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Cellular delivery of plasmid DNA (pDNA) specifically into dendritic cells (DCs) has provoked wide attention in various applications. However, delivery tools that achieve effective pDNA transfection in DCs are rare. Herein, we report that tetrasulphide bridged mesoporous organosilica nanoparticles (MONs) have enhanced pDNA transfection performance in DC cell lines compared to conventional mesoporous silica nanoparticles (MSNs). The mechanism of enhanced pDNA delivery efficacy is attributed to the glutathione (GSH) depletion capability of MONs. Reduction of initially high GSH levels in DCs further increases the mammalian target of rapamycin complex 1 (mTORc1) pathway activation, enhancing translation and protein expression. The mechanism was further validated by showing that the increased transfection efficiency was apparent in high GSH cell lines but not in low GSH ones. Our findings may provide a new design principle of nano delivery systems where the pDNA delivery to DCs is important.
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Rapid and continuous ammonium adsorption from mainstream coupled with side-stream ammonium recovery and adsorbent regeneration could enable ammonium recovery from domestic wastewater. This study describes the use of tailored poly(acrylic acid)-based (NaPAA) hydrogels as effective sorbents for ammonium removal from domestic wastewater. Modified NaPAA hydrogels having 60% ionization and 4.8 mol % N',N'-methylenebisacrylamide as the cross-linker reduced the overall swelling by 92% from 407 to 31 g/g because of higher cross-linking density. At hydrogel loadings of 2.5-7.5 g/L, the NaPAA hydrogels achieved ammonium concentrations of 8.3 ± 0.6 to 10.1 ± 0.1 mg/L NH4-N, which corresponds to removal efficiencies of 53-77% after 10 min of contact time in real domestic wastewater. At the same hydrogel loadings, the ammonium removal efficiency of NaPAA hydrogels in synthetic wastewater was found to be comparable to that in real sewage (71% vs 69%, respectively), suggesting that the sorption performance is only marginally affected by organic constituents found in domestic wastewater. In addition, the NaPAA hydrogels removed 25-51% ammonium in 10 min from synthetic streams having 200-400% higher ionic strengths than those commonly observed in sewage. Furthermore, simulation studies showed that a discharge concentration of â¼1.9 mg/L NH4-N, well below the commonly applied discharge limits in most regions, can be achieved using mainstream ammonium removal by NaPAA hydrogels followed by biological assimilation from the growth of ordinary heterotrophic organisms.
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Compuestos de Amonio , Aguas Residuales , Resinas Acrílicas , HidrogelesRESUMEN
Type 2 diabetes makes up approximately 85% of all diabetic cases and it is linked to approximately one-third of all hospitalisations. Newer therapies with long-acting biologics such as glucagon-like peptide-1 (GLP-1) analogues have been promising in managing the disease, but they cannot reverse the pathology of the disease. Additionally, their parenteral administration is often associated with high healthcare costs, risk of infections, and poor patient adherence associated with phobia of needles. Oral delivery of these compounds would significantly improve patient compliance; however, poor enzymatic stability and low permeability across the gastrointestinal tract makes this task challenging. In the present work, large pore dendritic silica nanoparticles (DSNPs) with a pore size of ~10 nm were prepared, functionalized, and optimized in order to achieve high peptide loading and improve intestinal permeation of exenatide, a GLP-1 analogue. Compared to the loading capacity of the most popular, Mobil Composition of Matter No. 41 (MCM-41) with small pores, DSNPs showed significantly high loading owing to their large and dendritic pore structure. Among the tested DSNPs, pristine and phosphonate-modified DSNPs (PDSNPs) displayed remarkable loading of 40 and 35% w/w, respectively. Furthermore, particles successfully coated with positively charged chitosan reduced the burst release of exenatide at both pH 1.2 and 6.8. Compared with free exenatide, both chitosan-coated and uncoated PDSNPs enhanced exenatide transport through the Caco-2 monolayer by 1.7 fold. Interestingly, when a triple co-culture model of intestinal permeation was used, chitosan-coated PDSNPs performed better compared to both PDSNPs and free exenatide, which corroborated our hypothesis behind using chitosan to interact with mucus and improve permeation. These results indicate the emerging role of large pore silica nanoparticles as promising platforms for oral delivery of biologics such as exenatide.
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Mesoporous silica nanoparticles (MSNs) are drug delivery agents that are able to incorporate drugs within their pores. Furthermore, MSNs can be functionalized by attachment of bioactive ligands on their surface to enhance their activity, and nanoparticles modified with glycosaminoglycan (GAG) mimetics inhibit the entry of herpes simplex virus (HSV) into cells. In this study, structure-activity relationships of GAGs attached to MSNs were investigated in relation to HSV-1 and HSV-2, and acyclovir was loaded into the pores of MSNs. The sulfonate group was demonstrated to be essential for antiviral activity, which was enhanced by incorporating a benzene group within the ligand. Loading acyclovir into GAG mimetic-functionalized MSNs reduced the viral infection, resulting in nanoparticles that simultaneously target two distinct viral pathways, namely, inhibition of viral entry and inhibition of DNA replication.
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Conventional oral drug formulations for colonic diseases require the administration of high doses of drug to achieve effective drug concentrations at the target site. However, this exposes patients to serious systemic toxicity in order to achieve efficacy. To overcome this problem, an oral drug delivery system was developed by loading a large amount (ca. 34% w/w) of prednisolone into 3-aminopropyl-functionalized mesoporous silica nanoparticles (MCM-NH2) and targeting prednisolone release to the colon by coating the nanoparticle with succinylated ε-polylysine (SPL). We demonstrate for the first time the pH-responsive ability of SPL as a "nanogate" to selectively release prednisolone in the pH conditions of the colon (pH 5.5-7.4) but not in the more acidic conditions of the stomach (pH 1.9) or small intestine (pH 5.0). In addition to targeting drug delivery to the colon, we explored whether the nanoparticles could deliver cargo intracellularly to immune cells (RAW 264.7 macrophages) and intestinal epithelial cells (LS 174T and Caco-2 adenocarcinoma cell lines). To trace uptake, MCM-NH2 were loaded with a cell membrane-impermeable dye, sulforhodamine B. The SPL-coated nanoparticles were able to deliver the dye intracellularly to RAW 264.7 macrophages and the intestinal epithelial cancer cells, which offers a highly promising and novel drug delivery system for diseases of the colon such as inflammatory bowel disease and colorectal cancer.
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Nanopartículas , Animales , Células CACO-2 , Sistemas de Liberación de Medicamentos , Humanos , Concentración de Iones de Hidrógeno , Ratones , Polilisina , Porosidad , Dióxido de SilicioRESUMEN
A glycosaminoglycan mimetic was attached to the surface of solid and mesoporous silica nanoparticles to create novel antiviral agents against herpes simplex type 1 and type 2 viruses. The nanoparticles act as viral entry inhibitors that appear to block viral attachment and penetration into susceptible cells.
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Antivirales/farmacología , Herpesvirus Humano 1/efectos de los fármacos , Herpesvirus Humano 2/efectos de los fármacos , Nanopartículas , Internalización del Virus/efectos de los fármacos , Animales , Bencenosulfonatos , Chlorocebus aethiops , Glicosaminoglicanos , Herpes Simple , Dióxido de Silicio , Células Vero , Ensayo de Placa ViralRESUMEN
We introduce "sense, track and separate" approach for the removal of Hg(2+) ion from aqueous media using highly ordered and magnetic mesoporous ferrosilicate nanocages functionalised with rhodamine fluorophore derivative. These functionalised materials offer both fluorescent and magnetic properties in a single system which help not only to selectively sense the Hg(2+) ions with a high precision but also adsorb and separate a significant amount of Hg(2+) ion in aqueous media. We demonstrate that the magnetic affinity of these materials, generated from the ultrafine γ-Fe2O3 nanoparticles present inside the nanochannels of the support, can efficiently be used as a fluorescent tag to sense the Hg(2+) ions present in NIH3T3 fibroblasts live cells and to track the movement of the cells by external magnetic field monitored using confocal fluorescence microscopy. This simple approach of introducing multiple functions in the magnetic mesoporous materials raise the prospect of creating new advanced functional materials by fusing organic, inorganic and biomolecules to create advanced hybrid nanoporous materials which have a potential use not only for sensing and the separation of toxic metal ions but also for cell tracking in bio-separation and the drug delivery.
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Mercurio/análisis , Microscopía Fluorescente , Nanoestructuras/química , Espectrometría de Fluorescencia , Animales , Colorantes Fluorescentes/química , Iones/química , Nanopartículas de Magnetita/química , Metales/química , Ratones , Células 3T3 NIH , Porosidad , Rodaminas/química , Silanos/química , Difracción de Rayos XRESUMEN
This paper reports the plasticisation effect of the ionic liquid, 1-ethyl-3-methylimidazolium acetate ([Emim][OAc]), as compared with the traditionally used plasticiser, glycerol, on the characteristics of starch-based films. For minimising the additional effect of processing, a simple compression moulding process (which involves minimal shear) was used for preparation of starch-based films. The results show that [Emim][OAc] was favourable for plasticisation, i.e., disruption of starch granules (by scanning electron microscopy), and could result in a more amorphous structure in the starch-based materials (by X-ray diffraction and dynamic mechanical analysis). (13)C CP/MAS and SPE/MAS NMR spectroscopy revealed that not only was the crystallinity reduced by [Emim][OAc], but also the amorphous starch present was plasticised to a more mobile form as indicated by the appearance of amorphous starch in the SPE/MAS spectrum. Mechanical results illustrate that, when either glycerol or [Emim][OAc] was used, a higher plasticiser content could contribute to higher flexibility. In spite of the accelerated thermal degradation of starch by [Emim][OAc] as shown by thermogravimetric analysis, the biodegradation study revealed the antimicrobial effect of [Emim][OAc] on the starch-based materials. Considering the high-amylose starch used here which is typically difficult to gelatinise in a traditional plasticiser (water and/or glycerol), [Emim][OAc] is demonstrated to be a promising plasticiser for starch to develop "green" flexible antimicrobial materials for novel applications.
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Acetatos/química , Glicerol/química , Imidazoles/química , Plastificantes/química , Almidón/química , Cristalización , Módulo de Elasticidad , Líquidos Iónicos/químicaRESUMEN
MOFs-based mixed matrix membranes (MMMs) have attracted extensive attention in recent years due to their potential high separation performance, the low cost, and good mechanical properties. However, it is still very challenging to achieve defect-free interface between micrometer-sized MOFs and a polymer matrix. In this study, [Cd2L(H2O)]2·5H2O (Cd-6F) synthesized using 4,4'-(hexafluoroisopropylidene)diphthalic anhydride (6FDA) as an organic ligand was introduced into the 6FDA-ODA polyimide matrix to achieve novel MOF MMMs. A specific interfacial interaction between MOF crystals and polymer chains was innovatively targeted and achieved through in situ polymerization procedure. The enhanced adhesion between MOF particles and polymer phase was observed, and the improved interfacial interaction between Cd-6F and the 6FDA-ODA polyimide matrix was confirmed by detailed characterizations including FTIR and NMR. In the meantime, the gas permeance and selectivity of the MMMs are strongly dependent on their morphology. The MMM derived from in situ polymerization presents excellent interfaces between micrometer-sized MOF crystals and the polymer matrix, resulting in increased permeability and selectivity. The strategy shown here can be further utilized to select the MOF/polymer pair, eliminate interfacial voids, and improve membrane separation performance of MOFs-based MMMs.
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The development of functional nanocarriers that can enhance the cellular delivery of a variety of nucleic acid agents is important in many biomedical applications such as siRNA therapy. We report the synthesis of large pore mesoporous silica nanoparticles (LPMSN) loaded with iron oxide and covalently modified by polyethyleneimine (denoted PEI-Fe-LPMSN) as carriers for gene delivery. The LPMSN have a particle size of â¼200 nm and a large pore size of 11 nm. The large pore size is essential for the formation of large iron oxide nanoparticles to increase the magnetic properties and the adsorption capacity of siRNA molecules. The magnetic property facilitates the cellular uptake of nanocarriers under an external magnetic field. PEI is covalently grafted on the silica surface to enhance the nanocarriers' affinity against siRNA molecules and to improve gene silencing performance. The PEI-Fe-LPMSN delivered siRNA-PLK1 effectively into osteosarcoma cancer cells, leading to cell viability inhibition of 80%, higher compared to the 50% reduction when the same dose of siRNA was delivered by a commercial product, oligofectamine.
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Técnicas de Transferencia de Gen , Nanopartículas de Magnetita/química , Dióxido de Silicio/química , Adsorción , Transporte Biológico Activo , Proteínas de Ciclo Celular/antagonistas & inhibidores , Proteínas de Ciclo Celular/genética , Línea Celular Tumoral , Terapia Genética/métodos , Humanos , Nanopartículas de Magnetita/administración & dosificación , Nanopartículas de Magnetita/ultraestructura , Nanotecnología , Oligodesoxirribonucleótidos/administración & dosificación , Oligodesoxirribonucleótidos/farmacocinética , Osteosarcoma/enzimología , Osteosarcoma/genética , Osteosarcoma/terapia , Polietileneimina/química , Porosidad , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , Proteínas Serina-Treonina Quinasas/genética , Proteínas Proto-Oncogénicas/antagonistas & inhibidores , Proteínas Proto-Oncogénicas/genética , ARN Interferente Pequeño/administración & dosificación , ARN Interferente Pequeño/farmacocinética , Quinasa Tipo Polo 1RESUMEN
A novel water-soluble macromonomer based on octavinyl silsesquioxane has been synthesized and contains vinyl-terminated PEG 400 in each of the eight arms to promote water solubility. The macromonomer was characterized by NMR and FTIR and its aqueous solution properties examined. In water it exhibits an LCST with a cloud point at 23 °C for a 10 wt % aqueous solution. It is surface active with a CMC of 1.5 × 10(-5) M in water and in 20:80 v/v acetone/water the CMC is 7.1 × 10(-5) M, and TEM images showed spherical 22 nm aggregates in aqueous solution above the CMC. The macromonomer was copolymerized in a 20:80 v/v acetone/water mixture with a vinyl-terminated, triblock copolymer of lactide-PEG-lactide to form a library of cross-linked hydrogels that were designed for use as scaffolds for alveolar bone repair. The cross-linked copolymer networks were shown to contain a range of nm-µm sized pores and their swelling properties in water and PBS at pH 7.4 were examined. At pH 7.4 the hydrogel networks undergo a slow hydrolysis with the release of principally PEG and lactic acid fragments. The hydrogels were shown to be noncytotoxic toward fibroblast cultures at pH 7.4, both initially (days 1-5) and after significant hydrolysis had taken place (days 23-28).
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Fibroblastos/química , Hidrogeles/síntesis química , Compuestos de Organosilicio/química , Polietilenglicoles/química , Animales , Células Cultivadas , Femenino , Fibroblastos/citología , Hidrogeles/química , Sustancias Macromoleculares/síntesis química , Sustancias Macromoleculares/química , Tamaño de la Partícula , Ratas , Ratas Endogámicas Lew , Propiedades de SuperficieRESUMEN
Understanding the nature of mixed surfactant self-assembly on the surface of organoclays is an important step toward optimizing their performance in polymer nanocomposites and for other potential applications, where selective surface interactions are crucial. In segmented thermoplastic polyurethane nanocomposite systems, dual-modified organoclays have shown significantly better performance compared to their single-modified counterparts. Until now, we had not fully characterized the physical chemistry of these dual-modified layered silicates, but had hypothesized that the enhanced composite performance arises due to some degree of nanoscale phase separation on the nanofiller surface, which enables enhanced compatibilization and more specific and inclusive interactions with the nanoscale hard and soft domains in these thermoplastic elastomers. This work examines the organization of quaternary alkyl ammonium compounds on the surface of Lucentite SWN using X-ray diffraction (XRD), thermogravimetric analysis (TGA), attenuated total reflectance Fourier-transfer infrared (ATR FT-IR), (13)C cross-polarization (CP)/magic angle spinning (MAS) nuclear magnetic resonance (NMR), and small-angle neutron scattering (SANS). When used in combination with choline, dimethyldioctadecylammonium (DMDO) was observed to self-assemble into discontinuous hydrophobic domains. The inner part of these hydrophobic domains was essentially unaffected by the choline (CC); however, surfactant intermixing was observed either at the periphery or throughout the choline-rich phase surrounding those domains.
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Colina/química , Compuestos de Amonio Cuaternario/química , Silicatos/síntesis química , Silicatos/química , Propiedades de SuperficieRESUMEN
The effects of moisture and thermal denaturation on the solid-state structure and molecular mobility of soy glycinin powder were investigated using multiple techniques that probe over a range of length and time scales. In native glycinin, increased moisture resulted in a decrease in both the glass transition temperature and the denaturation temperature. The sensitivity of the glass transition temperature to moisture is shown to follow the Gordon-Taylor equation, while the sensitivity of the denaturation temperature to moisture is modeled using Flory's melting point depression theory. While denaturation resulted in a loss of long-range order, the principal conformational structures as detected by infrared are maintained. The temperature range over which the glass to rubber transition occurred was extended on the high temperature side, leading to an increase in the midpoint glass transition temperature and suggesting that the amorphous regions of the newly disordered protein are less mobile. (13)C NMR results supported this hypothesis.
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Globulinas/química , Glycine max/química , Proteínas de Soja/química , Rastreo Diferencial de Calorimetría , Vidrio/química , Globulinas/metabolismo , Humedad , Espectroscopía de Resonancia Magnética , Movimiento (Física) , Conformación Proteica , Desnaturalización Proteica , Soluciones , Proteínas de Soja/metabolismo , Espectroscopía Infrarroja por Transformada de Fourier , Termodinámica , Temperatura de TransiciónRESUMEN
The movement of water into harvest-ripe grains of dormant and non-dormant genotypes of wheat (Triticum aestivum L.) was investigated using Magnetic Resonance Micro-Imaging (MRMI). Images of virtual sections, both longitudinal and transverse, throughout the grain were collected at intervals after the start of imbibition and used to reconstruct a picture of water location within the different grain tissues and changes over time. The observations were supplemented by the weighing measurements of water content and imbibition of grains in water containing I(2)/KI which stains starch and lipid, thereby acting as a marker for water. In closely related genotypes, with either a dormant or a non-dormant phenotype, neither the rate of increase in water content nor the pattern of water distribution within the grain was significantly different until 18 h, when germination became apparent in the non-dormant genotype. Water entered the embryo and scutellum during the very early stages of imbibition through the micropyle and by 2 h water was clearly evident in the micropyle channel. After 12 h of imbibition, embryo structures such as the coleoptile and radicle were clearly distinguished. Although water accumulated between the inner (seed coat) and outer (pericarp) layers of the coat surrounding the grain, there was no evidence for movement of water directly across the coat and into the underlying starchy endosperm.
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Triticum/fisiología , Agua/metabolismo , Grano Comestible/química , Grano Comestible/genética , Grano Comestible/fisiología , Germinación , Imagen por Resonancia Magnética , Semillas/química , Semillas/genética , Semillas/fisiología , Triticum/química , Triticum/genéticaRESUMEN
We report a multitechnique study of structural organization and molecular mobility for soy glycinin at a low moisture content (<30% w/w) and relate these to its glass-to-rubber transition. Small-angle X-ray scattering (SAXS), differential scanning calorimetry (DSC), Fourier transform infrared (FTIR) spectroscopy, and nuclear magnetic resonance (NMR) spectroscopy are used to probe structure and mobility on different length and time scales. NMR (approximately 10(-6) to 10(-3) s) reveals transitions at a higher moisture content (>17%) than DSC or SAXS, which sample for much longer times (approximately 10 to 10(3) s) and where changes are detected at >13% water content at 20 degrees C. The mobility transitions are accompanied by small changes in unit-cell parameters and IR band intensities and are associated with the enhanced motion of the polypeptide backbone. This study shows how characteristic features of the ordered regions of the protein (probed by SAXS and FTIR) and mobile segments (probed by NMR and DSC) can be separately monitored and integrated within a mobility transformation framework.
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Globulinas/química , Glycine max/metabolismo , Proteínas de Soja/química , Rastreo Diferencial de Calorimetría/métodos , Concentración de Iones de Hidrógeno , Espectroscopía de Resonancia Magnética , Conformación Molecular , Conformación Proteica , Estructura Secundaria de Proteína , Proteínas/química , Dispersión de Radiación , Espectrofotometría Infrarroja/métodos , Espectroscopía Infrarroja por Transformada de Fourier , Temperatura , Agua/química , Rayos XRESUMEN
The effect of moisture content and temperature on water diffusion into a modified high amylose (< or = 90%) maize thermoplastic starch blend was investigated. Gravimetric and magnetic resonance imaging (MRI) studies were conducted to elucidate the diffusion mechanism and diffusion coefficients for this system. The diffusion coefficient data demonstrated that the rate of water diffusion into this blend was significantly dependent upon temperature and moisture content. Water diffusion was faster at higher temperatures and generally for samples stored at higher relative humidity environments. It was revealed from the gravimetric data that water diffusion into this starch blend was Fickian; however, further analysis of the MRI images found that the water diffusion mechanism was exponentially dependent on the concentration. This result was determined by comparing experimental water concentration profiles to a theoretical model calculated using the implicit Crank-Nicolson finite difference method.