Reaction-diffusion hydrogels from urease enzyme particles for patterned coatings.

The reaction and diffusion of small molecules is used to initiate the formation of protective polymeric layers, or biofilms, that attach cells to surfaces. Here, inspired by biofilm formation, we present a general method for the growth of hydrogels from urease enzyme-particles by combining production of ammonia with a pH-regulated polymerization reaction in solution. We show through experiments and simulations how the propagating basic front and thiol-acrylate polymerization were continuously maintained by the localized urease reaction in the presence of urea, resulting in hydrogel layers around the enzyme particles at surfaces, interfaces or in motion. The hydrogels adhere the enzyme-particles to surfaces and have a tunable growth rate of the order of 10 µm min-1 that depends on the size and spatial distribution of particles. This approach can be exploited to create enzyme-hydrogels or chemically patterned coatings for applications in biocatalytic flow reactors.

Periodic Nucleation of Calcium Phosphate in a Stirred Biocatalytic Reaction.

Highly ordered superstructures composed of inorganic nanoparticles appear in natural and synthetic systems, however the mechanisms of non-equilibrium self-organization that may be involved are still poorly understood. Herein, we performed a kinetic investigation of the precipitation of calcium phosphate using a process widely found in microorganisms: the hydrolysis of urea by enzyme urease. With high initial ratio of calcium ion to phosphate, periodic precipitation was obtained accompanied by pH oscillations in a well-stirred, closed reactor. We propose that an internal pH-regulated change in the concentration of phosphate ion is the driving force for periodicity. A simple model involving the biocatalytic reaction network coupled with burst nucleation of nanoparticles above a critical supersaturation reproduced key features of the experiments. These findings may provide insight to the self-organization of nanoparticles in biomineralization and improve design strategies of biomaterials for medical applications.

Modelling Bacteria-Inspired Dynamics with Networks of Interacting Chemicals.

One approach to understanding how life-like properties emerge involves building synthetic cellular systems that mimic certain dynamical features of living cells such as bacteria. Here, we developed a model of a reaction network in a cellular system inspired by the ability of bacteria to form a biofilm in response to increasing cell density. Our aim was to determine the role of chemical feedback in the dynamics. The feedback was applied through the enzymatic rate dependence on pH, as pH is an important parameter that controls the rates of processes in cells. We found that a switch in pH can be used to drive base-catalyzed gelation or precipitation of a substance in the external solution. A critical density of cells was required for gelation that was essentially independent of the pH-driven feedback. However, the cell pH reached a higher maximum as a result of the appearance of pH oscillations with feedback. Thus, we conclude that while feedback may not play a vital role in some density-dependent behavior in cellular systems, it nevertheless can be exploited to activate internally regulated cell processes at low cell densities.

Influence of reaction-induced convection on quorum sensing in enzyme-loaded agarose beads.

In theory, groups of enzyme-loaded particles producing an acid or base may show complex behavior including dynamical quorum sensing, the appearance of synchronized oscillations above a critical number or density of particles. Here, experiments were performed with the enzyme urease loaded into mm-sized agarose beads and placed in a solution of urea, resulting in an increase in pH. This behavior was found to be dependent upon the number of beads present in the array; however, reaction-induced convection occurred and plumes of high pH developed that extended to the walls of the reactor. The convection resulted in the motion of the mm-sized particles and conversion of the solution to high pH. Simulations in a simple model of the beads demonstrated the suppression of dynamical quorum sensing in the presence of flow.

Switches induced by quorum sensing in a model of enzyme-loaded microparticles.

Quorum sensing refers to the ability of bacteria and other single-celled organisms to respond to changes in cell density or number with population-wide changes in behaviour. Here, simulations were performed to investigate quorum sensing in groups of diffusively coupled enzyme microparticles using a well-characterized autocatalytic reaction which raises the pH of the medium: hydrolysis of urea by urease. The enzyme urease is found in both plants and microorganisms, and has been widely exploited in engineering processes. We demonstrate how increases in group size can be used to achieve a sigmoidal switch in pH at high enzyme loading, oscillations in pH at intermediate enzyme loading and a bistable, hysteretic switch at low enzyme loading. Thus, quorum sensing can be exploited to obtain different types of response in the same system, depending on the enzyme concentration. The implications for microorganisms in colonies are discussed, and the results could help in the design of synthetic quorum sensing for biotechnology applications such as drug delivery.

Temporal Control of Gelation and Polymerization Fronts Driven by an Autocatalytic Enzyme Reaction.

Chemical systems that remain kinetically dormant until activated have numerous applications in materials science. Herein we present a method for the control of gelation that exploits an inbuilt switch: the increase in pH after an induction period in the urease-catalyzed hydrolysis of urea was used to trigger the base-catalyzed Michael addition of a water-soluble trithiol to a polyethylene glycol diacrylate. The time to gelation (minutes to hours) was either preset through the initial concentrations or the reaction was initiated locally by a base, thus resulting in polymerization fronts that converted the mixture from a liquid into a gel (ca. 0.1 mm min-1). The rate of hydrolytic degradation of the hydrogel depended on the initial concentrations, thus resulting in a gel lifetime of hours to months. In this way, temporal programming of gelation was possible under mild conditions by using the output of an autocatalytic enzyme reaction to drive both the polymerization and subsequent degradation of a hydrogel.

Temporal Control of Gelation and Polymerization Fronts Driven by an Autocatalytic Enzyme Reaction.

Chemical systems that remain kinetically dormant until activated have numerous applications in materials science. Herein we present a method for the control of gelation that exploits an inbuilt switch: the increase in pH after an induction period in the urease-catalyzed hydrolysis of urea was used to trigger the base-catalyzed Michael addition of a water-soluble trithiol to a polyethylene glycol diacrylate. The time to gelation (minutes to hours) was either preset through the initial concentrations or the reaction was initiated locally by a base, thus resulting in polymerization fronts that converted the mixture from a liquid into a gel (ca. 0.1 mm min(-1)). The rate of hydrolytic degradation of the hydrogel depended on the initial concentrations, thus resulting in a gel lifetime of hours to months. In this way, temporal programming of gelation was possible under mild conditions by using the output of an autocatalytic enzyme reaction to drive both the polymerization and subsequent degradation of a hydrogel.

Role of differential transport in an oscillatory enzyme reaction.

As a result of the bell-shaped pH-rate characteristic of enzymatic processes, feedback may arise in enzyme reactions having non-neutral products. This special type of product activation has been shown to lead to self-sustained pH oscillations in an enzyme-loaded membrane. We investigate the possibility of oscillations in a model of the urea-urease reaction, prompted by the recent experimental discovery of feedback in this reaction. An open system is considered in which acid and urea are transported to a cell containing the enzyme. Using linear stability analysis we determine the range of transport coefficients limit cycles may exist for and show that differential transport is required for oscillations in a class of compartmentalized enzyme processes similar to the urea-urease system. We demonstrate that although the transport rate of acid (k(H)) must be greater than that of urea (k(S)) for oscillations in a urease-loaded membrane, bistability is possible for k(S) ≥ k(H).

Motion and interaction of aspirin crystals at aqueous-air interfaces.

Small-molecule amphiphiles such as aspirin have unique properties arising from a combination of an aromatic hydrophobic part and a hydrophilic part. We show that crystals of aspirin are capable of generating convective flows at the air-aqueous interface from both Marangoni effects (through weak surface activity) and capillarity (surface deformations). The flow-driven motion of millimeter-sized crystals was found to depend on the presence of other ions in solution as well as the distance and orientation of the crystals. The interactions lead to the formation of groups of two or more crystals that also underwent motion. The convective flows created by small amphiphile crystals might be exploited in the dynamic self-organization of particles at interfaces.

Two- and three-dimensional standing waves in a reaction-diffusion system.

We observe standing waves of chemical concentration in thin layers [quasi-two-dimensional (2D)] and capillaries [three-dimensional (3D)] containing the aqueous Belousov-Zhabotinsky reaction in a reverse microemulsion stabilized by the ionic surfactant sodium bis-2-ethylhexyl sulfosuccinate (AOT) and with cyclo-octane as the continuous phase. The 3D structures are oscillatory lamellae or square-packed cylinders at high and low volume fractions, respectively, of aqueous droplets. These patterns correspond to oscillatory labyrinthine stripes and square-packed spots in the 2D configuration. Computer simulations, as well as observations in E. coli, give qualitative agreement with the observed patterns and suggest that, in contrast to Turing patterns, the structures are sensitive to the size and shape of the system.

pH wave-front propagation in the urea-urease reaction.

The urease-catalyzed hydrolysis of urea displays feedback that results in a switch from acid (pH ~3) to base (pH ~9) after a controllable period of time (from 10 to >5000 s). Here we show that the spatially distributed reaction can support pH wave fronts propagating with a speed of the order of 0.1-1 mm min(-1). The experimental results were reproduced qualitatively in reaction-diffusion simulations including a Michaelis-Menten expression for the urease reaction with a bell-shaped rate-pH dependence. However, this model fails to predict that at lower enzyme concentrations, the unstirred reaction does not always support fronts when the well-stirred reaction still rapidly switches to high pH.

Turing patterns in the chlorine dioxide-iodine-malonic acid reaction with square spatial periodic forcing.

We use the photosensitive chlorine dioxide-iodine-malonic acid reaction-diffusion system to study wavenumber locking of Turing patterns to two-dimensional "square" spatial forcing, implemented as orthogonal sets of bright bands projected onto the reaction medium. Various resonant structures emerge in a broad range of forcing wavelengths and amplitudes, including square lattices and superlattices, one-dimensional stripe patterns and oblique rectangular patterns. Numerical simulations using a model that incorporates additive two-dimensional spatially periodic forcing reproduce well the experimental observations.

Locking of Turing patterns in the chlorine dioxide-iodine-malonic acid reaction with one-dimensional spatial periodic forcing.

We use the photosensitive chlorine dioxide-iodine-malonic acid reaction-diffusion system to study wavenumber locking of Turing patterns with spatial periodic forcing. Wavenumber-locked stripe patterns are the typical resonant structures that labyrinthine patterns exhibit in response to one-dimensional forcing by illumination when images of stripes are projected on a working medium. Our experimental results reveal that segmented oblique, hexagonal and rectangular patterns can also be obtained. However, these two-dimensional resonant structures only develop in a relatively narrow range of forcing parameters, where the unforced stripe pattern is in close proximity to the domain of hexagonal patterns. Numerical simulations based on a model that incorporates the forcing by illumination using an additive term reproduce well the experimental observations. These findings confirm that additive one-dimensional forcing can generate a two-dimensional resonant response. However, such a response is considerably less robust than the effect of multiplicative forcing.

Tomography of reaction-diffusion microemulsions reveals three-dimensional Turing patterns.

Spatially periodic, temporally stationary patterns that emerge from instability of a homogeneous steady state were proposed by Alan Turing in 1952 as a mechanism for morphogenesis in living systems and have attracted increasing attention in biology, chemistry, and physics. Patterns found to date have been confined to one or two spatial dimensions. We used tomography to study the Belousov-Zhabotinsky reaction in a microemulsion in which the polar reactants are confined to aqueous nanodroplets much smaller than the scale of the stationary patterns. We demonstrate the existence of Turing patterns that can exist only in three dimensions, including curved surfaces, hexagonally packed cylinders, spots, and labyrinthine and lamellar patterns.

Rearrangement dynamics of fishbonelike Turing patterns generated by spatial periodic forcing.

External forcing can greatly affect the evolution of Turing patterns in reaction-diffusion (RD) media. Here, we employ spatially periodic illumination in a photosensitive RD system to create Turing structures that are repetitive in one direction. We then study their relaxation in the absence of light. These unforced, fishbonelike configurations undergo self-reorganization and establish stationary arrangements, some of which fully exhibit, while others partially or completely lose, the symmetry of the initially imposed pattern.

High-frequency oscillations in the Belousov-Zhabotinsky reaction.

Chemical oscillations in the classic Belousov-Zhabotinsky (BZ) system typically have a period of a few minutes, which can be increased significantly by changing the organic substrate. Here we show that by changing the temperature and concentrations, an increase of 3-4 orders of magnitude in the frequency of BZ oscillations can be obtained. At elevated temperatures, in high concentration mixtures, the cerium-catalyzed reaction exhibits sinusoidal oscillations with frequencies of 10 Hz or greater. We report the effect of temperature on the frequency and shape of oscillations in experiments under batch conditions and in a four-variable model. We show that our simple model accurately captures the complex temporal behavior of the system and suggests paths toward even higher frequencies.

Three-dimensional spiral waves in an excitable reaction system: initiation and dynamics of scroll rings and scroll ring pairs.

We report experimental results on spiral and scroll waves in the 1,4-cyclohexanedione Belousov-Zhabotinsky reaction. The propagating concentration waves are detected by two-dimensional photometry and optical tomography. Wave pulses can disappear in front-to-front and front-to-back collisions. This anomaly causes the nucleation of vortices from collisions of three nonrotating waves. In three-dimensional systems, these vortices are scroll rings that rotate around initially circular filaments. Depending on reactant concentrations, the filaments shrink or expand indicating positive and negative filament tensions, respectively. Shrinkage results in vortex annihilation. Expansion is accompanied by filament buckling and bending, which is interpreted as developing Winfree turbulence. We also describe the initiation of scroll ring pairs in four-wave collisions. The two filaments are stacked on top of each other and their motion suggests filament repulsion.

Wave-pinned filaments of scroll waves.

Scroll waves are three-dimensional excitation patterns that rotate around one-dimensional space curves. Typically these filaments are closed loops or end at the system boundary. However, in excitable media with anomalous dispersion, filaments can be pinned to the wake of traveling wave pulses. This pinning is studied in experiments with the 1,4-cyclohexanedione Belousov-Zhabotinsky reaction and a three-variable reaction-diffusion model. We show that wave-pinned filaments are related to the coexistence of rotating and translating wave defects in two dimensions. Filament pinning causes a continuous expansion of the total filament length. It can be ended by annihilating the pinning pulse in a frontal wave collision. Following such an annihilation, the filament connects itself to the system boundary. Its postannihilation shape that is initially the exposed rim of the scroll wave unwinds continuously over numerous rotation periods.

Negative filament tension of scroll rings in an excitable system.

Scroll rings are three-dimensional spiral waves of excitation that rotate around circular filaments. In a modified Belousov-Zhabotinsky reaction, these filaments expand, buckle, and build up gradients in rotation phase. The instability is caused by negative filament tension (-4.3x10;{-4}cm;{2}s) . Initial deformations are strongest in the direction normal to the filament's osculating plane, and their growth rates decrease rapidly with increasing wave number.