Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 10 de 10
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
Beilstein J Nanotechnol ; 5: 2479-88, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25671143

RESUMO

In this work, we study epithelial cell growth on substrates decorated with gold nanorods that are functionalized either with a positively charged cytotoxic surfactant or with a biocompatible polymer exhibiting one of two different end groups, resulting in a neutral or negative surface charge of the particle. Upon observation of cell growth for three days by live cell imaging using optical dark field microscopy, it was found that all particles supported cell adhesion while no directed cell migration and no significant particle internalization occurred. Concerning cell adhesion and spreading as compared to cell growth on bare substrates after 3 days of incubation, a reduction by 45% and 95%, respectively, for the surfactant particle coating was observed, whereas the amino-terminated polymer induced a reduction by 30% and 40%, respectively, which is absent for the carboxy-terminated polymer. Furthermore, interface-sensitive impedance spectroscopy (electric cell-substrate impedance sensing, ECIS) was employed in order to investigate the micromotility of cells added to substrates decorated with various amounts of surfactant-coated particles. A surface density of 65 particles/µm(2) (which corresponds to 0.5% of surface coverage with nanoparticles) diminishes micromotion by 25% as compared to bare substrates after 35 hours of incubation. We conclude that the surface coating of the gold nanorods, which were applied to the basolateral side of the cells, has a recognizable influence on the growth behavior and thus the coating should be carefully selected for biomedical applications of nanoparticles.

2.
J Am Chem Soc ; 135(15): 5620-30, 2013 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-23514537

RESUMO

The characterization of the lateral organization of components in biological membranes and the evolution of this arrangement in response to external triggers remain a major challenge. The concept of lipid rafts is widely invoked; however, direct evidence of the existence of these ephemeral entities remains elusive. We report here the use of secondary ion mass spectrometry (SIMS) to image the cholesterol-dependent cohesive phase separation of the ganglioside GM1 into nano- and microscale assemblies in a canonical lipid raft composition of lipids. This assembly of domains was interrogated in a model membrane system composed of palmitoyl sphingomyelin (PSM), cholesterol, and an unsaturated lipid (dioleoylphosphatidylcholine, DOPC). Orthogonal isotopic labeling of every lipid bilayer component and monofluorination of GM1 allowed generation of molecule specific images using a NanoSIMS. Simultaneous detection of six different ion species in SIMS, including secondary electrons, was used to generate ion ratio images whose signal intensity values could be correlated to composition through the use of calibration curves from standard samples. Images of this system provide the first direct, molecule specific, visual evidence for the colocalization of cholesterol and GM1 in supported lipid bilayers and further indicate the presence of three compositionally distinct phases: (1) the interdomain region; (2) micrometer-scale domains (d > 3 µm); (3) nanometer-scale domains (d = 100 nm to 1 µm) localized within the micrometer-scale domains and the interdomain region. PSM-rich, nanometer-scale domains prefer to partition within the more ordered, cholesterol-rich/DOPC-poor/GM1-rich micrometer-scale phase, while GM1-rich, nanometer-scale domains prefer to partition within the surrounding, disordered, cholesterol-poor/PSM-rich/DOPC-rich interdomain phase.


Assuntos
Colesterol/metabolismo , Gangliosídeo G(M1)/metabolismo , Espectrometria de Massa de Íon Secundário , Transporte Biológico , Colesterol/química , Gangliosídeo G(M1)/química , Bicamadas Lipídicas/química , Bicamadas Lipídicas/metabolismo , Microtecnologia , Modelos Moleculares , Conformação Molecular , Nanotecnologia
3.
Phys Rev E Stat Nonlin Soft Matter Phys ; 86(5 Pt 1): 051913, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23214820

RESUMO

The orchestration of cellular adhesion is an intricate process that involves a multitude of specialized proteins but at the same time follows a simple physical plan. It is still not totally known why cells create clusters of bonds to adhere on a substrate. Besides experimental methods to study cellular adhesion there exist several different theoretical models to describe the stability of focal adhesion clusters. The model proposed in this work makes it possible to understand the formation of adhesion bond clusters. It explains the emergence of a long-range bond-bond attraction that originates from the finite membrane elasticity as the driving force of cluster growth. In combination with a thermally driven bond disintegration such a model will eventually be able to explain the kinetic stability of finite size bond clusters. The model extends conventional two-state models where bonds are either open or closed by introducing a rich scenario of metastable states associated with hysteretic behavior in the bond closure and rupture dynamics.


Assuntos
Adesão Celular/fisiologia , Membrana Celular/química , Membrana Celular/fisiologia , Fluidez de Membrana/fisiologia , Modelos Biológicos , Modelos Químicos , Animais , Simulação por Computador , Humanos
4.
Small ; 8(23): 3683-90, 2012 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-22888068

RESUMO

Toxicological effects of nanoparticles are associated with their internalization into cells. Hence, there is a strong need for techniques revealing the interaction between particles and cells as well as quantifying the uptake at the same time. For that reason, herein optical dark-field microscopy is used in conjunction with transmission electron microscopy to investigate the uptake of gold nanoparticles into epithelial cells with respect to shape, stabilizing agent, and surface charge. The number of internalized particles is strongly dependent on the stabilizing agent, but not on the particle shape. A test of metabolic activity shows no direct correlation with the number of internalized particles. Therefore, particle properties besides coating and shape are suspected to contribute to the observed toxicity.


Assuntos
Ouro/química , Nanopartículas Metálicas/química , Animais , Cães , Células Epiteliais/metabolismo , Excipientes/química , Excipientes/metabolismo , Ouro/metabolismo , Células Madin Darby de Rim Canino , Microscopia Eletrônica de Transmissão , Tamanho da Partícula , Propriedades de Superfície
5.
J Am Chem Soc ; 134(7): 3326-9, 2012 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-22296574

RESUMO

The multivalent carbohydrate-carbohydrate interaction between membrane-anchored epitopes derived from the marine sponge Microciona prolifera has been explored by colloidal probe microscopy. An in situ coupling of sulfated and non-sulfated disaccharides to membrane-coated surfaces was employed to mimic native cell-cell contacts.The dynamic strength of the homomeric self-association was measured as a function of calcium ions and loading rate. A deterministic model was used to estimate the basic energy landscape and number of participating bonds in the contact zone.


Assuntos
Dissacarídeos/química , Bicamadas Lipídicas/química , Poríferos/química , Animais , Adesão Celular , Dissacarídeos/isolamento & purificação , Microscopia de Varredura por Sonda
6.
Chem Res Toxicol ; 24(9): 1494-506, 2011 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-21815656

RESUMO

Cellular motility is the major driving force of numerous biological phenomena including wound healing, immune response, embryogenesis, cancer formation, and metastasis. We studied the response of epithelial FaDu monolayers cultured on gold electrodes of an acoustic resonator (quartz crystal microbalance, QCM) and impedance sensor (electric cell-substrate impedance sensing, ECIS) to externally applied chemical stimuli interfering with cytoskeleton organization. Epithelial cell motility of confluent monolayers is characterized by subtle cell shape changes and variations in the cell-substrate as well as cell-cell distance without net directionality of individual cells. The impact of small molecules such as cytochalasin D, phalloidin, and blebbistatin as well as paclitaxel, nocodazol, and colchicin on actin and microtubules organization was quantified by conventional sensors' readouts and by comparing the noise pattern of the signals which is attributed to cellular dynamics. The responsiveness of noninvasive and label-free techniques relying on cellular dynamics is compared to classical viability assays and changes of the overall impedance of ultrasmall electrodes or acoustic loads of a thickness shear mode resonator. Depending on the agent used, a distinct sensor response was found, which can be used as a fingerprint of the cellular response. Cytoskeletal rearrangements and nuclear integrity were corroborated by fluorescence microscopy and correlated to the readouts of QCM and ECIS.


Assuntos
Técnicas Biossensoriais/métodos , Citoesqueleto/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Moduladores de Tubulina/toxicidade , Actinas/metabolismo , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Citocalasina D/toxicidade , Impedância Elétrica , Células Epiteliais/citologia , Compostos Heterocíclicos de 4 ou mais Anéis/toxicidade , Humanos , Paclitaxel/toxicidade , Faloidina/toxicidade , Técnicas de Microbalança de Cristal de Quartzo/métodos
7.
Nanotoxicology ; 5(2): 254-68, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21050076

RESUMO

Nanoparticle exposure is monitored by a combination of two label-free and non-invasive biosensor devices which detect cellular shape and viscoelasticity (quartz crystal microbalance), cell motility and the dynamics of epithelial cell-cell contacts (electric cell-substrate impedance sensing). With these tools we have studied the impact of nanoparticle shape on cellular physiology. Gold (Au) nanoparticles coated with CTAB were synthesized and studied in two distinct shapes: Spheres with a diameter of (43 ± 4) nm and rods with a size of (38 ± 7) nm × (17 ± 3) nm. Dose-response experiments were accompanied by conventional cytotoxicity tests as well as fluorescence and dark-field microscopy to visualize the intracellular particle distribution. We found that spherical gold nanoparticles with identical surface functionalization are generally more toxic and more efficiently ingested than rod-shaped particles. We largely attribute the higher toxicity of CTAB-coated spheres as compared to rod-shaped particles to a higher release of toxic CTAB upon intracellular aggregation.


Assuntos
Movimento Celular/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/fisiologia , Ouro/química , Nanopartículas Metálicas/toxicidade , Animais , Linhagem Celular , Cetrimônio , Compostos de Cetrimônio/química , Citoesqueleto/metabolismo , Relação Dose-Resposta a Droga , Células Epiteliais/citologia , Tamanho da Partícula , Espécies Reativas de Oxigênio/metabolismo , Propriedades de Superfície , Tensoativos/química
8.
Integr Biol (Camb) ; 2(2-3): 139-50, 2010 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-20473392

RESUMO

Early determination of the metastatic potential of cancer cells is a crucial step for successful oncological treatment. Besides the remarkable progress in molecular genomics- or proteomics-based diagnostics, there is a great demand for in vitro biosensor devices that allow rapid and selective detection of the invasive properties of tumor cells. Here, the classical cancer cell motility in vitro assays for migration and invasion relying on Boyden chambers are compared to a real-time biosensor that analyzes the dynamic properties of adherent cells electro-acoustically with a time resolution on the order of seconds. The sensor relies on the well-established quartz crystal microbalance technique (QCM) that measures the shift in resonance frequency and damping of an oscillating quartz crystal when adsorption, desorption or changes in material properties close to the quartz surface occur. In addition, the QCM is capable of detecting the rather subtle fluctuations of the cell bodies as an indicator for their micromotility. QCM-based micromotility readings of three different cancer cell lines (HT-29, HSC-4, FaDu) are compared with the well-known electrical cell-substrate impedance sensing (ECIS) revealing collective stochastic motion that corresponds to the malignancy of the cells.


Assuntos
Acústica/instrumentação , Técnicas Biossensoriais/instrumentação , Sistemas Microeletromecânicos/instrumentação , Neoplasias/patologia , Neoplasias/fisiopatologia , Linhagem Celular Tumoral , Humanos
9.
Biophys Chem ; 150(1-3): 54-63, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20219280

RESUMO

A versatile model system to study membrane-membrane interactions in great detail is introduced. Based on colloidal probe microscopy with membrane covered spherical probes attached to tip-less cantilevers the interaction forces and adhesion energies are quantified down to single molecule resolution. Two opposing membranes equipped with ligands on one side and receptors on the other side were brought in contact at a defined load and pulled apart at constant velocity. Ni-NTA functionalized lipids served as receptors on the probe, while lipopeptides displaying short His-tags (CGGH(6) or CGWH(6)) were incorporated in the planar supporting membrane on a silicon substrate. The rather intricate force distance curves were scrutinized in terms of breakthrough events upon contact of the probe with the surface, the overall work of adhesion, maximum adhesion force, as well as formation frequency, lifetime, and force of membrane tethers suggesting that hemifusion of the two opposing bilayers takes place.


Assuntos
Membrana Celular/metabolismo , Lipídeos de Membrana/metabolismo , Microscopia de Varredura por Sonda/métodos , Ligantes , Bicamadas Lipídicas/química , Bicamadas Lipídicas/metabolismo , Lipopeptídeos/química , Lipopeptídeos/metabolismo , Lipossomos/química , Lipossomos/metabolismo , Fusão de Membrana , Lipídeos de Membrana/química , Microscopia de Varredura por Sonda/instrumentação , Níquel/química , Níquel/metabolismo , Ácido Nitrilotriacético/química , Ácido Nitrilotriacético/metabolismo , Fosfolipídeos/química , Fosfolipídeos/metabolismo , Fosforilcolina/química , Fosforilcolina/metabolismo , Silício/química
10.
ACS Nano ; 3(1): 213-22, 2009 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-19206269

RESUMO

In the growing field of nanotechnology, there is an urgent need to sensitively determine the toxicity of nanoparticles since many technical and medical applications are based on controlled exposure to particles, that is, as contrast agents or for drug delivery. Before the in vivo implementation, in vitro cell experiments are required to achieve a detailed knowledge of toxicity and biodegradation as a function of the nanoparticles' physical and chemical properties. In this study, we show that the micromotility of animal cells as monitored by electrical cell-substrate impedance analysis (ECIS) is highly suitable to quantify in vitro cytotoxicity of semiconductor quantum dots and gold nanorods. The method is validated by conventional cytotoxicity testing and accompanied by fluorescence and dark-field microscopy to visualize changes in the cytoskeleton integrity and to determine the location of the particles within the cell.


Assuntos
Metais/química , Nanopartículas/química , Nanotecnologia/métodos , Pontos Quânticos , Animais , Biodegradação Ambiental , Meios de Contraste/farmacologia , Sistemas de Liberação de Medicamentos , Impedância Elétrica , Corantes Fluorescentes/química , Ouro/química , Humanos , Nanopartículas Metálicas/química , Semicondutores
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...