Nanobiotechnology and Cell Biology: Micro- and Nanofabricated Surfaces to Investigate Receptor-Mediated Signaling

Torres, Alexis J.; Wu, Min; Holowka, David; Baird, Barbara

doi:10.1146/annurev.biophys.36.040306.132651

Cited by 91 publications

(108 citation statements)

References 90 publications

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“…In spite of the porosity of the electrospun mats, they must be considered as a two dimensional substrates with respect to cell culture since inter-fiber spaces do not allow cell invasion of the three dimensional fiber structure. Cell-substrate interaction mediated by proteins adsorbed from the culture medium is crucial to control chondrocyte adhesion, proliferation and consequently gene expression in monolayer culture [5,[47][48]. It has been shown that nano-fibrilar substrates allow viable and proliferative chondrocyte culture [32,49].…”

Section: Discussionmentioning

confidence: 99%

Influence of crystallinity and fiber orientation on hydrophobicity and biological response of poly(l-lactide) electrospun mats

et al. 2012

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Royal Society of ChemistryAreias, A.; Ribeiro, C.; Sencadas, V.; Garcia Giralt, N.; Diez Perez, A.; Gómez Ribelles, JL.; Lanceros Mendez, S. (2012). Influence of crystallinity and fiber orientation on hydrophobicity and biological response of poly(L-lactide) electrospun mats. Soft Matter. 8 (21) AbstractPoly(L-lactide) electrospun mates have been produced with random and aligned fiber orientation and degrees of crystallinity from 0 up to nearly 50%. These two factors, fiber alignment and degree of crystallinity strongly affect the hydrophobicity of the samples, being this larger for the aligned fiber mats and for the fibers with higher degree of crystallinity. Whereas the first effect can be associated to a decrease in the degree of porosity the second should be related to variations in the fiber roughness at nanometric scale and an increase in fiber stiffness. Proliferation of human chondrocytes cultured in monolayer on these substrates is similar in both aligned and non-aligned amorphous mats. Crystallization of the aligned mats, on the other hand, nearly suppresses proliferation and the cells produce higher amounts of aggrecan, characteristic of the extracellular matrix of hyaline cartilage.

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Section: Discussionmentioning

confidence: 99%

Influence of crystallinity and fiber orientation on hydrophobicity and biological response of poly(l-lactide) electrospun mats

et al. 2012

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“…We and others demonstrated the versatility of patterned ligands that confine and activate receptors in micrometer-size regions so that the spatially regulated cellular responses can be monitored (for review, see ref. 16). An important feature of patterned lipid bilayers is that proteins do not adhere nonspecifically to the lipid regions: anti-DNP IgE-Fc RI binds selectively to these regions only because DNP-cap-PE is incorporated in the bilayers (17).…”

Section: Discussionmentioning

confidence: 99%

Focal adhesion proteins connect IgE receptors to the cytoskeleton as revealed by micropatterned ligand arrays

Torres

Vasudevan

Holowka

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

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Patterned surfaces that present specific ligands in spatially defined arrays are used to examine structural linkages between clustered IgE receptors (IgE-Fc RI) and the cytoskeleton in rat basophilic leukemia (RBL) mast cells. We showed with fluorescence microscopy that cytoskeletal F-actin concentrates in the same regions as cell surface IgE-Fc RI that bind to the micrometer-size patterned ligands. However, the proteins mediating these cytoskeletal connections and their functional relevance were not known. We now show that whereas the adaptor proteins ezrin and moesin do not detectably concentrate with the array of clustered IgE-Fc RI, focal adhesion proteins vinculin, paxillin, and talin, which are known to link F-actin with integrins, accumulate in these regions on the same time scale as F-actin. Moreover, colocalization of these focal adhesion proteins with clustered IgE-Fc RI is enhanced after addition of fibronectin-RGD peptides. Significantly, the most prominent rat basophilic leukemia cell integrin (␣5) avoids the patterned regions occupied by the ligands and associates preferentially with exposed regions of the silicon substrate. Thus, spatial separation provided by the patterned surface reveals that particular focal adhesion proteins, which connect to the actin cytoskeleton, associate with ligand-cross-linked IgE-Fc RI, independently of integrins. We investigated the functional role of one of these proteins, paxillin, in IgE-Fc RI-mediated signaling by using small interfering RNA. From these results, we determine that paxillin reduces stimulated phosphorylation of the Fc RI ␤ subunit but enhances stimulated Ca 2؉ release from intracellular stores. The results suggest that paxillin associated with clustered IgE-Fc RI has a net positive effect on Fc RI signaling.FceRI ͉ Lyn ͉ nanobiotechnology ͉ paxillin ͉ supported lipid bilayers C ells have evolved to respond to their chemical and physical environment. Chemical stimulants in the form of soluble or surface-bound ligands are recognized by specific cell surface receptors, and physical cues are sensed by integrins that bind to extracellular matrix proteins on surrounding substrates (1). Cross-talk between intracellular signaling pathways that are initiated by integrins and by ligand receptors has been clearly demonstrated, although spatial aspects of these processes have not been defined. Surfaces patterned on the micrometer scale offer the opportunity to separate regions that bind integrins from those that present ligands to cell surface receptors and thereby delineate respective cytoskeletal connections.The subject of our study is the IgE receptor (Fc RI), which is a member of the family of multisubunit immune recognition receptors that includes antigen receptors on B cells and T cells. This family of receptors has conserved structural features and similarly initiates intracellular signaling in response to multivalent ligands (antigens) that activate cells by clustering cell surface receptors. Fc RI receptors are found primarily in mast cells and basophi...

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“…Another great advantage of the FCS is the possibility to accurately quantify local measurements of molecular mobility on diVerent temporal scales. Among the diVerent nanotechnologies applied on biomembranes (Torres et al 2008), the ones combining single nanometric aperture technology with FCS have great potential as recently demonstrated (Edel et al 2005;Moran-Mirabal et al 2007;Samiee et al 2006;Wenger et al 2006b). Indeed, these approaches preserve the FCS temporal resolution while providing a subwavelength spatial resolution.…”

Section: Biological Applicationsmentioning

confidence: 99%

Fluorescence fluctuations analysis in nanoapertures: physical concepts and biological applications

Lenne¹,

Rigneault²,

Marguet³

et al. 2008

Histochem Cell Biol

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During the past years, nanophotonics has provided new approaches to study the biological processes below the optical diVraction limit. How single molecules diVuse, bind and assemble can be studied now at the nanometric level, not only in solutions but also in complex and crowded environments such as in live cells. In this context Xuorescence Xuctuations spectroscopy is a unique tool since it has proven to be easy to use in combination with nanostructures, which are able to conWne light in nanometric volumes. We review here recent advances in Xuorescence Xuctuations' analysis below the optical diVraction limit with a special focus on nanoapertures milled in metallic Wlms. We discuss applications in the Weld of single-molecule detection, DNA sequencing and membrane organization, and underscore some potential perspectives of this new emerging technology.

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Nanobiotechnology and Cell Biology: Micro- and Nanofabricated Surfaces to Investigate Receptor-Mediated Signaling

Cited by 91 publications

References 90 publications

Influence of crystallinity and fiber orientation on hydrophobicity and biological response of poly(l-lactide) electrospun mats

Influence of crystallinity and fiber orientation on hydrophobicity and biological response of poly(l-lactide) electrospun mats

Focal adhesion proteins connect IgE receptors to the cytoskeleton as revealed by micropatterned ligand arrays

Fluorescence fluctuations analysis in nanoapertures: physical concepts and biological applications

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