Geometric confinement guides topological defect pairings and emergent flow in nematic cell populations.

Topological defects in nematically aligned cell populations play a critical role in modulating collective motion, ranging from microbial colonies to epithelial tissues. Despite the potential of manipulating such topological defects to control diverse self-organized structures and collective dynamics, controlling the position of defects in active matter remains a challenging area of research. In this study, we investigated the geometry-guided control of defect positioning and alignment in a nematic cell population by imposing spatial constraints consisting of two or three overlapping circular boundaries. The confined cell population exhibited a paired and ordered distribution of half-integer topological defects that remained stable even when the size of the spatial constraint was altered using geometric parameters. These defects direct the inward flow of cells, induced by the curved boundary shape, towards the geometric center of the confined space. This inward flow contributes to an increase in a local cell density, and furthermore the geometry-induced nematic order provides mechanical stimulation to confined cells, as indicated by the elongated cell nucleus. Our geometry-based approach sets the foundation for controlling defect pairing and provides insights into the interplay among geometry, topology, and collective dynamics.

Collective motion of epithelial cells along a wrinkled 3D-buckled hydrogel.

Epithelial cells migrate autonomously by aligning and inducing a collective motion. Controlling the collective motion of epithelial cells in geometrically confined environments is important for understanding physiological processes such as wound healing and self-organized morphogenesis. However, collective migration under a three-dimensional (3D) curved surface resembling living epithelial tissue has not yet been explored. In this study, we investigated the collective motion of a 3D-buckled polyacrylamide (PAAm) gel that mimics the shape of folds and wrinkles of epithelial tissue to understand the geometric effects of collective motion. We found that the velocity correlation in the space near the hydrogel boundary showed a periodic change that correlated with the wrinkled folding of the hydrogel pattern. Furthermore, the characteristic length of the velocity correlation increased proportionally with the wavelength of wrinkled folding. These observations indicated that the hydrogel pattern could steer the collective motion of epithelial cells over long distances. Our study also suggests that the wrinkled design of the hydrogel is a versatile platform for studying the geometric effect of a curved surface on complex epithelial cell dynamics.

Exploring order in active turbulence: Geometric rule and pairing order transition in confined bacterial vortices.

Ordered collective motion emerges in a group of autonomously motile elements (known as active matter) as their density increases. Microswimmers, such as swimming bacteria, have been extensively studied in physics and biology. A dense suspension of bacteria forms seemingly chaotic turbulence in viscous fluids. Interestingly, this active turbulence driven by bacteria can form a hidden ensemble of many vortices. Understanding the active turbulence in a bacterial suspension can provide physical principles for pattern formation and insight into the instability underlying biological phenomena. This review presents recent findings regarding ordered structures causing active turbulence and discusses a physical approach for controlling active turbulence via geometric confinement. When the active matter is confined in a compartment with a size comparable to the correlation length of the collective motion, vortex-like rotation appears, and the vortex pairing order is indicated by the patterns of interacting vortices. Additionally, we outline the design principle for controlling collective motions via the geometric rule of the vortex pairing, which may advance engineering microdevices driven by a group of active matter. This article is an extended version of the Japanese article, Ordered Structure and Geometric Control of Active Matter in Dense Bacterial Suspensions, published in SEIBUTSU BUTSURI Vol. 60, p. 13-18 (2020).

Controlling Collective Motion of Kinesin-Driven Microtubules via Patterning of Topographic Landscapes.

Biomolecular motor proteins that generate forces by consuming chemical energy obtained from ATP hydrolysis play pivotal roles in organizing cytoskeletal structures in living cells. An ability to control cytoskeletal structures would benefit programmable protein patterning; however, our current knowledge is limited because of the underdevelopment of engineering approaches for controlling pattern formation. Here, we demonstrate the controlling of self-assembled patterns of microtubules (MTs) driven by kinesin motors by designing the boundary shape in fabricated microwells. By manipulating the collision angle of gliding MTs defined by the boundary shape, the self-assembly of MTs can be controlled to form protruding bundle and bridge patterns. Corroborated by the theory of self-propelled rods, we further show that the alignment of MTs determines the transition between the assembled patterns, providing a blueprint to reconstruct bridge structures in microchannels. Our findings introduce the tailoring of the self-organization of cytoskeletons and motor proteins for nanotechnological applications.

Edge current and pairing order transition in chiral bacterial vortices.

Bacterial suspensions show turbulence-like spatiotemporal dynamics and vortices moving irregularly inside the suspensions. Understanding these ordered vortices is an ongoing challenge in active matter physics, and their application to the control of autonomous material transport will provide significant development in microfluidics. Despite the extensive studies, one of the key aspects of bacterial propulsion has remained elusive: The motion of bacteria is chiral, i.e., it breaks mirror symmetry. Therefore, the mechanism of control of macroscopic active turbulence by microscopic chirality is still poorly understood. Here, we report the selective stabilization of chiral rotational direction of bacterial vortices in achiral circular microwells sealed by an oil/water interface. The intrinsic chirality of bacterial swimming near the top and bottom interfaces generates chiral collective motions of bacteria at the lateral boundary of the microwell that are opposite in directions. These edge currents grow stronger as bacterial density increases, and, within different top and bottom interfaces, their competition leads to a global rotation of the bacterial suspension in a favored direction, breaking the mirror symmetry of the system. We further demonstrate that chiral edge current favors corotational configurations of interacting vortices, enhancing their ordering. The intrinsic chirality of bacteria is a key feature of the pairing order transition from active turbulence, and the geometric rule of pairing order transition may shed light on the strategy for designing chiral active matter.

Geometric Effect for Biological Reactors and Biological Fluids.

As expressed "God made the bulk; the surface was invented by the devil" by W. Pauli, the surface has remarkable properties because broken symmetry in surface alters the material properties. In biological systems, the smallest functional and structural unit, which has a functional bulk space enclosed by a thin interface, is a cell. Cells contain inner cytosolic soup in which genetic information stored in DNA can be expressed through transcription (TX) and translation (TL). The exploration of cell-sized confinement has been recently investigated by using micron-scale droplets and microfluidic devices. In the first part of this review article, we describe recent developments of cell-free bioreactors where bacterial TX-TL machinery and DNA are encapsulated in these cell-sized compartments. Since synthetic biology and microfluidics meet toward the bottom-up assembly of cell-free bioreactors, the interplay between cellular geometry and TX-TL advances better control of biological structure and dynamics in vitro system. Furthermore, biological systems that show self-organization in confined space are not limited to a single cell, but are also involved in the collective behavior of motile cells, named active matter. In the second part, we describe recent studies where collectively ordered patterns of active matter, from bacterial suspensions to active cytoskeleton, are self-organized. Since geometry and topology are vital concepts to understand the ordered phase of active matter, a microfluidic device with designed compartments allows one to explore geometric principles behind self-organization across the molecular scale to cellular scale. Finally, we discuss the future perspectives of a microfluidic approach to explore the further understanding of biological systems from geometric and topological aspects.

Geometry-driven collective ordering of bacterial vortices.

Controlling the phases of matter is a challenge that spans from condensed materials to biological systems. Here, by imposing a geometric boundary condition, we study the controlled collective motion of Escherichia coli bacteria. A circular microwell isolates a rectified vortex from disordered vortices masked in the bulk. For a doublet of microwells, two vortices emerge but their spinning directions show transition from parallel to anti-parallel. A Vicsek-like model for confined self-propelled particles gives the point where the two spinning patterns occur in equal probability and one geometric quantity governs the transition as seen in experiments. This mechanism shapes rich patterns including chiral configurations in a quadruplet of microwells, thus revealing a design principle of active vortices.