RESUMO
Understanding the photodynamics of S-nitroso-thiol (RSNO), an effective NO transporter in biological systems, is essential for its photochemical applications. S-nitroso-mercaptoethanol (MceSNO), a simple water-soluble RSNO, facilitates high-level quantum calculations. We investigated the photoexcitation dynamics of MceSNO in an aqueous solution, focusing on NO dissociation, recombination, and linkage isomerization using quantum calculations and femtosecond infrared spectroscopy. Upon excitation at 320 nm, MceSNO rapidly dissociates into NO and MceS radicals. Approximately 31 ± 3% of MceS reacts with unexcited MceSNO molecules, forming MceSSMce and releasing additional NO. The remaining MceS undergoes geminate recombination with NO, forming either MceSNO (41 ± 4%) or MceSON (28 ± 3%), the latter being a sulfur-ON linkage isomer observed for the first time in a room-temperature solution. MceSON isomerizes back to MceSNO in 470 ± 30 ps. The formation mechanism of MceSON was verified through a potential energy surface constructed at the CASPT2D(16,11)/cc-pVTZ level. The isomerization barrier was determined to be 3.3 ± 1.2 kcal/mol in water.
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In this study, we developed a substrate-independent initiator film that can undergo surface-initiated polymerization to form an antifouling brush. Inspired by the melanogenesis found in nature, we synthesized a tyrosine-conjugated bromide initiator (Tyr-Br) that contains phenolic amine groups as the dormant coating precursor and α-bromoisobutyryl groups as the initiator. The resultant Tyr-Br was stable under ambient air conditions and underwent melanin-like oxidation only in the presence of tyrosinase to form an initiator film on various substrates. Subsequently, an antifouling polymer brush was formed using air-tolerant activators regenerated by electron transfer for atom transfer radical polymerization (ARGET ATRP) of zwitterionic carboxybetaine. The entire surface coating procedure, including the initiator layer formation and ARGET ATRP, occurred under aqueous conditions and did not require organic solvents or chemical oxidants. Therefore, antifouling polymer brushes can be feasibly formed not only on experimentally preferred substrates (e.g., Au, SiO2, and TiO2) but also on polymeric substrates such as poly(ethylene terephthalate) (PET), cyclic olefin copolymer (COC), and nylon.
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Inspired by the melanogenesis occurring in nature, we report tyrosinase-mediated antifouling surface coating by synthesizing a tyrosine-conjugated sulfobetaine derivative (Tyr-SB). Synthetic Tyr-SB contains zwitterionic sulfobetaine and tyrosine, whose phenolic amine group acts as a dormant coating precursor. In contrast to catecholamine derivatives, tyrosine derivatives are stable against auto-oxidation and are enzymatically oxidized only in the presence of tyrosinase to initiate melanin-like oxidation. When the surface of interest was applied during the course of Tyr-SB oxidation, a superhydrophilic poly(Tyr-SB) film was coated on the surfaces, thereby showing antifouling performance against proteins or adherent cells. Because the oxidation of Tyr-SB occurred under mild aqueous conditions (pH 6-7) without the use of any chemical oxidants, such as sodium periodate or ammonium persulfate, we anticipate that the coating method described herein will serve as a biocompatible tool in the field of biosensors, cell surface engineering, and medical devices, whose interfaces differ in chemistry.
Assuntos
Incrustação Biológica , Monofenol Mono-Oxigenase , Betaína/análogos & derivados , Incrustação Biológica/prevenção & controle , Catecolaminas , Melaninas , Oxidantes , TirosinaRESUMO
Dopamine (DA) surface chemistry has received significant attention because of its applicability in a wide range of research fields and the ability to graft functional molecules onto numerous solid surfaces. Various DA derivatives have been newly synthesized to identify key factors affecting the coating efficiency and to advance the coating system development. The oxidation of catechol into quinone followed by internal cyclization via the nucleophilic attack of primary amine is crucial for DA-based surface coating. Thus, it is expected that the amine group's nucleophilicity control directly affects the coating efficiency. However, it has not been systematically investigated, and most studies have been conducted with the focus on the transformation of amines into amides, despite such approaches decreasing the coating efficiency; the nitrogen in amides is less nucleophilic than that in free amines. In this study, we investigated the effect of N-alkylation on dopamine surface chemistry. N,N-Dimethyldopamine (DMDA) was newly synthesized, and the coating efficiency was systematically compared with DA and N-methyldopamine (MDA). DA N-monomethylation improved the coating rate by increasing the nitrogen nucleophilicity, whereas N,N-dimethylation dramatically decreased the DA surface coating property. In addition, MDA remained capable of universal surface coating and secondary reactions using the surface catechols. This study provides opportunities for developing coating materials with advanced functions and an improved coating rate.
Assuntos
Aminas , Dopamina , Amidas , Aminas/química , Dopamina/química , Metilação , Nitrogênio , Propriedades de SuperfícieRESUMO
A tyrosine-based azido derivative (TBAD) that permits both substrate-independent surface coating and clickable film functionalization by mimicking natural melanogenesis is synthesized here. In contrast to catechol derivatives, which are generally susceptible to oxidation by air under ambient conditions, the monophenol-based TBAD remains stable under alkaline and neutral conditions and is activated to oxidized quinone in situ by tyrosinase to initiate melanin-like polymerization. The resulting poly(TBAD) film can be formed on various substrates including noble metals, metal oxides, and synthetic polymers, which can undergo click reaction with terminal alkyne moieties on the entire surface or a specific region through Cu(I)-catalyzed azide-alkyne cycloaddition. The enzyme-mediated coating can rapidly form thin films (≈10 nm) and produce a uniform film morphology, which are important aspects in surface chemistry. This on-demand, clickable coating may become a significant tool for bioconjugation, soft lithography, and labeling techniques.
Assuntos
Química Click , Monofenol Mono-Oxigenase , Alcinos , Azidas , TirosinaRESUMO
Surface modification using alginic acid and its salt, alginate (Alg), has attracted much attention owing to its potential applications in various fields, including tissue engineering, drug delivery, antiplatelet surface preparation, and energy-storage technologies. In these applications, efficient immobilization of Alg on the solid surface is required because the delamination of the surface-bound Alg eventually leads to a significant decrease in its function. Therefore, much effort has been made to introduce Alg onto solid surfaces in a stable manner. Despite recent advances, existing methods for immobilizing Alg on surfaces have some limitations: (i) derivatization of Alg is typically also required and (ii) these methods only function under specific reaction conditions. Herein, we report a Zr(IV)-mediated strategy to immobilize Alg on solid surfaces. We demonstrate efficient Alg grafting onto carboxyl-, catechol-, polydopamine-, and tannic acid-functionalized surfaces via Zr(IV)-mediated cross-linking reactions. This strategy yields Alg multilayers that suppress fibroblast and platelet adhesion onto the solid surfaces. Furthermore, we show that the Alg multilayers can be selectively constructed on specific sites of solid surfaces. Given its ease of use and the wide selection of available carboxyl polymers, the current strategy is expected to be a useful tool for preparing functional polymer films for various applications.
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Herein, we report a degradable film that can be coated on various substrates by the codeposition of dopamine and cystamine. The thickness of the resulting film (pDC) varies depending on the initial ratio of dopamine/cystamine dissolved in a solution; the thickest film (ca. 60â nm) is obtained under optimized codeposition conditions. Selective degradation of pDC occurs in the presence of tris(2-carboxyethyl)phosphine (TCEP), the reaction kinetics of which are highly dependent on the TCEP concentration. For further application as a drug-delivery platform, doxorubicin can be loaded within the pDC film, which is released actively under film degradation in response to TCEP. We expect that the developed pDC film will be a useful tool for developing drug delivery cargo, antibacterial surface, and cell surface coating for various biomedical applications.
Assuntos
Antibacterianos/química , Cistamina/química , Dopamina/química , Doxorrubicina/química , Sistemas de Liberação de Medicamentos , Cinética , Estrutura Molecular , Propriedades de SuperfícieRESUMO
In this study, we developed a uniform initiator layer that can be formed on various surfaces, and formed site-selectively, for the subsequent antifouling polymer brush formation. Initially, metal-organic films composed of tannic acid (TA) and FeIII ions (TA-FeIII) were formed on various surfaces, followed by functionalization with an aryl azide-based initiator (ABI) under photoreaction. In particular, combination with a photolithographic technique enabled the presentation of initiators only on the intended region within a single-surface platform. A resultant initiator film (TF-ABI) was formed under mild reaction conditions and meets the uniformity and transparency requirements concurrently. Subsequently, we showed that TF-ABI can be further utilized to form a polymer brush by proceeding with surface-initiated polymerization using a zwitterionic monomer, namely, sulfobetaine acrylamide (SBAA). Instead of applying a classical, yet air-sensitive atom transfer radical polymerization (ATRP) technique, we utilized an activator regenerated by electron transfer (ARGET) ATRP under air conditions without a cumbersome deoxygenation step. Overall, our initiator layer allowed the antifouling poly(SBAA) brush to be used on various surfaces, and enabled their pattern generation.
RESUMO
The formation of a dense zwitterionic brush through surface-initiated atom transfer radical polymerization (SI-ATRP) is a typical graft-from approach used to achieve antifouling surfaces with high fidelity; however, their air-tightness may cause inconvenience to users. In this context, activator regenerated by electron transfer (ARGET) ATRP is emerging as an alternative surface-coating tool because limited amount of air is allowed to form a dense polymer brush. However, the degree of air tolerance that can ensure a thick polymer brush has not been clearly defined, limiting its practical usage under ambient-air conditions. In this study, we investigated the SI-ARGET ATRP of carboxybetaine (CB) by changing the air conditions, along with the air-related parameters, such as the concentration of the reducing agent, the volume of the polymerization solution (PS), or the solvent composition, and correlated their effects with the poly(CB) thickness. Based on the optimized reaction conditions, a poly(CB) brush with reliable thickness was feasibly formed even under open-air conditions without a degassing step. In addition, a microliter droplet (â¼100 µL) of PS was sufficient to proceed with the SI-ARGET ATRP for the covering of a poly(CB) brush on the surface area of interest. By applying an optimized SI-ARGET ATRP of CB, antifouling was feasibly achieved in the surface region of interest using an array to form a large surface area under fully exposed air conditions. In other words, optimized SI-ARGET ATRP enabled the formation of a thick poly(CB) brush on the surfaces of various dimensions under open-air conditions.
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Here, we report a simple yet effective surface-modification approach to imparting hydrophobic surfaces with superhydrophilicity using ultralow fouling/functionalizable carboxybetaine (CB) copolymers via a dip-coating technique. A new series of CB random copolymers with varying amphiphilicities were synthesized and coated on hydrophobic polypropylene (PP) and polystyrene (PS) surfaces. The nonfouling capability of each coating was screened by an enzyme-linked immunosorbent assay (ELISA) and further comprehensively assessed against 100% human serum by a Micro BCA protein assay kit. The random copolymer containing â¼30 mol % CB units showed superhydrophilicity with the highest air contact angle of more than 165° in DI water and the best nonfouling capability against 100% human blood serum. Surfaces of a 96-well plate coated with the optimal CB random copolymer had a significantly better nonfouling capability than those of a commercial 96-well plate with an ultralow attachment surface. The adhesion of mouse embryonic fibroblast cells (NIH3T3) was completely inhibited on surfaces coated with CB random copolymers. Furthermore, the optimal nonfouling CB copolymer surface was functionalized with an antigen via covalent bonding where its specific interactions with its antibody were verified. Thus, this CB random copolymer is capable of imparting both ultralow fouling and functionalizable capabilities to hydrophobic surfaces for blood-contacting devices.
Assuntos
Resinas Acrílicas/química , Incrustação Biológica/prevenção & controle , Compostos de Amônio Quaternário/química , Resinas Acrílicas/síntese química , Resinas Acrílicas/metabolismo , Adsorção , Animais , Proteínas Sanguíneas/metabolismo , Humanos , Camundongos , Células NIH 3T3 , Polipropilenos/química , Poliestirenos/química , Ligação Proteica , Compostos de Amônio Quaternário/síntese química , Compostos de Amônio Quaternário/metabolismoRESUMO
This work examines the development of primary neurons and astrocytes on thoroughly controlled functional groups. Negatively charged surfaces presenting carboxylate (COO-) or sulfonate (SO3-) groups prove beneficial to neuronal behavior, in spite of their supposed repulsive electrostatic interactions with cellular membranes. The adhesion and survival of primary hippocampal neurons on negatively charged surfaces are comparable to or slightly better than those on positively charged (poly-d-lysine-coated) surfaces, and neuritogenesis and neurite outgrowth are accelerated on COO- and SO3- surfaces. Moreover, such favorable influences of the negatively charged surfaces are only seen in neurons but not for astrocytes. Our results indicate that the in vitro developmental behavior of primary hippocampal neurons is sophisticatedly modulated by angstrom-sized differences in chemical structure or the charge density of the surface. We believe that this work provides new implications for understanding neuron-material interfaces as well as for establishing new ways to fabricate neuro-active surfaces.
Assuntos
Astrócitos/citologia , Astrócitos/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Hipocampo/citologia , Neurônios/citologia , Neurônios/efeitos dos fármacos , Animais , Ácidos Carboxílicos/química , Ácidos Carboxílicos/farmacologia , Membrana Celular/química , Membrana Celular/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Eletricidade Estática , Ácidos Sulfônicos/química , Ácidos Sulfônicos/farmacologia , Propriedades de SuperfícieRESUMO
Lymphocytes, such as T cells and natural killer (NK) cells, have therapeutic promise in adoptive cell transfer (ACT) therapy, where the cells are activated and expanded inâ vitro and then infused into a patient. However, the inâ vitro preservation of labile lymphocytes during transfer, manipulation, and storage has been one of the bottlenecks in the development and commercialization of therapeutic lymphocytes. Herein, we suggest a cell-in-shell (or artificial spore) strategy to enhance the cell viability in the practical settings, while maintaining biological activities for therapeutic efficacy. A durable titanium oxide (TiO2 ) shell is formed on individual Jurkat T cells, and the CD3 and other antigens on cell surfaces remain accessible to the antibodies. Interleukin-2 (IL-2) secretion is also not hampered by the shell formation. This work suggests a chemical toolbox for effectively preserving lymphocytes inâ vitro and developing the lymphocyte-based cancer immunotherapy.
Assuntos
Terapia Baseada em Transplante de Células e Tecidos , Imunoterapia , Neoplasias/terapia , Linfócitos T/efeitos dos fármacos , Titânio/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Humanos , Células Jurkat , Neoplasias/imunologia , Linfócitos T/citologia , Linfócitos T/imunologia , Titânio/químicaRESUMO
We achieved ultralow fouling on target surfaces by controlled polymerization of carboxybetaine under ambient conditions. The polymerization process for grafting polymer films onto the surfaces was carried out in air and did not require any deoxygenation step or specialized equipment. This method allows one to conveniently introduce a nonfouling polymer network onto large substrates.
RESUMO
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.
RESUMO
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.
Assuntos
Magnetismo , Saccharomyces cerevisiae/citologia , Ouro/farmacologia , Dióxido de Silício/farmacologia , Fatores de TempoRESUMO
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.
Assuntos
Células , NanoestruturasRESUMO
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.
Assuntos
Adesivos/química , Dissulfetos/química , Dopamina/química , Indóis/química , Nanotecnologia/métodos , Polímeros/química , Proteínas/química , Animais , Materiais Biocompatíveis/química , Bivalves , Soluções Tampão , Materiais Revestidos Biocompatíveis/química , Doxorrubicina/química , Sistemas de Liberação de Medicamentos , Glutationa/química , Levodopa/química , Espectroscopia de Infravermelho com Transformada de Fourier , Propriedades de Superfície , Engenharia Tecidual/métodos , Água/químicaRESUMO
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.
Assuntos
Proliferação de Células , Compostos Férricos , Protetores Solares , Taninos , Raios Ultravioleta , Animais , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/efeitos da radiação , Compostos Férricos/química , Compostos Férricos/farmacocinética , Compostos Férricos/farmacologia , Células HeLa , Humanos , Células Jurkat , Camundongos , Células NIH 3T3 , Protetores Solares/química , Protetores Solares/farmacocinética , Protetores Solares/farmacologia , Taninos/química , Taninos/farmacocinética , Taninos/farmacologiaRESUMO
We report a perfluoroaryl azide-based photoreaction for synthesizing functionalizable and nonbiofouling poly[oligo(ethylene glycol) methacrylate] (pOEGMA) films on a chemically inert COC substrate, and an estimation of a surface coverage of the antibody immobilized onto the surface with the immuno-gold nanoparticles. The processes were confirmed by water contact angle measurement, FT-IR spectroscopy, and FE-SEM. The strategy demonstrated in this work could be applied to functionalizations of other polymeric materials and determination of the binding capacity of analytes in biosensors and microfluidic devices.
Assuntos
Alcenos/química , Anticorpos/química , Ouro/química , Nanopartículas Metálicas/química , Metacrilatos/química , Nanoconjugados/química , Polietilenoglicóis/química , Adsorção , Animais , Anticorpos/imunologia , Imunoglobulina G/análise , Imunoglobulina G/imunologia , Teste de Materiais , Nanopartículas Metálicas/ultraestrutura , Camundongos , Nanoconjugados/ultraestrutura , Tamanho da Partícula , Propriedades de SuperfícieRESUMO
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.