificial spores"), provides a chemical toolbox for controlling the cellular behaviors and functional characteristics of individual cells. Among the shell materials studied so far, naturally occurring polyphenolic compounds, including polydopamine and tannic acid, have intensively been employed in cell-surface engineering, because their material-independent coating property eliminates an extra priming step for inducing subsequent shell formation. Albeit successful in generating cell-in-shell structures, the coating of polyphenolic compounds generally requires alkaline conditions and/or high salt conditions, which are not compatible with certain cell types. In this work, we demonstrate that the nanocoating of individual cells with a plant-derived phenolic compound, pyrogallol (1,2,3-trihydroxybenzene), occurs at mildly alkaline pH of 7.8 in an isotonic buffer. Three different cell types (anucleate, microbial, and mammalian cells) are coated with pyrogallol without noticeable decrease in cell viability. The protocol developed in this work could be applied to other polyphenolic compounds, and, considering the many polyphenols identified as a coating material, provides an advanced chemical tool in cell-surface engineering.

Comparative Study on Surface-Initiated ATRP and SI-ARGET ATRP of Oligo(Ethylene Glycol) Methacrylate on Gold.

Oligo(ethylene glycol) methacrylate (OEGMA) was polymerized from a polymerization initiator-presenting gold substrate by Activator ReGenerated by Electron Transfer Atom Transfer Radical Polymerization (ARGET ATRP) in water. Compared with the normal surface-initiated ATRP (SI-ATRP), SI-ARGET ATRP of OEGMA proceeded smoothly in the presence of air with L-ascorbic acid as a reducing agent and a CuBr2/2,2'-bipyridyl complex at the ppm level. In addition, SI-ARGET ATRP did not require the additional steps for removing a polymerization inhibitor from the OEGMA monomer and for deoxygenating the solvent. The ellipsometric measurements showed that the polymerized OEGMA (pOEGMA) films prepared by SI-ARGET ATRP were on average 10 times thicker than those prepared by normal SI-ATRP with the same monomer concentration and polymerization time.

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.

Cytoprotective alginate/polydopamine core/shell microcapsules in microbial encapsulation.

Chemical encapsulation of microbes in threedimensional polymeric microcapsules promises various applications, such as cell therapy and biosensors, and provides a basic platform for studying microbial communications. However, the cytoprotection of microbes in the microcapsules against external aggressors has been a major challenge in the field of microbial microencapsulation, because ionotropic hydrogels widely used for microencapsulation swell uncontrollably, and are physicochemically labile. Herein, we developed a simple polydopamine coating for obtaining cytoprotective capability of the alginate capsule that encapsulated Saccharomyces cerevisiae. The resulting alginate/ polydopamine core/shell capsule was mechanically tough, prevented gel swelling and cell leakage, and increased resistance against enzymatic attack and UV-C irradiation. We believe that this multifunctional core/shell structure will provide a practical tool for manipulating microorganisms inside the microcapsules.

Bioinspired, cysteamine-catalyzed co-silicification of (1H, 1H, 2H, 2HPerfluorooctyl) triethoxysilane and tetraethyl orthosilicate: formation of superhydrophobic surfaces.

Bioinspired silicification attracts a great deal of interest because of its physiologically relevant, mild conditions for hydrolysis and condensation of silica precursors, which makes the bioinspired approach superior to the conventional sol­gel process, particularly when dealing with biological entities. However, the morphological control of silica structures with incorporation of functional groups in the bioinspired silicilication has been unexplored. In this work, we co-silicificated (1H, 1H, 2H, 2H-perfluorooctyl)triethoxysilane and tetraethyl orthosilicate to investigate the morphological evolution of fluorinated silica structures in the cetyltrimethylammonium bromide-mediated, cysteamine-catalyzed silicification. The generated micrometer-long wormlike and spherical silica structures display superhydrophobicity after film formation. Interestingly, the measurement of dynamic water contact angles shows that the morphological difference leads to a different wetting state, either the self-cleaning or the pinning state of the superhydrophobic surface.