RESUMO
We investigate the vibrational modes of gold nanorings on a silica substrate with an ultrafast optical technique. By comparison with numerical simulations, we identify several resonances in the gigahertz range associated with axially symmetric deformations of the nanoring and substrate. We elucidate the corresponding mode shapes and find that the substrate plays an important role in determining the mode damping. This study demonstrates the need for a plasmonic nano-optics approach to understand the optical excitation and detection mechanisms for the vibrations of plasmonic nanostructures.
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A simple approach to enhance the refractive index sensitivity of gold nanodisks immobilized on electrically conducting indium tin oxide (ITO) substrates has been demonstrated. A two-fold increase in sensitivity to bulk refractive index change was achieved by substrate under-etching of gold nanodisks on ITO in 50 mM sulfuric acid. The influence of an intermediate titanium adhesion layer was investigated and was found to markedly influence the etching pattern and time. Etching with an adhesion layer resulted in enhanced refractive index sensitivity on disk-on-pin like structures after long etching times, whereas etching of disks deposited directly on ITO resulted in a disk-on-pincushion like configuration and similarly enhanced sensitivity already at shorter times. The gold disks remained electrically connected to the ITO substrate throughout etching and allowed site-specific electrodeposition of poly(3-aminophenol) at the nanodisks, showing enhanced thin-film refractive index sensitivity. This work demonstrates a simple method for enhancing refractive index sensitivity of nanostructures on ITO substrates for combined electrochemical and optical platforms, and subsequently a method to modify the surface of the electrically connected nanostructures, which has potential application in biosensing.
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The separate influence of topographical and chemical cues on cell attachment and spreading are well documented; however, that of duel-cue substrates is less so. In this study graft copolymers that sterically stabilize biological surfaces were employed alongside nanotopographical features fabricated by colloidal lithography. This resulted in the production of a range of substrates whereby the effect of chemistry and or topography on both on human fibroblast and bone marrow cell adhesion and spreading could be observed. The current studies indicate an enhancement of cell response as a consequence of modifications in material topography, whereas the current selected chemical cues inhibited cell function. Critically, in combination, topography modulated the effects of chemical environment.
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Células da Medula Óssea/metabolismo , Fibroblastos/metabolismo , Nanoestruturas/química , Nanoestruturas/ultraestrutura , Proteínas/química , Proteínas/metabolismo , Engenharia Tecidual/métodos , Materiais Biocompatíveis/química , Células da Medula Óssea/citologia , Adesão Celular/fisiologia , Técnicas de Cultura de Células/métodos , Proliferação de Células , Sobrevivência Celular , Células Cultivadas , Fibroblastos/citologia , Humanos , Teste de Materiais , Especificidade por Substrato , Propriedades de SuperfícieRESUMO
In this paper, we utilize colloidal lithography based on electrostatic self-assembly of polystyrene colloidal particles onto a polymer surface as a nanoscale mask. The pattern is then transferred to the surface by ion beam etching. Each particle acts as an individual mask, resulting in an array of identical structure. Ion beam exposure etches away the unmasked surface between the particles, so the particle mask pattern can be transferred into the polymer surface. This method allows to nanofabricate bulk polymeric surfaces with systematic variation in relief, structure sizes, and aspect ratios. It is a fast, simple, and reliable method to fabricated different polymeric surfaces even on large area samples (> 1 cm2). The structural variation is achieved by use of different conditions during the self-assembly of the mask (e.g., different particles sizes) or different ion etching conditions during the pattern transfer (e.g., ion energy, ion flux, ion incident angle, etching time, gas environment).
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Materiais Biocompatíveis/química , Coloides/química , Nanoestruturas/química , Nanoestruturas/ultraestrutura , Nanotecnologia/métodos , Fotografação/métodos , Polimetil Metacrilato/química , Íons , Teste de Materiais , Tamanho da Partícula , Polímeros/química , Propriedades de SuperfícieRESUMO
In recent years the progress in the field of nanotechnologies has offered new possibilities to control the superficial features of implant materials down to a nanoscale level. Several studies have therefore tried to explore the effects of nanostructured biomaterial surfaces on the behavior of eukaryotic cells. However, nanotopography could exert an influence also on the behavior of prokaryotic cells, with relevant implications concerning the susceptibility of implant surfaces to infection. Aim of this study was to examine the behavior of Staphylococcus aureus on polyethylene terephthalate (PET) surfaces either cylindrically nanostructured (PET-N) or flat ion-etched (PET-F), and on tissue culture-grade polystyrene (PS). Microbial adherence was assessed by chemiluminometry under 4 different conditions: (a) bacteria suspended in MEM medium, (b) bacteria in MEM supplemented with 10% fetal bovine serum (FBS), (c) test surfaces preconditioned in FBS, and (d) post-exposure of colonised surfaces to serum-supplemented MEM. Under all circumstances, PET-F and PET-N specimens showed identical bacterial adhesion properties. In the absence of serum, all 3 test materials showed a very high adhesivity to microbial cells and both PET surfaces exhibited greater adhesion than PS. On the contrary, the presence of 10% serum in solution significantly affected cell behavior: the number of microbial cells on all surfaces was drastically reduced, and the adhesion properties of PET surfaces with respect to PS were reversed, with PET being less adhesive. Overall, the specific cylindrical nanostructures created on PET did not significantly influence microbial behavior. Ongoing studies are verifying whether other nanotopographies with different geometry could have more substantial effects.
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Aderência Bacteriana/fisiologia , Staphylococcus aureus/fisiologia , Materiais Biocompatíveis , Contagem de Colônia Microbiana , Meios de Cultura , Meios de Cultura Livres de Soro , Medições Luminescentes , Teste de Materiais , Nanopartículas , Nanotecnologia , Polietilenotereftalatos , Poliestirenos , Propriedades de SuperfícieRESUMO
The environment around a cell during in vitro culture is unlikely to mimic those in vivo. Preliminary experiments with nanotopography have shown that nanoscale features can strongly influence cell morphology, adhesion, proliferation and gene regulation, but the mechanisms mediating this cell response remain unclear. In this study a well defined nanotopography, consisting of 100 nm wide and 160 nm high cylindrical columns, was used in fibroblast culture. In order to build on previously published morphological data that showed changes in cell spreading on the nanocolumns, in this study gene regulation was monitored using a 1718 gene microarray. Transmission electron microscopy, fluorescent observation of actin and Rac and area quantification have been used to re-affirm the microarray observations. The results indicate that changes in cell spreading correlate with a number of gene up- and down-regulations as will be described within the manuscript.
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Coloides , Fibroblastos/citologia , Análise em Microsséries/métodos , Nanotecnologia/instrumentação , Animais , Células Cultivadas , Regulação para Baixo/genética , Fibroblastos/ultraestrutura , Humanos , Microscopia de Força Atômica , Regulação para Cima/genéticaRESUMO
A novel approach combining self-assembly-based colloidal lithography and polydimethylsiloxane (PDMS) micromolding to generate complex protein nanopatterns for studying the mechanisms of leukocyte extravasation within microchannels is presented. Nanostructured surfaces sealed onto PDMS-molded microchannels are chemically functionalized in situ in an all-aqueous process to generate bi-functional chemical nanopatterns. Subsequent co-immobilization with proteins makes use of common non-covalent coupling (e.g. HIS-tags, FC-tags and biotin-tags), giving nanopatterns of arbitrary combinations of oriented, functional proteins. Up to three different proteins were simultaneously co-immobilized into the microchannel with nanoscale precision, demonstrating the complex patterns. As a proof-of-principle, a mimic of an inflamed endothelium was constructed using a macro- and nanoscale pattern of intercellular adhesion molecule 1 (ICAM1) and P-selectin, and the response of leukocytes through live cell imaging was measured. A clear result on the rolling behavior of the cells was observed with rolling limited to areas where ICAM1 and P-selectin are present. This micro/nano-interface will open new doors to investigations of how spatial distributions of proteins control cellular activity.
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Bioimpressão/instrumentação , Proteínas Imobilizadas/química , Molécula 1 de Adesão Intercelular/química , Leucócitos/citologia , Técnicas Analíticas Microfluídicas/instrumentação , Nanoestruturas/química , Selectina-P/química , Linhagem Celular , Dimetilpolisiloxanos/química , Desenho de Equipamento , Humanos , Migração e Rolagem de LeucócitosRESUMO
Medical grade titanium samples were examined using X-ray photoelectron spectroscopy before and after immersion in various proteins. Additionally, an implant removed from a patient following clinical failure was examined using scanning ion and electron microscopy. The surface of the as-received samples was found to be mainly TiO2, with contaminants of H2O/OH-, calcium and nitrogen which remained after autoclaving. The immersed proteins adhered to the titanium surface, possibly via a Ca-O link. The failed clinical sample was found to be partially fibrously encapsulated with evidence of calcification. Small amounts of TiOOH were detected at the fibrous periphery, supporting the theory of Tengvall and co-workers that in vivo titanium implants are covered in a gel of this material.
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Próteses e Implantes , Titânio/química , Osso e Ossos/fisiologia , Microanálise por Sonda Eletrônica , Glicosaminoglicanos/química , Humanos , Mandíbula , Microscopia Eletrônica de Varredura , Falha de Prótese , Propriedades de SuperfícieRESUMO
The effect of nanotopography on a range of Ti oxide surfaces was determined. Flat Ti, 3%, 19%, 30% and 43% topography densities of 110 nm high hemispherical protrusions were cultured in contact with primary derived human macrophages and osteoblasts in single culture models. Prior to introduction of the test substrate the phenotype and optimum conditions for in vitro cell culture were established. The cellular response was investigated and quantified by assessments of cytoskeletal development and orientation, viable cell adhesion, cytokine production and release and RT-PCR analysis of osteogenic markers. The tested nanotopographies did not have a statistically significant effect on viable cell adhesion and subsequent cytoskeletal formation. Surface chemistry was the dominant factor as established via incorporation of a tissue culture polystyrene, TCPS, control. The topography surfaces induced a release of chemotactic macrophage activation agents at 1 day in conjunction with stress fibre formation and a subsequent fibronectin network formation. Osteoblasts migrated away from the topography surfaces to the exposed TCPS within the wells during the 7-day period.
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Técnicas de Cultura de Células/métodos , Materiais Revestidos Biocompatíveis , Citocinas/metabolismo , Macrófagos/citologia , Macrófagos/fisiologia , Teste de Materiais , Osteoblastos/citologia , Osteoblastos/fisiologia , Adesão Celular/fisiologia , Divisão Celular/fisiologia , Movimento Celular/fisiologia , Polaridade Celular/fisiologia , Sobrevivência Celular/fisiologia , Células Cultivadas , Citoesqueleto/fisiologia , Citoesqueleto/ultraestrutura , Humanos , Nanotubos , Propriedades de Superfície , TitânioRESUMO
Large area nanopatterns of functional proteins are demonstrated. A new approach to analyze atomic force microscopy height histograms is used to quantify protein and antibody binding to nanoscale patches. Arrays of nanopatches, each containing less than 40 laminin molecules, are shown to be highly functional binding close to 1 monoclonal anti-laminin IgG (site by IKVAV sequence) or 3-4 polyclonal anti-laminin IgG's per surface bound laminin. Complementary quartz crystal microbalance measurements indicate higher functionality at nanopatches than on homogeneous surfaces.
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Cristalização/métodos , Laminina/química , Laminina/ultraestrutura , Nanoestruturas/química , Nanoestruturas/ultraestrutura , Adsorção , Sítios de Ligação , Biologia/métodos , Materiais Revestidos Biocompatíveis/química , Teste de Materiais , Microscopia de Força Atômica , Tamanho da Partícula , Ligação Proteica , Propriedades de SuperfícieRESUMO
The optical response of ring-shaped gold nanoparticles prepared by colloidal lithography is investigated. Compared to solid gold particles of similar size, nanorings exhibit a redshifted localized surface plasmon that can be tuned over an extended wavelength range by varying the ratio of the ring thickness to its radius. The measured wavelength variation is well reproduced by numerical calculations and interpreted as originating from coupling of dipolar modes at the inner and outer surfaces of the nanorings. The electric field associated with these plasmons exhibits uniform enhancement and polarization in the ring cavity, suggesting applications in near-infrared surface-enhanced spectroscopy and sensing.