RESUMEN
Thermoresponsive self-assembled nanogels were conveniently prepared by cholesterol end-capped poly(N-isopropylacrylamide) (PNIPAM) in water. Both cholesterol end-capped PNIPAMs (telelchelic cholesterol PNIPAM, tCH-PNIPAM) formed flower-like nanogels by the self-assembling of four to five polymer chains with multiple domains of cholesterol in water at 20 °C. Meanwhile, one end-group cholesterol-capped PNIPAM (semitelechelic cholesterol PNIPAM, stCH-PNIPAM) was also formed as a nanogel by the self-assembling of 15-20 polymer chains with 3 to 4 cholesterol domains. The hydrophobic cholesterol domains of tCH-PNIPAM nanogels were maintained above the lower critical solution temperature (LCST) of PNIPAM (>32 °C). Differently, the hydrophobic domains of stCH-PNIPAM were disrupted by cholesterol-free PNIPAM chain ends and formed large mesoglobules above the LCST. These transition controls of hydrophilic end-capped smart polymers may open new methodologies to design thermoresponsive nanosystems.
Asunto(s)
Calor , Agua , Resinas Acrílicas , Colesterol/química , Nanogeles , Polímeros/química , TemperaturaRESUMEN
Thermoresponsive polymers that possess both UCST- and LCST-like behaviors have generally been designed using diblock copolymers that are mostly composed of an LCST-like polymer and a UCST polymer. Herein, we prepared an LCST-UCST-type polymer composed of UCST-like thermoresponsive zwitterionic sulfabetaine methacrylate and nonthermoresponsive PEG methacrylate, ZB-PEG, by reversible addition-fragmentation chain transfer (RAFT) copolymerization. By adjusting the PEG composition, ZB-PEG formed a mesoglobule, a microglobule, and the dissociated states in phosphate-buffered saline (PBS). These states were found to be reversible via temperature control. Moreover, this behavior showed high reversibility and succeeded in stabilizing horseradish peroxidase (HRP) in the dilute condition. Such thermoresponsive ZB-PEG can be applied over a wide range of applications in biotechnology and other fields.
RESUMEN
Understanding the interactions of soft nanomatters with cell membranes is particularly important for research into nanocarrier-based drug delivery systems, cell engineering, and subcellular imaging. Most nanoparticles, vesicles, micelles, and polymeric aggregates are internalized into endosomes and, eventually, lysosomes in the cytosol because of energy-dependent endocytic processes. Endocytic uptake substantially limits the access to the cytoplasm where a cargo agent acts. Bypassing the endocytic pathways by direct penetration into plasma membrane barriers would enhance the efficacy of nanomedicines. Some zwitterionic polymer nanoaggregates have been shown to permeate into the cell interior in an energy-independent manner. We have elucidated this phenomenon by observing changes in the biomembrane barrier functions against protons as the smallest indicator and have used these results to further design and develop poly(betaines). In this work, we investigated the translocation mechanisms for a series of zwitterionic poly(methacrylamide) and poly(methacrylate) species bearing a pyridinium propane sulfonate moiety in the monomers. Minor differences in the monomer structures and functional groups were observed to have dramatic effects on the interaction with plasma membranes during translocation. The ability to cross the plasma membrane involves a balance among the betaine dipole-dipole interaction, NH-π interaction, π-π interaction, cation-π interaction, and amide hydrogen bonding. We found that the cell-penetrating polysulfobetaines had limited or no detrimental effect on cell proliferation. Our findings enhance the opportunity to design and synthesize soft nanomatters for cell manipulations by passing across biomembrane partitions.
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Betaína , Polímeros , Betaína/análogos & derivados , Membrana Celular , MicelasRESUMEN
Three-dimensional cell aggregates (spheroids) are becoming a research focus because their construction is similar to that in vivo microenvironments, enabling the acceleration of drug discovery and reducing the need for animal tests, and other advantages. However, the delivery of drugs to the inside of spheroids is time-consuming and has low efficiency. In this study, we selected a sulfobetaine copolymer that translocates to the cell membrane in monolayer cultured cells as a nanocarrier of anticancer drugs. Doxorubicin (Dox) and 17-demethoxy-17-allylamino geldanamycin (17AAG) were modified to the copolymer of sulfobetaine methacrylate and poly(ethylene glycol) methacrylate, P(SB-PEG), and added to glioblastoma A-172 cell spheroids. Dox-P(SB-PEG) showed fast permeation into A-172 spheroids, and the fluorescence in cells was observed in the center area of the spheroids within 1 h of polymer addition. Conversely, only the outer one to two cell layers of spheroids were observed when Dox was added to the spheroids. Dox-P(SB-PEG) in A-172 spheroids was localized in the mitochondria of each cell and exhibited comparable drug efficacy to that of Dox in growth inhibition assays of A-172 spheroids. Moreover, approximately 10-fold higher drug efficacy in growth inhibition and invasion of A-172 spheroids was found using 17AAG-P(SB-PEG). Conjugating anticancer drugs with P(SB-PEG) is a promising strategy to enhance drug permeation and efficacy against spheroid cells.
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Antineoplásicos , Neoplasias Encefálicas , Glioblastoma , Polietilenglicoles , Animales , Antineoplásicos/farmacología , Betaína/análogos & derivados , Neoplasias Encefálicas/tratamiento farmacológico , Línea Celular Tumoral , Doxorrubicina/farmacología , Portadores de Fármacos , Glioblastoma/tratamiento farmacológico , Polímeros , Microambiente TumoralRESUMEN
Silicon dioxide (SiO2) nanoparticles (NPs) and titanium dioxide (TiO2) NPs are the most widely used inorganic nanomaterials. Although the individual toxicities of SiO2 and TiO2 NPs have been extensively studied, the combined toxicity of these NPs is much less understood. In this study, we observed unexpected and drastic activation of the caspase-1 inflammasome and production of IL-1ß in mouse bone marrow-derived macrophages stimulated simultaneously with SiO2 and TiO2 NPs at concentrations at which these NPs individually do not cause macrophage activation. Consistent with this, marked lung inflammation was observed in mice treated intratracheally with both SiO2 and TiO2 NPs. In macrophages, SiO2 NPs localized in lysosomes and TiO2 NPs did not; while only TiO2 NPs produced ROS, suggesting that these NPs induce distinct cellular damage leading to caspase-1 inflammasome activation. Intriguingly, dynamic light scattering measurements revealed that, although individual SiO2 and TiO2 NPs immediately aggregated to be micrometer size, the mixture of these NPs formed a stable and relatively monodisperse complex with a size of ~250 nm in the presence of divalent cations. Taken together, these results suggest that SiO2 and TiO2 NPs synergistically induce macrophage inflammatory responses and subsequent lung inflammation. Thus, we propose that it is important to assess the synergistic toxicity of various combinations of nanomaterials.
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Interleucina-1beta/metabolismo , Macrófagos/efectos de los fármacos , Nanopartículas/toxicidad , Neumonía/inducido químicamente , Dióxido de Silicio/toxicidad , Titanio/toxicidad , Animales , Modelos Animales de Enfermedad , Sinergismo Farmacológico , Femenino , Inflamasomas/inmunología , Inflamación , Interleucina-1beta/inmunología , Macrófagos/inmunología , Ratones Endogámicos C57BL , Tamaño de la Partícula , Neumonía/inmunología , Dióxido de Silicio/farmacocinética , Propiedades de Superficie , Titanio/farmacocinéticaRESUMEN
The majority of nanoparticles designed for cellular delivery of drugs and imaging agents enter the cell via endocytotic pathways leading to their entrapment in endosomes that present a robust barrier to further trafficking of the nanoparticles within the cells. A few materials, such as the cell penetrating peptides (CPPs), are known to enter cells not only via endocytosis, but also via translocation through the cell membrane into the cytoplasm, successfully bypassing the endosomes. We report here that random copolymers of 3-dimethyl(methacryloyloxyethyl)ammonium propanesulfonate and poly(ethylene glycol) methacrylate, p(DMAPS-ran-PEGMA), are internalized in cells primarily via translocation through the cell membrane rather than endocytosis. The properties of the polymers and their modes of uptake were investigated systematically by dynamic light scattering, confocal fluorescence microscopy, and flow cytometry. Using specific inhibitors of the cellular uptake machinery in a human cervical carcinoma cell line (HeLa), we show that these nontoxic synthetic polyzwitterions exist in cell media as self-assembled nanospheres that unravel as they adsorb on the plasma membrane and translocate through it. Conjugates of p(DMAPS-ran-PEGMA) with rhodamine B were delivered selectively to the mitochondria, whereas doxorubicin (Dox)-p(DMAPS-ran-PEGMA) conjugates were accumulated in both the nucleus and the mitochondria, effectively inducing apoptosis in HeLa cells. These findings suggest that the noncytotoxic and readily synthesized p(DMAPS-ran-PEGMA) can find applications as bioimaging tools and drug nanocarriers.
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Doxorrubicina/metabolismo , Portadores de Fármacos/química , Metacrilatos/metabolismo , Nanosferas/química , Polietilenglicoles/metabolismo , Compuestos de Amonio Cuaternario/metabolismo , Rodaminas/metabolismo , Animales , Transporte Biológico/fisiología , Células CHO , Línea Celular Tumoral , Núcleo Celular/efectos de los fármacos , Péptidos de Penetración Celular , Cricetulus , Dispersión Dinámica de Luz , Citometría de Flujo , Células HL-60 , Células HeLa , Células Hep G2 , Humanos , Metacrilatos/química , Microscopía Confocal , Mitocondrias/efectos de los fármacos , Neoplasias/tratamiento farmacológico , Polietilenglicoles/química , Compuestos de Amonio Cuaternario/químicaRESUMEN
We investigate the effects of urea and its derivatives on the ATPase activity and on the in vitro motility of chicken skeletal muscle actomyosin. Mg-ATPase rate of myosin subfragment-1 (S1) is increased by 4-fold by 0.3M 1,3-diethylurea (DEU), but it is unaffected by urea, thiourea, and 1,3-dimethylurea at ≤1M concentration. Thus, we further examine the effects of DEU in comparison to those of urea as reference. In in vitro motility assay, we find that in the presence of 0.3M DEU, the sliding speeds of actin filaments driven by myosin and heavy meromyosin (HMM) are significantly decreased to 1/16 and 1/6.6, respectively, compared with the controls. However, the measurement of the actin-activated ATPase activity of HMM shows that the maximal rate, Vmax, is almost unchanged with DEU. Thus, the myosin-driven sliding motility of actin filaments is significantly impeded in the presence of 0.3M DEU, whereas the cyclic interaction of myosin with F-actin occurs during the ATP turnover, the rate of which is close to that without DEU. In contrast to DEU, 0.3M urea exhibits only modest effects on both actin-activated ATPase and sliding motility of actomyosin. Thus, DEU has the effect of uncoupling the sliding motility of actomyosin from its ATP turnover.
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Actinas/química , Actomiosina/química , Adenosina Trifosfato/química , Movimiento , Subfragmentos de Miosina/química , Miosinas del Músculo Esquelético/química , Actinas/metabolismo , Actomiosina/metabolismo , Adenosina Trifosfato/metabolismo , Animales , Pollos , Subfragmentos de Miosina/metabolismo , Miosinas del Músculo Esquelético/metabolismoRESUMEN
A double hydrophilic block copolymer, poly(ethylene glycol)-poly(3-dimethyl (methacryloyloxyethyl) ammonium propane sulfonate) (PEG-SB), is synthesized by reversible addition-fragmentation transfer (RAFT) polymerization using PEG methyl ether (4-cyano-4-pentanoate dodecyl trithiocarbonate) as a chain transfer agent. PEG-SB forms multi-layered microspheres with dipole-dipole interactions of the SB side chains as the driving force. The PEG-SB polymers show an upper critical solution temperature (UCST) and the UCST is controllable by the polymerization degree. The PEG-SB microspheres are dissociated above the UCST and then monodispersed microspheres (â¼1 µm) are obtained when the solution temperature is decreased below the UCST again. The disassociation/association of the microspheres is also controllable using the concentration of NaCl. These multi-responsive microspheres could be a powerful tool in the field of nano-biotechnology.
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Microesferas , Polímeros/química , Agua/química , Biotecnología , Microscopía Electrónica de Transmisión , NanotecnologíaRESUMEN
Sandwich culture systems are techniques that cultivate cells by sandwiching them between the top and bottom substrates. Since the substrates can be separated, the system is expected to be applied to the construct layering of patterned cells and to the isolation of stacked cells. In this study, we prepared hydrogels composed of zwitterionic sulfabetaine polymers, poly[2-(2-(methacryloyloxyethyl)dimethylammonio)ethyl-1-sulfate] (PZBMA). The ZBMA homopolymers have been shown to form aggregates in aqueous solutions due to their intermolecular interactions. The water content of the PZBMA hydrogels in water was â¼70% regardless of N,N'-methylenebis(acrylamide), BIS, content as the cross-linker. The results indicated that the intermolecular interaction contributed more to the swelling behaviors than the chemical cross-linker. However, PZBMA hydrogels with 0.1 mol % BIS showed not only high elongation (â¼850%) properties but also high adhesiveness and self-healing properties. When this PZBMA hydrogel was impregnated with collagen and subjected to sandwich culture using Madin-Darby canine kidney (MDCK) cells, a three-dimensional morphology of MDCK cell aggregates was constructed. Such a sulfabetaine hydrogel is expected to be developed for regenerative medicine.
RESUMEN
In this study, we prepared a new associating polymer, ChMaPLL, by the substitution of the poly(L-lysine) moiety with oligosaccharide amylose primer and cholesterol. ChMaPLL formed positively charged polypeptide nanogels (~50 nm) via self-assembly in water. The nanogels showed a secondary structural transition to an α-helix structure induced by poly(L-lysine) in response to an increase in pH. Oligosaccharides of the nanogels reacted with the phosphorylase a enzyme. Amylose-conjugated nanogels were obtained by enzymatic polymerization. The elongation of the saccharide chain shielded the positive charge of the nanogels. The multiresponsive polysaccharide-polypeptide hybrid nanogels might prove to be useful in the areas of biotechnology and biomedicine.
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Amilosa/química , Biopolímeros/química , Colesterol/química , Cabello/química , Nanoestructuras , Polilisina/química , Polisacáridos/química , Dicroismo Circular , Colorantes Fluorescentes , Humanos , Tamaño de la PartículaRESUMEN
A self-assembled nanogel, derived from an acid-labile cholesteryl-modified pullulan (acL-CHP), was prepared by grafting vinyl ether-cholesterol substituents onto a 100 kD pullulan main chain polymer backbone. Stable nanogels are formed by acL-CHP self-assemblies at neutral pH. The hydrodynamic radius of the nanogels, observed to be 26.5 ± 5.1 nm at pH 7.0, increased by ~135% upon acidification of the solution to pH 4.0. SEC analysis of the acL-CHP nanogel at pH 4.0 showed that the grafts were nearly 80% degraded after 24 h, whereas little or no degradation was observed over the same time period for a pH stable analog (acS-CHP) at pH 4.0 or the acL-CHP at pH 7.0. Complexation of BSA with the acL-CHP nanogel was observed at pH 7.0 with subsequent release of the protein upon acidification. These findings suggest that stimuli-responsive, self-assembled nanogels can release protein cargo in a manner that is controlled by the degradation rate of the cholesterol-pullulan grafting moiety.
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Colesterol/química , Sistemas de Liberación de Medicamentos/métodos , Glucanos/química , Polietilenglicoles/química , Polietileneimina/química , Albúmina Sérica Bovina/química , Animales , Bovinos , Colesterol/administración & dosificación , Glucanos/administración & dosificación , Concentración de Iones de Hidrógeno , Nanogeles , Polietilenglicoles/administración & dosificación , Polietileneimina/administración & dosificación , Albúmina Sérica Bovina/administración & dosificaciónRESUMEN
Understanding the interface between microplastics and biological systems will provide new insights into the impacts of microplastics on living organisms. When microplastics enter the body, they are engulfed preferentially by phagocytes such as macrophages. However, it is not fully understood how phagocytes recognize microplastics and how microplastics impact phagocyte functions. In this study, we demonstrate that T cell immunoglobulin mucin 4 (Tim4), a macrophage receptor for phosphatidylserine (PtdSer) on apoptotic cells, binds polystyrene (PS) microparticles as well as multi-walled carbon nanotubes (MWCNTs) through the extracellular aromatic cluster, revealing a novel interface between microplastics and biological systems via aromatic-aromatic interactions. Genetic deletion of Tim4 demonstrated that Tim4 is involved in macrophage engulfment of PS microplastics as well as of MWCNTs. While Tim4-mediated engulfment of MWCNTs causes NLRP3-dependent IL-1ß secretion, that of PS microparticles does not. PS microparticles neither induce TNF-α, reactive oxygen species, nor nitric oxide production. These data indicate that PS microparticles are not inflammatory. The PtdSer-binding site of Tim4 contains an aromatic cluster that binds PS, and Tim4-mediated macrophage engulfment of apoptotic cells, a process called efferocytosis, was competitively blocked by PS microparticles. These data suggest that PS microplastics do not directly cause acute inflammation but perturb efferocytosis, raising concerns that chronic exposure to large amounts of PS microplastics may cause chronic inflammation leading to autoimmune diseases.
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Microplásticos , Nanotubos de Carbono , Humanos , Microplásticos/metabolismo , Plásticos/metabolismo , Poliestirenos/toxicidad , Poliestirenos/metabolismo , Mucina 4/metabolismo , Proteínas de la Membrana/genética , Macrófagos/metabolismo , Proteínas Portadoras , Apoptosis , InflamaciónRESUMEN
Multicellular tumor spheroids (MCTSs) are attractive for drug screening before animal tests because they emulate an in vivo microenvironment. The permeability of the MCTSs and tumor tissues towards the candidate drugs is not sufficient even though the drugs can penetrate monolayer cultured cells; therefore, nanocarriers are required to enhance permeability and deliver drugs. In this study, we prepared zwitterionic polymers of sulfobetaine methacrylates and (meth)acrylamides with or without hydroxy groups between the zwitterions to serve as highly permeable nanocarriers. In the sulfobetaine polymers, poly(2-hydroxy-3-((3-methacrylamidopropyl)dimethylammonio)propane-1-sulfonate), P(OH-MAAmSB), the hydroxy group containing methacrylamide polymer exhibited little cytotoxicity and membrane translocation ability against monolayer cultured cells. Moreover, the excellent permeability of the hepatocyte MCTS enabled P(OH-MAAmSB) to permeate it and reach the center region (â¼325 µm in diameter) at approximately 150 s, although poly(trimethyl-2-methacroyloxyethylammonium), a cationic polymer, penetrated just 1 to 2 layers from the periphery. The superior permeability of P(OH-MAAmSB) might be due to its good solubility and side chain conformation. P(OH-MAAmSB) is a promising nanocarrier with membrane translocation and permeability.
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Neoplasias , Polímeros , Animales , Betaína/análogos & derivados , Permeabilidad , Polímeros/química , Esferoides Celulares , Microambiente TumoralRESUMEN
Van der Waals (VDW) interactions provide fantastic properties for biological systems that function at room temperature. The VDW interaction, which primarily contributes to weak hydrogen bonding, is expected to play a key role in regulating hydrophobic hydration to express the biologically inert biocompatible function of polymerized MPCs (2-methacryloyloxyethyl phosphorylcholine). This report explores at the molecular level the biologically inert function of polymerized MPCs through an array of vibrational spectroscopic and computational characterization of MPC monomers, as temperature-dependent change of intramolecular weak hydrogen bonding. Synchrotron Fourier transform infrared microspectroscopy and terahertz time-domain spectroscopy were used to investigate temperature-dependent spectral changes in the low frequency vibrations of the MPC over the temperature range from cryogenic to room temperature, and the results were analysed by highly reliable well-established density functional theory (DFT) calculations. Complicated spectral features in the low frequency energy region and the uncertain conformations of the MPC in the amorphous powder state are clearly resolved under a polarizable continuum model and dispersion correction to pure DFT calculations.
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Polímeros , Enlace de Hidrógeno , Conformación Molecular , Interacciones Hidrofóbicas e HidrofílicasRESUMEN
This paper reports that water molecules around F-actin, a polymerized form of actin, are more mobile than those around G-actin or in bulk water. A measurement using pulse-field gradient spin-echo (1)H NMR showed that the self-diffusion coefficient of water in aqueous F-actin solution increased with actin concentration by â¼5%, whereas that in G-actin solution was close to that of pure water. This indicates that an F-actin/water interaction is responsible for the high self-diffusion of water. The local viscosity around actin was also investigated by fluorescence measurements of Cy3, a fluorescent dye, conjugated to Cys 374 of actin. The steady-state fluorescence anisotropy of Cy3 attached to F-actin was 0.270, which was lower than that for G-actin, 0.334. Taking into account the fluorescence lifetimes of the Cy3 bound to actin, their rotational correlation times were estimated to be 3.8 and 9.1ns for F- and G-actin, respectively. This indicates that Cy3 bound to F-actin rotates more freely than that bound to G-actin, and therefore the local water viscosity is lower around F-actin than around G-actin.
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Citoesqueleto de Actina/química , Agua/química , Animales , Tampones (Química) , Carbocianinas/química , Cisteína/química , Colorantes Fluorescentes/química , Resonancia Magnética Nuclear Biomolecular , Espectrometría de FluorescenciaRESUMEN
Tumor necrosis factor (TNF)-α exerts its biological function via TNF type 1 and type 2 receptors (TNFR1 and TNFR2). We have previously reported that bone resorption induced by lipopolysaccharide (LPS) in TNFR2-deficient mice is accelerated compared to that in wild-type (WT) mice. Although these results suggested that TNFR2 might have a protective role in bone resorption, we could not exclude the possibility that TNFR2 has no role in bone resorption. To clarify the role of TNFR2, we developed a TNF-α-induced bone resorption model using cholesterol-bearing pullulan nanogel as a TNF-α carrier to minimize the influence of inflammatory cytokines other than TNF-α. Injections of human TNF-α (hTNF), an agonist of mouse TNFR1, stimulated bone resorption lacunae on the calvariae in WT mice, but mouse TNF-α (mTNF), an agonist of both mouse TNFR1 and TNFR2, could not. To eliminate the possibility that the TNFR1 agonistic effects of hTNF were stronger than those of mTNF, we used the same model in TNFR2-deficient mice. Injection of mTNF resulted in clear bone resorption lacunae to the same extent observed after using hTNF in the TNFR2-deficient mice. Histomorphometric analysis of osteoclast number supported the observed changes in bone resorption lacunae. These data suggest that TNFR2 has a protective role in TNF-α-induced bone resorption.
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Resorción Ósea/inducido químicamente , Resorción Ósea/metabolismo , Receptores Tipo II del Factor de Necrosis Tumoral/metabolismo , Cráneo/efectos de los fármacos , Factor de Necrosis Tumoral alfa/farmacología , Animales , Densidad Ósea/efectos de los fármacos , Densidad Ósea/genética , Glucanos/química , Humanos , Ratones , Ratones Mutantes , Nanogeles , Polietilenglicoles/química , Polietileneimina/química , Receptores Tipo I de Factores de Necrosis Tumoral/agonistas , Receptores Tipo I de Factores de Necrosis Tumoral/metabolismo , Receptores Tipo II del Factor de Necrosis Tumoral/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Cráneo/citología , Cráneo/metabolismoRESUMEN
Cycloamylose (CA) exhibits differences in geometry and greater colloidal stability compared with amylose. Here we report the synthesis of a cationic CA derivative and its application for gene delivery. Cationic CA (catCA) and cationic amylose (catAmy) were synthesized by introducing spermine groups. The interactions between catCA or catAmy with plasmid DNA encoding firefly luciferase was examined by gel electrophoresis, dynamic light scattering, and transmission electron microscopy. Activity as a gene delivery system was evaluated by flow cytometry and luciferase assays. CatCA formed a condensed pDNA complex ( approximately 250 nm in size). The catCA complex showed enhanced cellular uptake and greater transfection efficiency than the catAmy complex. Hemolysis by membrane destabilization and the effects of hydroxychloroquine on transfection ability suggest that the formation of a supramolecular complex with CA is important for high transfection activity. These results suggested that CA can be used as new polysaccharide-based biomaterials.
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Materiales Biocompatibles , Ciclodextrinas/genética , Técnicas de Transferencia de Gen , Polisacáridos/genética , Animales , Materiales Biocompatibles/administración & dosificación , Materiales Biocompatibles/química , Células COS , Chlorocebus aethiops , Ciclodextrinas/administración & dosificación , Ciclodextrinas/química , Polisacáridos/administración & dosificación , Polisacáridos/química , Ovinos , Espermina/administración & dosificación , Espermina/químicaRESUMEN
Fast intracellular migration and controlled localization of molecules represent significant challenges for future applications of drug discovery and related fields. In this study, thermoresponsive sulfobetaine polymers with pyridinium cations are evaluated as biocompatible and mitochondria-localizing agents. Among the polymers, poly(3-(4-(2-methacrylamido)ethyl pyridinio-1-yl)propane-1-sulfonate), P(E-PySMAAm)14k (Mn = 14 000 g mol-1 ) exhibit thermoresponsiveness with an upper critical solution temperature like behavior in cell culture medium containing serum with minimal cytotoxicity. Upon the addition of P(E-PySMAAm)14k to HeLa cells at temperatures above the clearing point at 37 °C, effective localization is observed in mitochondria. However, increased intensity but nonspecific localization is observed below the clearing point at 4 °C. Doxorubicin is conjugated to the P(E-PySMAAm) and achieves effective mitochondrial delivery while maintaining drug efficacy. Such sulfobetaine polymers represent promising tools for intracellular delivery of molecules.
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Betaína/análogos & derivados , Sistemas de Liberación de Medicamentos , Mitocondrias/efectos de los fármacos , Nanopartículas/química , Betaína/química , Betaína/farmacología , Cationes/química , Cationes/farmacología , Movimiento Celular/efectos de los fármacos , Doxorrubicina/química , Doxorrubicina/farmacología , Células HeLa , Humanos , Mitocondrias/patología , Polímeros/química , Polímeros/farmacología , TemperaturaRESUMEN
A flexible biosensor using a phospholipid polymer to immobilization of glucose oxidase (GOD) was fabricated and tested. At first, an enzyme membrane formed by immobilizing GOD onto a porous polytetrafluoroethylene (PTFE) membrane using the phospholipid polymer (2-methacryloyloxyethyl phosphorylcholine (MPC) copolymerized with 2-ethylhexylmethacrylate (EHMA):PMEH) was evaluated. According to the result of amperometric measurement, average density of GOD to be immobilized was optimized to 38.9 units cm(-2). Temperature and pH dependences were also investigated. Then, a flexible glucose sensor was fabricated by immobilizing GOD onto a flexible hydrogen peroxide electrode using PMEH. The flexible glucose sensor showed a linear relationship between output currents and glucose concentration in 0.05-1.00 mmol L(-1), with a correlation coefficient of 0.999. The calibration range covered the normal tear glucose level of 0.14-0.23 mmol L(-1). This indicates that the flexible biosensor is considered to be useful for monitoring of glucose in tear fluids.
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Técnicas Biosensibles/instrumentación , Técnicas Biosensibles/métodos , Glucosa/análisis , Membranas Artificiales , Metacrilatos/química , Fosforilcolina/análogos & derivados , Politetrafluoroetileno/química , Enzimas Inmovilizadas/química , Glucosa Oxidasa/química , Humanos , Fosforilcolina/química , Docilidad , Sensibilidad y EspecificidadRESUMEN
Assessment of van der Waals (VDW) interactions is fundamental to all of the central quest of structure that regulates the biological function. VDW interactions contributing to intramolecular weak hydrogen bonding are regarded as an important force to regulate the thermal stimuli-sensitive function of sulfobetaine methacrylate, DMAPS. We present here the conversion from the thermal-motion form at room temperature to the weak-hydrogen-bonded form against thermal motion as a terahertz spectral change with a definite isosbestic point from an absorption peak of one form to the other. Vibrational absorptions are used as a probe for assessing VDW interactions in conjunction with highly reliable and well-established density functional theory (DFT) calculations for analysis. Complicated spectral features and uncertain conformations of DMAPS in the amorphous state are clearly resolved under the polarizable continuum model and the dispersion correction for the pure DFT calculations.