Communication between hydrogel beads<i>via</i>chemical signalling

Jaggers, Ross W.; Bon, Stefan Antonius Franciscus

doi:10.1039/c7tb02278f

Cited by 7 publications

(8 citation statements)

References 36 publications

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“…This delivery system is based on the use of hydrogel agarose microscale beads which we utilize to encapsulate the cell – free lysate and couple with transcription and translation (TXTL) processes (SI Figure 1). The size of the final microscale beads can be adjusted from diameters of 10 μm – 100 μm with minor adjustments to flow and pressure to the microfluidic processing and the porosity is an innate property of the cross-linking density of the agarose which can be controlled by agarose concentration [32] [33]. The resulting cell – free hydrogel device is a robust biosensor that can be delivered to both biologically and chemically hostile environments for controlled sensing and signaling of Pseudomonas infection.…”

Section: Resultsmentioning

confidence: 99%

On a Robust, Sensitive Cell – Free Method for Pseudomonas Sensing and Quantification

Seto

Buske

Laurie

2018

Preprint

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Cell -free lysates have found a utility in performing cellular functions and providing biologically-relevant metabolic products without the optimal biological conditions for growth and proliferation. By teasing out the biological components and constructing artificial conditions that enable for biological transcription and translation processes to occur, specific cell -like functions can be reconstituted in vitro without requiring the entire cell and milieu of cellular organelles. This enables for optimization of biological circuits, either by concentration or on/off switches, simply through the addition or removal of genetic components (plasmids, inducers, or repressors) of regulatory elements. Here, we demonstrate an application of cell -free process that is robust and portable, independent of a substrate, to apply for sensing and reporting functions of a quorum sensing molecule N-3-oxododecanoyl homoserine lactone (3OC12HSL) found crucial for pathological Pseudomonas infections. Using droplet microfluidics to integrate cell -free related functions into hydrogel scaffolds, we show that these cell -free circuits can be encapsulated, delivered, and activated in most environments-even in conditions with very little hydration. IntroductionBiological engineering of organisms and natural systems have ushered novel methods to examine disease and dysfunction in vivo [1][2][3][4][5]. Recent innovations using cell -free lysates for various functions have led to a variety of applications from biocomputation [6, 7] molecular design and engineering [8, 9], metabolic pathway engineering [10][11][12] to novel methods of protein synthesis [13][14][15]. In all these examples of cell -free lysate applications, the underlying premise of transcription and translation is still relied upon [16, 17]; however, with modifications and the addition of synthetic energy pathways, these cellular processes can be engineered to operate ex vivo.

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Section: Resultsmentioning

confidence: 99%

On a Robust, Sensitive Cell – Free Method for Pseudomonas Sensing and Quantification

Seto

Buske

Laurie

2018

Preprint

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“…This delivery system is based on the use of hydrogel agarose microscale beads which we utilize to encapsulate the cell-free lysate and couple with transcription and translation (TXTL) processes (Figure 2). The size of the final microscale beads can be adjusted from diameters of 10–100 µm with minor adjustments to flow and pressure to the microfluidic processing and the porosity is an innate property of the cross-linking density of the agarose which can be controlled by agarose concentration [38,39]. Unlike other findings, the use of agarose is not toxic for the LasR quorum-sensing mechanism [28,40].…”

Section: Discussionmentioning

confidence: 99%

On a Robust, Sensitive Cell-Free Method for Pseudomonas Sensing and Quantification in Microfluidic Templated Hydrogels

Seto

2019

Micromachines

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Through the use of droplet microfluidics to integrate cell-free activity into inert hydrogel beads, we have developed a platform that can perform biologically relevant functions without the need for cells. Specifically, cell-free lysates serve a utility in performing cellular functions and providing biologically relevant metabolic products without requiring the optimal biological conditions for cell growth and proliferation. By teasing out specific biological components that enable transcription and translation to occur, these cell-like functions can be reconstituted in vitro without requiring the entire cell and milieu of cellular organelles. This enables the optimization of synthetic biological circuits, either by concentration or logic switches, simply through the addition or removal of genetic components (plasmids, inducers, or repressors) of regulatory elements. Here, we demonstrate an application of cell-free processes that is robust and portable, independent of a substrate, to apply for sensing and reporting functions of a quorum-sensing molecule N-3-oxododecanoyl homoserine lactone (3OC12HSL) found crucial for pathological Pseudomonas aeruginosa infection. We develop an agarose bead platform that is easily adaptable and simply programmable to fit a variety of biological and chemical sensing applications for the utility of ease of delivery and activation in remote environments—even in conditions with very little hydration.

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“…There has also been interest in using pH changes in enzyme-loaded polymer gels to achieve spatial and temporal control. In 2017, Jaggers and Bon used pH changes induced by urease-loaded calcium alginate gels to change the color, or induce the disassembly of gel structures . Taylor, Pojman and co-workers used this urease-driven base-releasing hydrogel to initiate a polymerization process in close proximity to the hydrogel .…”

Section: Introductionmentioning

confidence: 99%

Fabricating Shaped and Patterned Supramolecular Multigelator Objects via Diffusion-Adhesion Gel Assembly

Tangsombun,

Smith

2023

J. Am. Chem. Soc.

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We report the use of acid-diffusion to assemble core–shell supramolecular gel beads with different low-molecular-weight gelators (LMWGs) in the core and shell. These gel beads grow a shell of dibenzylidenesorbitol-based DBS-COOH onto a core comprising DBS-CONHNH2 and agarose that has been loaded with acetic acid. Diffusion of the acid from the core triggers shell assembly. The presence of DBS-CONHNH2 enables the gel core to be loaded with metal nanoparticles (NPs) as acyl hydrazide reduces metal salts in situ. The pH-responsiveness of DBS-COOH allows responsive assembly of the shell with both temporal and spatial control. By fixing multiple gel beads in a Petri dish, the cores become linked to one another by the assembled DBS-COOH gel shella process we describe as diffusion-adhesion assembly. By controlling the geometry of the beads with respect to one another, it is possible to pattern the structures, and using a layer-by-layer approach, 3D objects can be fabricated. If some of the beads are loaded with basic DBS-carboxylate instead of CH3COOH, they act as a “sink” for diffusing protons, preventing DBS-COOH shell assembly in the close proximity. Those beads do not adhere to the remainder of the growing gel object and can be simply removed once diffusion-assembly is complete, acting as templates, and enabling the fabrication of 3D “imprinted” multigel architectures. Preloading the gel beads with AuNPs or AgNPs suspends these functional units within the cores at precisely defined locations within a wider gel object. In summary, this approach enables the dynamic fabrication of shaped and patterned gels with embedded metal NPssuch objects have potential next-generation applications in areas including soft nanoelectronics and regenerative medicine.

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Communication between hydrogel beadsviachemical signalling

Abstract: In this work, we demonstrate chemical communication between millimetre-sized soft hydrogel beads in an aqueous environment.

Cited by 7 publications

References 36 publications

On a Robust, Sensitive Cell – Free Method for Pseudomonas Sensing and Quantification

On a Robust, Sensitive Cell – Free Method for Pseudomonas Sensing and Quantification

On a Robust, Sensitive Cell-Free Method for Pseudomonas Sensing and Quantification in Microfluidic Templated Hydrogels

Fabricating Shaped and Patterned Supramolecular Multigelator Objects via Diffusion-Adhesion Gel Assembly

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