Geometric control by active mechanics of epithelial gap closure.

Epithelial wound healing is one of the most important biological processes occurring during the lifetime of an organism. It is a self-repair mechanism closing wounds or gaps within tissues to restore their functional integrity. In this work we derive a new diffuse interface approach for modelling the gap closure by means of a variational principle in the framework of non-equilibrium thermodynamics. We investigate the interplay between the crawling with lamellipodia protrusions and the supracellular tension exerted by the actomyosin cable on the closure dynamics. These active features are modeled as Korteweg forces into a generalised chemical potential. From an asymptotic analysis, we derive a pressure jump across the gap edge in the sharp interface limit. Moreover, the chemical potential diffuses as a Mullins-Sekerka system, and its interfacial value is given by a Gibbs-Thompson relation for its local potential driven by the curvature-dependent purse-string tension. The finite element simulations show an excellent quantitative agreement between the closure dynamics and the morphology of the edge with respect to existing biological experiments. The resulting force patterns are also in good qualitative agreement with existing traction force microscopy measurements. Our results shed light on the geometrical control of the gap closure dynamics resulting from the active forces that are chemically activated around the gap edge.

T cell therapy against cancer: A predictive diffuse-interface mathematical model informed by pre-clinical studies.

T cell therapy has become a new therapeutic opportunity against solid cancers. Predicting T cell behaviour and efficacy would help therapy optimization and clinical implementation. In this work, we model responsiveness of mouse prostate adenocarcinoma to T cell-based therapies. The mathematical model is based on a Cahn-Hilliard diffuse interface description of the tumour, coupled with Keller-Segel type equations describing immune components dynamics. The model is fed by pre-clinical magnetic resonance imaging data describing anatomical features of prostate adenocarcinoma developed in the context of the Transgenic Adenocarcinoma of the Mouse Prostate model. We perform computational simulations based on the finite element method to describe tumor growth dynamics in relation to local T cells concentrations. We report that when we include in the model the possibility to activate tumor-associated vessels and by that increase the number of T cells within the tumor mass, the model predicts higher therapeutic effects (tumor regression) shortly after therapy administration. The simulated results are found in agreement with reported experimental data. Thus, this diffuse-interface mathematical model well predicts T cell behavior in vivo and represents a proof-of-concept for the role such predictive strategies may play in optimization of immunotherapy against cancer.

The Föppl-von Kármán equations of elastic plates with initial stress.

Initially, stressed plates are widely used in modern fabrication techniques, such as additive manufacturing and UV lithography, for their tunable morphology by application of external stimuli. In this work, we propose a formal asymptotic derivation of the Föppl-von Kármán equations for an elastic plate with initial stresses, using the constitutive theory of nonlinear elastic solids with initial stresses under the assumptions of incompressibility and material isotropy. Compared to existing works, our approach allows us to determine the morphological transitions of the elastic plate without prescribing the underlying target metric of the unstressed state of the elastic body. We explicitly solve the derived FvK equations in some physical problems of engineering interest, discussing how the initial stress distribution drives the emergence of spontaneous curvatures within the deformed plate. The proposed mathematical framework can be used to tailor shape on demand, with applications in several engineering fields ranging from soft robotics to four-dimensional printing.

Physical principles of morphogenesis in mushrooms.

Mushroom species display distinctive morphogenetic features. For example, Amanita muscaria and Mycena chlorophos grow in a similar manner, their caps expanding outward quickly and then turning upward. However, only the latter finally develops a central depression in the cap. Here we use a mathematical approach unraveling the interplay between physics and biology driving the emergence of these two different morphologies. The proposed growth elastic model is solved analytically, mapping their shape evolution over time. Even if biological processes in both species make their caps grow turning upward, different physical factors result in different shapes. In fact, we show how for the relatively tall and big A. muscaria a central depression may be incompatible with the physical need to maintain stability against the wind. In contrast, the relatively short and small M. chlorophos is elastically stable with respect to environmental perturbations; thus, it may physically select a central depression to maximize the cap volume and the spore exposure. This work gives fully explicit analytic solutions highlighting the effect of the growth parameters on the morphological evolution, providing useful insights for novel bio-inspired material design.

Modelling cancer cell budding in-vitro as a self-organised, non-equilibrium growth process.

Tissue self-organization into defined and well-controlled three-dimensional structures is essential during development for the generation of organs. A similar, but highly deranged process might also occur during the aberrant growth of cancers, which frequently display a loss of the orderly structures of the tissue of origin, but retain a multicellular organization in the form of spheroids, strands, and buds. The latter structures are often seen when tumors masses switch to an invasive behavior into surrounding tissues. However, the general physical principles governing the self-organized architectures of tumor cell populations remain by and large unclear. In this work, we perform in-vitro experiments to characterize the growth properties of glioblastoma budding emerging from monolayers. We further propose a theoretical model and its finite element implementation to characterize such a topological transition, that is modelled as a self-organised, non-equilibrium phenomenon driven by the trade-off of mechanical forces and physical interactions exerted at cell-cell and cell-substrate adhesions. Notably, the unstable disorder states of uncontrolled cellular proliferation macroscopically emerge as complex spatio-temporal patterns that evolve statistically correlated by a universal law.

Experimental observation of Faraday waves in soft gels.

We report the experimental observation of Faraday waves on soft gels. These were obtained using agarose in a mechanically vibrated cylindrical container. Low driving frequencies induce subharmonic standing waves with spatial structure that conforms to the geometry of the container. We report the experimental observation of the first 15 resonant Faraday wave modes that can be defined by the mode number (n,ℓ) pair. We also characterize the shape of the instability tongue and show the complex dependence upon material properties can be understood as an elastocapillary effect.

Soft Nucleation of an Elastic Crease.

Creasing instability is ubiquitous in soft solids; however, its inception remains enigmatic as it cannot be captured by the standard linearization techniques. It also does not fit the conventional picture of a barrier-crossing nucleation, and instead carries some features of a second order phase transition. Here we show that despite its fundamentally nonlinear nature, creasing has its origin in marginal stability which is, however, obscured by the dominance of long-range elastic interactions. We argue that despite its supercritical (soft) character, creasing bifurcation can be identified by the condition that the (generalized) driving force acting on an incipient stress singularity degenerates. The analytic instability criterion, obtained in this way, shows an excellent agreement with both physical experiments and direct numerical simulations.

On the existence of elastic minimizers for initially stressed materials.

A soft solid is said to be initially stressed if it is subjected to a state of internal stress in its unloaded reference configuration. In physical terms, its stored elastic energy may not vanish in the absence of an elastic deformation, being also dependent on the spatial distribution of the underlying material inhomogeneities. Developing a sound mathematical framework to model initially stressed solids in nonlinear elasticity is key for many applications in engineering and biology. This work investigates the links between the existence of elastic minimizers and the constitutive restrictions for initially stressed materials subjected to finite deformations. In particular, we consider a subclass of constitutive responses in which the strain energy density is taken as a scalar-valued function of both the deformation gradient and the initial stress tensor. The main advantage of this approach is that the initial stress tensor belongs to the group of divergence-free symmetric tensors satisfying the boundary conditions in any given reference configuration. However, it is still unclear which physical restrictions must be imposed for the well-posedness of this elastic problem. Assuming that the constitutive response depends on the choice of the reference configuration only through the initial stress tensor, under given conditions we prove the local existence of a relaxed state given by an implicit tensor function of the initial stress distribution. This tensor function is generally not unique, and can be transformed according to the symmetry group of the material at fixed initial stresses. These results allow one to extend Ball's existence theorem of elastic minimizers for the proposed constitutive choice of initially stressed materials. This article is part of the theme issue 'Rivlin's legacy in continuum mechanics and applied mathematics'.

Matched asymptotic solution for crease nucleation in soft solids.

A soft solid subjected to a large compression develops sharp self-contacting folds at its free surface, known as creases. Creasing is physically different from structural elastic instabilities, like buckling or wrinkling. Indeed, it is a fully nonlinear material instability, similar to a phase-transformation. This work provides theoretical insights of the physics behind crease nucleation. Creasing is proved to occur after a global bifurcation allowing the co-existence of an outer deformation and an inner solution with localised self-contact at the free surface. The most fundamental result here is the analytic prediction of the nucleation threshold, in excellent agreement with experiments and numerical simulations. A matched asymptotic solution is given within the intermediate region between the two co-existing states. The self-contact acts like the point-wise disturbance in the Oseen's correction for the Stokes flow past a circle. Analytic expressions of the matching solution and its range of validity are also derived.

Solid tumors are poroelastic solids with a chemo-mechanical feedback on growth.

The experimental evidence that a feedback exists between growth and stress in tumors poses challenging questions. First, the rheological properties (the "constitutive equations") of aggregates of malignant cells are still a matter of debate. Secondly, the feedback law (the "growth law") that relates stress and mitotic-apoptotic rate is far to be identified. We address these questions on the basis of a theoretical analysis of in vitro and in vivo experiments that involve the growth of tumor spheroids. We show that solid tumors exhibit several mechanical features of a poroelastic material, where the cellular component behaves like an elastic solid. When the solid component of the spheroid is loaded at the boundary, the cellular aggregate grows up to an asymptotic volume that depends on the exerted compression. Residual stress shows up when solid tumors are radially cut, highlighting a peculiar tensional pattern. By a novel numerical approach we correlate the measured opening angle and the underlying residual stress in a sphere. The features of the mechanobiological system can be explained in terms of a feedback of mechanics on the cell proliferation rate as modulated by the availability of nutrient, that is radially damped by the balance between diffusion and consumption. The volumetric growth profiles and the pattern of residual stress can be theoretically reproduced assuming a dependence of the target stress on the concentration of nutrient which is specific of the malignant tissue.

Rayleigh-Taylor instability in soft elastic layers.

This work investigates the morphological stability of a soft body composed of two heavy elastic layers attached to a rigid surface and subjected only to the bulk gravity force. Using theoretical and computational tools, we characterize the selection of different patterns as well as their nonlinear evolution, unveiling the interplay between elastic and geometric effects for their formation. Unlike similar gravity-induced shape transitions in fluids, such as the Rayleigh-Taylor instability, we prove that the nonlinear elastic effects saturate the dynamic instability of the bifurcated solutions, displaying a rich morphological diagram where both digitations and stable wrinkling can emerge. The results of this work provide important guidelines for the design of novel soft systems with tunable shapes, with several applications in engineering sciences.This article is part of the themed issue 'Patterning through instabilities in complex media: theory and applications.'

Magnetic domain wall tweezers: a new tool for mechanobiology studies on individual target cells.

In vitro tests are of fundamental importance for investigating cell mechanisms in response to mechanical stimuli or the impact of the genotype on cell mechanical properties. In particular, the application of controlled forces to activate specific bio-pathways and investigate their effects, mimicking the role of the cellular environment, is becoming a prominent approach in the emerging field of mechanobiology. Here, we present an on-chip device based on magnetic domain wall manipulators, which allows the application of finely controlled and localized forces on target living cells. In particular, we demonstrate the application of a magnetic force in the order of hundreds of pN on the membrane of HeLa cells cultured on-chip, via manipulation of 1 µm superparamagnetic beads. Such a mechanical stimulus produces a sizable local indentation of the cellular membrane of about 2 µm. Upon evaluation of the beads' position within the magnetic field originated by the domain wall, the force applied during the experiments is accurately quantified via micromagnetic simulations. The obtained value is in good agreement with that calculated by the application of an elastic model to the cellular membrane.

On residual stresses and homeostasis: an elastic theory of functional adaptation in living matter.

Living matter can functionally adapt to external physical factors by developing internal tensions, easily revealed by cutting experiments. Nonetheless, residual stresses intrinsically have a complex spatial distribution, and destructive techniques cannot be used to identify a natural stress-free configuration. This work proposes a novel elastic theory of pre-stressed materials. Imposing physical compatibility and symmetry arguments, we define a new class of free energies explicitly depending on the internal stresses. This theory is finally applied to the study of arterial remodelling, proving its potential for the non-destructive determination of the residual tensions within biological materials.

Finite element simulations of the active stress in the imaginal disc of the Drosophila Melanogaster.

During the larval stages of development, the imaginal disc of Drosphila Melanogaster is composed by a monolayer of epithelial cells, which undergo a strain actively produced by the cells themselves. The well-organized collective contraction produces a stress field that seemingly has a double morphogenetic role: it orchestrates the cellular organization towards the macroscopic shape emergence while simultaneously providing a local information on the organ size. Here we perform numerical simulations of such a mechanical control on morphogenesis at a continuum level, using a three-dimensional finite model that accounts for the active cell contraction. The numerical model is able to reproduce the (few) known qualitative characteristics of the tensional patterns within the imaginal disc of the fruit fly. The computed stress components slightly deviate from planarity, thus confirming the previous theoretical assumptions of a nonlinear elastic analytical model, and enforcing the hypothesis that the spatial variation of the mechanical stress may act as a size regulating signal that locally scales with the global dimension of the domain.

Elastocapillarity can control the formation and the morphology of beads-on-string structures in solid fibers.

Beads-on-string patterns have been experimentally observed in solid cylinders for a wide range of material properties and structural lengths, from millimetric soft gels to nanometric hard fibers. In this work, we combine theoretical analysis and numerical tools to investigate the formation and nonlinear dynamics of such beaded structures. We show that this morphological transition is driven by elastocapillarity, i.e., a complex interplay between the effects of surface tension and bulk elasticity. Unlike buckling or wrinkling, the presence of an axial elongation is found here to favor the onset of fiber beading, in agreement with existing experimental results on electrospun fibers, hydrogels, and nerves. Our results also prove that the applied stretch can be used in fabrication techniques to control the morphology of the emerging beads-on-string patterns. Such quantitative predictions open the way for several applications, from tissue engineering to the design of stretchable electronics and the microfabrication of functionalized surfaces.

Pattern selection in growing tubular tissues.

Tubular organs display a wide variety of surface morphologies including circumferential and longitudinal folds, square and hexagonal undulations, and finger-type protrusions. Surface morphology is closely correlated to tissue function and serves as a clinical indicator for physiological and pathological conditions, but the regulators of surface morphology remain poorly understood. Here, we explore the role of geometry and elasticity on the formation of surface patterns. We establish morphological phase diagrams for patterns selection and show that increasing the thickness or stiffness ratio between the outer and inner tubular layers induces a gradual transition from circumferential to longitudinal folding. Our results suggest that physical forces act as regulators during organogenesis and give rise to the characteristic circular folds in the esophagus, the longitudinal folds in the valves of Kerckring, the surface networks in villi, and the crypts in the large intestine.

Mechano-transduction in tumour growth modelling.

The evolution of biological systems is strongly influenced by physical factors, such as applied forces, geometry or the stiffness of the micro-environment. Mechanical changes are particularly important in solid tumour development, as altered stromal-epithelial interactions can provoke a persistent increase in cytoskeletal tension, driving the gene expression of a malignant phenotype. In this work, we propose a novel multi-scale treatment of mechano-transduction in cancer growth. The avascular tumour is modelled as an expanding elastic spheroid, whilst growth may occur both as a volume increase and as a mass production within a cell rim. Considering the physical constraints of an outer healthy tissue, we derive the thermo-dynamical requirements for coupling growth rate, solid stress and diffusing biomolecules inside a heterogeneous tumour. The theoretical predictions successfully reproduce the stress-dependent growth curves observed by in vitro experiments on multicellular spheroids.

Morpho-elasticity of intestinal villi.

Villi are ubiquitous structures in the intestine of all vertebrates, originating from the embryonic development of the epithelial mucosa. Their morphogenesis has similar stages in living organisms but different forming mechanisms. In this work, we model the emergence of the bi-dimensional undulated patterns in the intestinal mucosa from which villi start to elongate. The embryonic mucosa is modelled as a growing thick-walled cylinder, and its mechanical behaviour is described using an hyperelastic constitutive model, which also accounts for the anisotropic characteristics of the reinforcing fibres at the microstructural level. The occurrence of surface undulations is investigated using a linear stability analysis based on the theory of incremental deformations superimposed on a finite deformation. The Stroh formulation of the incremental boundary value problem is derived, and a numerical solution procedure is implemented for calculating the growth thresholds of instability. The numerical results are finally discussed with respect to different growth and materials properties. In conclusion, we demonstrate that the emergence of intestinal villi in embryos is triggered by a differential growth between the mucosa and the mesenchymal tissues. The proposed model quantifies how both the geometrical and the mechanical properties of the mucosa drive the formation of previllous structures in embryos.

Surface instability of a gel disc in swelling.

The swelling of a soft disc made of polymeric gel and attached to a fixed substrate is modeled using a variational method in nonlinear elasticity. A linear stability analysis is performed to detect the onset of a surface instability. An exact solution of the perturbed disc is found, and both the threshold values of the growth rates and the surface morphology are derived analytically.

Buckling instability in growing tumor spheroids.

A growing tumor is subjected to intrinsic physical forces, arising from the cellular turnover in a spatially constrained environment. This work demonstrates that such residual solid stresses can provoke a buckling instability in heterogeneous tumor spheroids. The growth rate ratio between the outer shell of proliferative cells and the inner necrotic core is the control parameter of this instability. The buckled morphology is found to depend both on the elastic and the geometric properties of the tumor components, suggesting a key role of residual stresses for promoting tumor invasiveness.