Label-free optical monitoring of surface adhesion of extracellular vesicles by grating coupled interferometry

Patko, Daniel; György, Bence; Németh, Andrea H.; Szabó-Taylor, Katalin; Kittel, Ágnes; Buzás, Edit I.; Horváth, Róbert

doi:10.1016/j.snb.2013.07.035

Cited by 33 publications

(24 citation statements)

References 22 publications

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“…Conditioned tissue culture supernatant of CCRF cells was centrifuged 300g 10 minutes, the pellet was discarded, and the supernatant was submitted to resistive pulse sensing analysis using a qNano instrument (IZON Science Ltd, New Zeland) as described before [13].…”

Section: Determination Of Ev Concentration and Size Distribution Usinmentioning

confidence: 99%

Critical role of extracellular vesicles in modulating the cellular effects of cytokines

Szabó

Tarr

Pálóczi

et al. 2014

Cell. Mol. Life Sci.

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Under physiological and pathological conditions, extracellular vesicles (EVs) are present in the extracellular compartment simultaneously with soluble mediators. We hypothesized that cytokine effects may be modulated by EVs, the recently recognized conveyors of intercellular messages. In order to test this hypothesis, human monocyte cells were incubated with CCRF acute lymphoblastic leukemia cell line-derived EVs with or without the addition of recombinant human TNF, and global gene expression changes were analysed. EVs alone regulated the expression of numerous genes related to inflammation and signaling. In combination, the effects of EVs and TNF were additive, antagonistic or independent. The differential effects of EVs and TNF or their simultaneous presence were also validated by Taqman assays, ELISA and testing different populations of purified EVs. In the case of the paramount chemokine IL-8, we were able to demonstrate a synergistic up-regulation by purified EVs and TNF. Our data suggest that neglecting the modulating role of EVs on the effects of soluble mediators may skew experimental results. On the other hand, considering the combined effects of cytokines and EVs may prove therapeutically useful by targeting both compartments at the same time.

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Section: Determination Of Ev Concentration and Size Distribution Usinmentioning

confidence: 99%

Critical role of extracellular vesicles in modulating the cellular effects of cytokines

Szabó

Tarr

Pálóczi

et al. 2014

Cell. Mol. Life Sci.

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“…Evanescent field-based optical biosensors, including surface plasmon resonance (SPR), 36,37 optical waveguide lightmode spectroscopy (OWLS), 28,[38][39][40] photonic crystal biosensors, 41 grating coupling interferometry, 42,43 and resonant waveguide grating (commonly recognized as Epic) 44,45 biosensors, are considered to be especially straightforward means to monitor surface adhesion. First, they are sensitive only in a $150 nm thick layer closest to the substratum, exactly where establishment and maturation of anchorage and spreading occurs.…”

Section: Introductionmentioning

confidence: 99%

Adhesion kinetics of human primary monocytes, dendritic cells, and macrophages: Dynamic cell adhesion measurements with a label-free optical biosensor and their comparison with end-point assays

et al. 2016

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Monocytes, dendritic cells (DCs), and macrophages (MFs) are closely related immune cells that differ in their main functions. These specific functions are, to a considerable degree, determined by the differences in the adhesion behavior of the cells. To study the inherently and essentially dynamic aspects of the adhesion of monocytes, DCs, and MFs, dynamic cell adhesion assays were performed with a high-throughput label-free optical biosensor [Epic BenchTop (BT)] on surfaces coated with either fibrinogen (Fgn) or the biomimetic copolymer PLL-g-PEG-RGD. Cell adhesion profiles typically reached their maximum at ∼60 min after cell seeding, which was followed by a monotonic signal decrease, indicating gradually weakening cell adhesion. According to the biosensor response, cell types could be ordered by increasing adherence as monocytes, MFs, and DCs. Notably, all three cell types induced a larger biosensor signal on Fgn than on PLL-g-PEG-RGD. To interpret this result, the molecular layers were characterized by further exploiting the potentials of the biosensor: by measuring the adsorption signal induced during the surface coating procedure, the authors could estimate the surface density of adsorbed molecules and, thus, the number of binding sites potentially presented for the adhesion receptors. Surfaces coated with PLL-g-PEG-RGD presented less RGD sites, but was less efficient in promoting cell spreading than those coated with Fgn; hence, other binding sites in Fgn played a more decisive role in determining cell adherence. To support the cell adhesion data obtained with the biosensor, cell adherence on Fgn-coated surfaces 30–60 min after cell seeding was measured with three complementary techniques, i.e., with (1) a fluorescence-based classical adherence assay, (2) a shear flow chamber applying hydrodynamic shear stress to wash cells away, and (3) an automated micropipette using vacuum-generated fluid flow to lift cells up. These techniques confirmed the results obtained with the high-temporal-resolution Epic BT, but could only provide end-point data. In contrast, complex, nonmonotonic cell adhesion kinetics measured by the high-throughput optical biosensor is expected to open a window on the hidden background of the immune cell–extracellular matrix interactions.

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“…Waveguide sensors were recently exploited in several novel directions; such as the on-line monitoring of oriented protein adsorption [41], extracellular vesicles [42] and human primary immune [16] and cancer [17] cell adhesion in a completely labelfree manner. Here we report on the preparation of TNT coatings on surface-sensitive optical waveguide-based biosensors for quantitative adsorption and adhesion studies.…”

Section: Introductionmentioning

confidence: 99%

Enhanced protein adsorption and cellular adhesion using transparent titanate nanotube thin films made by a simple and inexpensive room temperature process: Application to optical biochips

Nádor

Orgován

Fried

et al. 2014

Colloids and Surfaces B: Biointerfaces

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Please cite this article in press as: J. Nador, et al., Enhanced protein adsorption and cellular adhesion using transparent titanate nanotube thin films made by a simple and inexpensive room temperature process: Application to optical biochips, Colloids Surf. a b s t r a c tA new type of titanate nanotube (TNT) coating is investigated for exploitation in biosensor applications. The TNT layers were prepared from stable but additive-free sols without applying any binding compounds. The simple, fast spin-coating process was carried out at room temperature, and resulted in well-formed films around 10 nm thick. The films are highly transparent as expected from their nanostructure and may, therefore, be useful as coatings for surface-sensitive optical biosensors to enhance the specific surface area. In addition, these novel coatings could be applied to medical implant surfaces to control cellular adhesion. Their morphology and structure was characterized by spectroscopic ellipsometry (SE) and atomic force microscopy (AFM), and their chemical state by X-ray photoelectron spectroscopy (XPS). For quantitative surface adhesion studies, the films were prepared on optical waveguides. The coated waveguides were shown to still guide light; thus, their sensing capability remains. Protein adsorption and cell adhesion studies on the titanate nanotube films and on smooth control surfaces revealed that the nanostructured titanate enhanced the adsorption of albumin; furthermore, the coatings considerably enhanced the adhesion of living mammalian cells (human embryonic kidney and preosteoblast).

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Label-free optical monitoring of surface adhesion of extracellular vesicles by grating coupled interferometry

Abstract: In this proof-of-principle study a label-free optical sensor is demonstrated to monitor the surface adhesion of extracellular vesicles secreted by live cells on to various extracellular matrix proteins.

Cited by 33 publications

References 22 publications

Critical role of extracellular vesicles in modulating the cellular effects of cytokines

Critical role of extracellular vesicles in modulating the cellular effects of cytokines

Adhesion kinetics of human primary monocytes, dendritic cells, and macrophages: Dynamic cell adhesion measurements with a label-free optical biosensor and their comparison with end-point assays

Enhanced protein adsorption and cellular adhesion using transparent titanate nanotube thin films made by a simple and inexpensive room temperature process: Application to optical biochips

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