RESUMO
Thorough characterization of toxic effects of nanoparticles (NP) is desirable due to the increasing risk of potential environmental contamination by NP. In the current study, we combined three recently developed proteomics approaches to assess the effects of Au, CuO, and CdTe NP on the innate immune system. The human monocyte cell line THP-1 was employed as a model. The anticancer drugs camptothecin and doxorubicin were used as positive controls for cell death, and lipopolysaccharide was chosen as a positive control for proinflammatory activation. Despite equivalent overall toxicity effect (50 ± 10% dead cells), the three NP induced distinctly different proteomics signatures, with the strongest effect being induced by CdTe NP, followed by CuO and gold NP. The CdTe toxicity mechanism involves down-regulation of topoisomerases. The effect of CuO NP is most reminiscent of oxidative stress and involves up-regulation of proteins involved in heat response. The gold NP induced up-regulation of the inflammatory mediator, NF-κB, and its inhibitor TIPE2 was identified as a direct target of gold NP. Furthermore, gold NP triggered activation of NF-κB as evidenced by phosphorylation of the p65 subunit. Overall, the combined proteomics approach described here can be used to characterize the effects of NP on immune cells.
Assuntos
Imunidade Inata/efeitos dos fármacos , Inflamação/genética , Nanopartículas Metálicas/efeitos adversos , Proteoma/genética , Proteômica , Compostos de Cádmio/efeitos adversos , Camptotecina/administração & dosagem , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cobre/efeitos adversos , Citotoxinas/efeitos adversos , Doxorrubicina/administração & dosagem , Ouro/efeitos adversos , Humanos , Imunidade Inata/genética , Inflamação/induzido quimicamente , Lipopolissacarídeos/administração & dosagem , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Proteoma/efeitos dos fármacos , Telúrio/efeitos adversosRESUMO
The surface chemistry of nanoparticles (NPs) is one of the critical factors determining their cellular responses. In this study, the cytotoxicity and genotoxicity of copper oxide (CuO) NPs with a similar size but different surface chemistry to rat bone marrow mesenchymal stem cells (MSCs) were investigated. The morphology, size and surface charge of four types of CuO NPs, i.e., CuO-core, CuO-COOH, CuO-NH2 and CuO-PEG NPs, were characterized by TEM, dynamic light scattering (DLS) and zeta-potential measurement, respectively. All of the four CuO NPs had a negative surface charge around -10 mV and showed a similar tendency to form agglomerates with a size of -200 nm in cell culture environment. The cytotoxicity of CuO NPs to MSCs at various concentrations and incubation periods were firstly evaluated. The CuO NPs showed dose-dependent and time-dependent toxicity to MSCs, and their surface chemistry had influence on the toxicity to some extent too. The intracellular reactive oxygen species (ROS) level of MSCs was then quantified. Finally, the genotoxicity of the CuO NPs was studied by comet assay. The results suggest that the genotoxicity of CuO NPs was mainly dependent on NPs concentration, and was only slightly influenced by their surface chemistry. The osteogenic and adipogenic differentiation abilities of the MSCs exposed to different CuO NPs were studied by Alizarin Res S and Oil Red O staining. The preliminary results showed that the exposure to 10 µg/mL CuO NPs will, not lead to significant impact on the differentiation potential of the MSCs.
Assuntos
Cobre/química , Cobre/toxicidade , Células-Tronco Mesenquimais/efeitos dos fármacos , Mutagênicos/química , Mutagênicos/toxicidade , Nanopartículas/toxicidade , Animais , Diferenciação Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Espaço Intracelular/efeitos dos fármacos , Espaço Intracelular/metabolismo , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Propriedades de Superfície , Fatores de TempoRESUMO
Drug delivery monitoring and tracking in the human body are two of the biggest challenges in targeted therapy to be addressed by nanomedicine. The ability of imaging drugs and micro-/nanoengineered drug carriers and of visualizing their interactions at the cellular interface in a label-free manner is crucial in providing the ability of tracking their cellular pathways and will help understand their biological impact, allowing thus to improve the therapeutic efficacy. We present a fast, label-free technique to achieve high-resolution imaging at the mid-infrared (MIR) spectrum that provides chemical information. Using our custom-made benchtop infrared microscope using a high-repetition-rate pulsed laser (80 MHz, 40 ps), we were able to acquire images with subwavelength resolution (0.8 × λ) at very high speeds. As a proof-of-concept, we embarked on the investigation of nanoengineered polyelectrolyte capsules (NPCs) containing the anticancer drug, docetaxel. These NPCs were synthesized using a layer-by-layer approach built upon a calcium carbonate (CaCO3) core, which was then removed away with ethylenediaminetetraacetic acid. The obtained MIR images show that NPCs are attached to the cell membrane, which is a good step toward an efficient drug delivery. This has been confirmed by both three-dimensional confocal fluorescence and stimulated emission depletion microscopy. Coupled with additional instrumentation and data processing advancements, this setup is capable of video-rate imaging speeds and will be significantly complementing current super-resolution microscopy techniques while providing an unperturbed view into living cells.
RESUMO
We propose and demonstrate a novel subtraction microscopy algorithm, exploiting fluorescence emission difference or switching laser mode and their derivatives for image enhancement. The key novelty of the proposed approach lies in the weighted subtraction coefficient, adjusted pixel-by-pixel with respect to the intensity distributions of initial images. This method produces significant resolution enhancement and minimizes image distortions. Our theoretical and experimental studies demonstrate that this approach can be applied to any optical microscopy techniques, including label free and non-linear methods, where common super-resolution techniques cannot be used.
RESUMO
We present the concept of multifunctional nanoengineered polymer capsules and outline their applications as new drug delivery systems or supramolecular toolboxes containing, for example, enzymes capable of converting nontoxic prodrugs into toxic drugs at a designated location. Such functionalized nanocontainers offer a wide range of applications including enzymatic catalysis, controlled release, and directed drug delivery in medicine due to their multifunctionality. The unique advantage of capsules in comparison to other systems is that they can be functionalized or loaded simultaneously with the above-mentioned components, thus permitting multifunctional processes in single cells.
Assuntos
Sistemas de Liberação de Medicamentos , Nanocápsulas/administração & dosagem , Nanocápsulas/química , Nanopartículas/química , Polímeros/química , Animais , Neoplasias da Mama/terapia , Cápsulas , Catálise , Enzimas/química , Corantes Fluorescentes/farmacologia , Humanos , Magnetismo , Microscopia de Fluorescência , Nanotecnologia/métodos , Pontos QuânticosRESUMO
This review is devoted to a novel type of polymer micro- and nanocapsules. The shell of the capsule is fabricated by alternate adsorption of oppositely charged polyelectrolytes (PEs) onto the surface of colloidal particles. Cores of different nature (organic or inorganic) with size varied from 0.1 to 10 mum can be used for templating such PE capsules. The shell thickness can be tuned in nanometer range by assembling of defined number of PE layers. The permeability of capsules depends on the pH, ionic strength, solvent, polymer composition, and shell thickness; it can be controlled and varied over wide range of substances regarding their molecular weight and charge. Including functional polymers into capsule wall, such as weak PEs or thermosensitive polymers, makes the capsule permeability sensitive to correspondent external stimuli. Permeability of the capsules is of essential interest in diverse areas related to exploitation of systems with controlled and sustained release properties. The envisaged applications of such capsules/vesicles cover biotechnology, medicine, catalysis, food industry, etc.
Assuntos
Cápsulas/química , Adsorção , Materiais Biocompatíveis/química , Fenômenos Químicos , Físico-Química , Concentração de Íons de Hidrogênio , Peso Molecular , Tamanho da Partícula , Permeabilidade , Solventes/química , Propriedades de SuperfícieRESUMO
Cell-sized polyelectrolyte capsules functionalized with a redox-driven proton pump protein were assembled for the first time. The interaction of polyelectrolyte microcapsules, fabricated by electrostatic layer-by-layer assembly, with cytochrome c oxidase molecules was investigated. We found that the cytochrome c oxidase retained its functionality, that the functionalized microcapsules interacting with cytochrome c oxidase were permeable and that the permeability characteristics of the microcapsule shell depend on the shell components. This work provides a significant input towards the fabrication of an integrated device made of biological components and based on specific biomolecular functions and properties.
Assuntos
Proteínas de Bactérias/química , Complexo IV da Cadeia de Transporte de Elétrons/química , Enzimas Imobilizadas/química , Thermus thermophilus/enzimologia , CápsulasAssuntos
Cápsulas/química , Cápsulas/efeitos da radiação , Preparações de Ação Retardada/química , Preparações de Ação Retardada/efeitos da radiação , Portadores de Fármacos/química , Portadores de Fármacos/efeitos da radiação , Sonicação , Materiais Revestidos Biocompatíveis/química , Materiais Revestidos Biocompatíveis/efeitos da radiação , Difusão , Teste de Materiais , Nanoestruturas/química , Nanoestruturas/ultraestrutura , Nanotecnologia/métodos , Tamanho da Partícula , Preparações Farmacêuticas/administração & dosagem , Preparações Farmacêuticas/químicaRESUMO
We show that polyelectrolyte shells may be perfect hosts and microreactors for catalysis. Using a layer-by-layer self-assembly process to form hollow polyelectrolyte capsules, we show that individual layers can be replaced by palladium and gold nanoclusters forming robust cell-type microcapsules. Testing the catalytic activity of those embedded nanoclusters using the Sonogashira cross-coupling reaction, we show that simple self-assembled polyelectrolyte shells are ideal as carrier of at least one expensive catalyst, and possibly several, opening the road to new cascade reactions.
Assuntos
Nanocápsulas/química , Paládio/química , Catálise , Membranas Artificiais , Nanocápsulas/ultraestrutura , Polímeros/químicaRESUMO
The objective of the present investigation was to fabricate composite colloidal particles consisting of a sacrificial, decomposable template of biodegradable nature covered with biocompatible polyelectrolyte multilayers using the layer-by-layer sequential adsorption technique. Poly-dl-lactic acid and poly(dl-lactic-co-glycolic acid) were chosen to design the microparticulate template, and a preliminary feasibility study was carried out with poly(styrene sulfonate sodium)-poly(allylamine hydrochloride) as shell components. The properties of both core-shell and hollow structures obtained by core dissolution were characterized by confocal laser scanning microscopy, microelectrophoresis, scanning force microscopy, and scanning electron microscopy. The concept was then extended to biocompatible polyelectrolytes as shell wall building blocks to deduce stable hollow capsules with tailored properties. Uniform, complete coating with oppositely charged polyelectrolyte pairs was achieved for all the combinations investigated. The results demonstrate that polyester microparticles could serve as viable alternative components to conventionally employed templates to derive hollow capsules with defined size, shape, and shell thickness. With all the components used for fabrication being biocompatible, these polyelectrolyte capsules may find interesting applications in the fields of biology, biochemistry, biotechnology, and drug delivery.
Assuntos
Materiais Biocompatíveis , Microscopia Eletrônica de Varredura , Microesferas , Estrutura MolecularRESUMO
We present a novel method for remote release of an encapsulated material from polyelectrolyte capsules based on laser light illumination. Two different components were introduced in the polyelectrolyte shells of PAH/PSS capsules-either Ag nanoparticles or IR dye-to induce absorption of light. Under laser illumination the capsules containing Ag nanoparticles or IR dye were deformed or cut, thus providing a venue for remote release of encapsulated materials. The experiments were conducted with a low-power near-infrared continuous-wave laser diode.
RESUMO
A novel, moderately thermophilic, strictly anaerobic, mixotrophic bacterium, designated strain LF13T, was isolated from a deep-sea hydrothermal chimney sample that was collected at a vent site at 14 degrees 45' N, 44 degrees 59' W on the Mid-Atlantic Ridge. Cells were Gram-negative, thin, non-motile rods of variable length. Strain LF13T grew optimally at pH 6.8-7.0 and 60 degrees C with 2.5% (w/v) NaCl. It grew chemo-organoheterotrophically, fermenting proteinaceous substrates, pyruvate and Casamino acids. The strain was able to grow by respiration, utilizing molecular hydrogen (chemolithoheterotrophically) or acetate as electron donors and nitrate as an electron acceptor. Ammonium was formed in the course of denitrification. One-hundred milligrams of yeast extract per litre were required for growth of the strain. The G + C content of the genomic DNA of strain LF13T was 42.5 mol%. Neither 16S rDNA sequence similarity values nor phylogenetic analysis unambiguously related strain LF13T with members of any recognized bacterial phyla. On the basis of 16S rDNA sequence comparisons, and in combination with physiological and morphological traits, a novel genus, Caldithrix, is proposed, with strain LF13T (= DSM 13497T =VKM B-2286T) representing the type species, Caldithrix abyssi.