Cellular sensing of micron-scale curvature: a frontier in understanding the microenvironment.

The vast majority of cell biological studies examine function and molecular mechanisms using cells on flat surfaces: glass, plastic and more recently elastomeric polymers. While these studies have provided a wealth of valuable insight, they fail to consider that most biologically occurring surfaces are curved, with a radius of curvature roughly corresponding to the length scale of cells themselves. Here, we review recent studies showing that cells detect and respond to these curvature cues by adjusting and re-orienting their cell bodies, actin fibres and nuclei as well as by changing their transcriptional programme. Modelling substratum curvature has the potential to provide fundamental new insight into cell behaviour and function in vivo.

Edges impose planar alignment in nematic monolayers by directing cell elongation and enhancing migration.

Boundaries play an important role in the emergence of nematic order in classical liquid crystal systems; we explore their importance in adhesive cells that form active nematics. In particular, we study how cells are affected by an edge, which in our experiments is a boundary between adhesive and non-adhesive domains on a planar surface. We find that such edges induce elongation and direct the migration of isolated fibroblasts. In confluent monolayers, these elongated cells co-align and migrate to form an active, two-dimensional nematic structure in which edges enforce planar alignment and provide local slip to streams of cells that move along them. On an adhesive square island of dimensions 1 mm × 1 mm, cells near the edges in confluent nematic monolayers have enhanced alignment and velocity. The corners of the adhesive island seed defects with signs that depend on the direction of the motion of the streams of cells that meet there. Distortions emerge with rotations of -π/2 to form a -1/4 defect for streams that move clockwise or counterclockwise, and +π/2 to form a +1/4 defect for converging streams. We explore how cells transmit alignment information to each other in the absence of an edge by studying cell pairs and find that while such pairs do co-align, this alignment is only transient and short lived. These results shed light on the importance of edges in imposing nematic order in confluent monolayers and how edges can be used as tools to pattern the long-range organization of cells for tissue engineering applications.

Gaussian Curvature Directs Stress Fiber Orientation and Cell Migration.

We show that substrates with nonzero Gaussian curvature influence the organization of stress fibers and direct the migration of cells. To study the role of Gaussian curvature, we developed a sphere-with-skirt surface in which a positive Gaussian curvature spherical cap is seamlessly surrounded by a negative Gaussian curvature draping skirt, both with principal radii similar to cell-length scales. We find significant reconfiguration of two subpopulations of stress fibers when fibroblasts are exposed to these curvatures. Apical stress fibers in cells on skirts align in the radial direction and avoid bending by forming chords across the concave gap, whereas basal stress fibers bend along the convex direction. Cell migration is also strongly influenced by the Gaussian curvature. Real-time imaging shows that cells migrating on skirts repolarize to establish a leading edge in the azimuthal direction. Thereafter, they migrate in that direction. This behavior is notably different from migration on planar surfaces, in which cells typically migrate in the same direction as the apical stress fiber orientation. Thus, this platform reveals that nonzero Gaussian curvature not only affects the positioning of cells and alignment of stress fiber subpopulations but also directs migration in a manner fundamentally distinct from that of migration on planar surfaces.

Curvature and Rho activation differentially control the alignment of cells and stress fibers.

In vivo, cells respond to a host of physical cues ranging from substrate stiffness to the organization of micro- and nanoscale fibrous networks. We show that macroscale substrates with radii of curvature from tens to hundreds of micrometers influence cell alignment. In a model system of fibroblasts, isolated cells aligned strongly in the axial direction on cylinders with radii similar to the cell length and more weakly on cylinders of much larger radius. Isolated vascular smooth muscle cells did not align as effectively as fibroblasts. However, both cell types aligned robustly in weak curvature fields when in confluent monolayers. We identified two distinct populations of stress fibers in both cell types: long, apical stress fibers that aligned axially and short, basal stress fibers that aligned circumferentially. Circumferential alignment of the basal stress fibers is in apparent disagreement with a long-standing hypothesis that energetic penalties for bending enforce axial alignment on cylinders. To explore this phenomenon, we manipulated stress fibers by activating Rho, a small guanosine triphosphatase that regulates stress fiber assembly. In response, apical stress fibers disassembled, whereas basal stress fibers thickened and aligned more strongly in the circumferential direction. By activating Rho in confluent monolayers of vascular smooth muscle cells, we recapitulated the circumferential alignment pattern of F-actin within these cells that is observed in cylindrical vessels in vivo. In agreement with recent theory, these results suggest that stress fiber bending penalties are overcome when stress fiber contractility is enhanced and motivate deeper study of the mechanics of these distinct stress fiber populations.

Rough Adhesive Hydrogels (RAd gels) for Underwater Adhesion.

In this work, underwater adhesion is achieved between biocompatible hydrogels and a suite of substrates. Surface roughness, which is typically detrimental for adhesion in air, is shown to be beneficial for underwater adhesion. Contact between the hydrogels with macroscopically flat substrates, and the resulting nonspecific chemical interaction, is facilitated by surface roughness, which enables drainage of the lubricating fluid layer. Hydrogel composition plays an important role in tuning the gel elasticity and interaction with the substrate. Hydrogels that are adhesive on two sides are synthesized for potential use as versatile adhesives in various applications.

Smectic Gardening on Curved Landscapes.

Focal conic domains (FCDs) form in smectic-A liquid crystal films with hybrid anchoring conditions with eccentricity and size distribution that depend strongly on interface curvature. Assemblies of FCDs can be exploited in settings ranging from optics to material assembly. Here, using micropost arrays with different shapes and arrangement, we assemble arrays of smectic flower patterns, revealing their internal structure as well as defect size, location, and distribution as a function of interface curvature, by imposing positive, negative, or zero Gaussian curvature at the free surface. We characterize these structures, relating free surface topography, substrate anchoring strength, and FCD distribution. Whereas the largest FCDs are located in the thickest regions of the films, the distribution of sizes is not trivially related to height, due to Apollonian tiling. Finally, we mold FCDs around microposts of complex shape and find that FCD arrangements are perturbed near the posts, but are qualitatively similar far from the posts where the details of the confining walls and associated curvature fields decay. This ability to mold FCD defects into a variety of hierarchical assemblies by manipulating the interface curvature paves the way to create new optical devices, such as compound eyes, via a directed assembly scheme.

The (dys)functional extracellular matrix.

The extracellular matrix (ECM) is a major component of the biomechanical environment with which cells interact, and it plays important roles in both normal development and disease progression. Mechanical and biochemical factors alter the biomechanical properties of tissues by driving cellular remodeling of the ECM. This review provides an overview of the structural, compositional, and mechanical properties of the ECM that instruct cell behaviors. Case studies are reviewed that highlight mechanotransduction in the context of two distinct tissues: tendons and the heart. Although these two tissues demonstrate differences in relative cell-ECM composition and mechanical environment, they share similar mechanisms underlying ECM dysfunction and cell mechanotransduction. Together, these topics provide a framework for a fundamental understanding of the ECM and how it may vary across normal and diseased tissues in response to mechanical and biochemical cues. This article is part of a Special Issue entitled: Mechanobiology.