Animal morphogenesis involves complex tissue deformation processes, which require tight control over tissue rheology. Yet, it remains insufficiently understood how tissue rheology results from the interplay between cellular packing and forces, such as cortical tension or cell-cell adhesion. We follow a biomimetic approach to study this interplay, using oil droplets with tunable adhesion strength to mimic adhesive cells. We expose emulsions to cyclic shear and use a geometric method to quantify their rheology using only imaging data. We find that emulsions made of two droplet types change yielding behavior across subsequent shear cycles. Combining this with vertex model simulations, we show that this shift is due to a progressive compaction, which only occurs with a high adhesion differential and only under oscillatory shear. Our work thus demonstrates how gradients observed during development can lead to gradients in tissue rheology. Moreover, progressive compaction suggests the emergence of a pumping mechanism, which potentially acts in many cellular materials, from foams to tissues.

Mechanical contributions are crucial regulators of diverse biological processes, yet their in vivo measurement remains challenging due to limitations of current techniques that can be destructive or require complex dedicated setups. This study introduces a novel method to synthesize biocompatible, self-functionalizing stress sensors based on inverted emulsions that can be used to probe stresses inside tissues but can also locally perturb the biological environment through specific binder presentation or drug delivery. We engineered an optimal design for these inverted emulsions, focusing on finding the balance between the two contradictory constraints: achieving low surface tension for deformability while maintaining emulsion instability for efficient self-functionalization and drug release. Proof-of-concept experiments in both agarose gels and complex biological systems, including brain organoids and zebrafish embryos, confirm the droplets ability to deform in response to mechanical stress applied within the tissue, to self-functionalize and to release encapsulated molecules locally. These versatile sensors offer a method for noninvasive stress measurements and targeted chemical delivery within living biological tissues, giving the potential to overcome current technical barriers in biophysical studies. Left 3 panels: Inverted emulsion droplets self-functionalize over time once injected in a biological medium. Right panel: After self-functionalization, the deformability of the droplets allows for the measurment of local stresses.

In tissues, cells in direct physical contact with each other can exchange ions or molecules via protein clusters called gap junctions, that form channels across the membranes of adjacent cells. Here, we use a simplified biomimetic approach, coupled with theoretical modeling, to unravel the physical mechanisms controlling such transport. Tissues are mimicked with 2D hexagonal networks of monodisperse aqueous droplets connected by lipid membranes called droplet interface bilayers (DIBs) decorated with α-hemolysin (αHL) transmembrane proteins forming nanopores through heptamerization in the membrane. The diffusion of calcein across 2D DIB networks is thoroughly studied using epifluorescence microscopy at various αHL concentrations. The results are successfully reproduced with a continuous time random walk model in hexagonal networks with an average waiting time increasing nonlinearly with the concentration of pore monomers.

We mimic the mechanical response of touch mechanoreceptors by that of a gas cavity embedded in an elastic semi-cylinder, as a fingertip analogue. Using tribological experiments combined with optical imaging, we measure the dynamics and deformation of the cavity as the semi-cylinder is put in static contact or slid against model rough surfaces at constant normal force and velocity. We propose an elastic model to predict the cavity deformation under normal load showing that membrane mechanical stresses are anisotropic and we discuss its possible biological consequences. In friction experiments, we show that the cavity shape fluctuations allow for texture discriminations.

We present a simple method to produce giant lipid pseudo-vesicles (vesicles with an oily cap on the top), trapped in an agarose gel. The method can be implemented using only a regular micropipette and relies on the formation of a water/oil/water double droplet in liquid agarose. We characterize the produced vesicle with fluorescence imaging and establish the presence and integrity of the lipid bilayer by the successful insertion of [Formula: see text]-Hemolysin transmembrane proteins. Finally, we show that the vesicle can be easily mechanically deformed, non-intrusively, by indenting the surface of the gel.

We present a simple method to produce giant lipid pseudo-vesicles (vesicles with an oily cap on the top), trapped in an agarose gel. The method can be implemented using only a regular micropipette and relies on the formation of a water/oil/water double droplet in liquid agarose. We characterize the produced vesicle with fluorescence imaging and establish the presence and integrity of the lipid bilayer by the successful insertion of α-Hemolysin transmembrane proteins. Finally, we show that the vesicle can be easily mechanically deformed, non-intrusively, by indenting the surface of the gel.

Tissues are subjected to large external forces and undergo global deformations during morphogenesis. We use synthetic analogues of tissues to study the impact of cell-cell adhesion on the response of cohesive cellular assemblies under such stresses. In particular, we use biomimetic emulsions in which the droplets are functionalized in order to exhibit specific droplet-droplet adhesion. We flow these emulsions in microfluidic constrictions and study their response to this forced deformation via confocal microscopy. We find that the distributions of avalanche sizes are conserved between repulsive and adhesive droplets. However, adhesion locally impairs the rupture of droplet-droplet contacts, which in turn pulls on the rearranging droplets. As a result, adhesive droplets are a lot more deformed along the axis of elongation in the constriction. This finding could shed light on the origin of polarization processes during morphogenesis.

To mimic the mechanical response of passive biological cilia in complex fluids, we study the bending dynamics of an anchored elastic fiber submitted to a dilute granular suspension under shear. We show that the bending fluctuations of the fiber accurately encode minute variations of the granular suspension concentration. Indeed, besides the stationary bending induced by the continuous phase flow, the passage of each single particle induces an additional deflection. We demonstrate that the dominant particle/fiber interaction arises from contacts of the particles with the fiber, and we propose a simple elastohydrodynamics model to predict their amplitude. Our results provide a mechanistic and statistical framework to describe particle detection by biological ciliated systems.

Characterizing the mechanical properties of tissues is key for the understanding of fundamental biological processes such as morphogenesis or tumor progression. In particular, the intercellular adhesion forces, mediated by transmembrane proteins like cadherins, are expected to control the topology and viscoelastic behavior of tissues under mechanical stress. In order to understand the influence of adhesion in tissues, we use biomimetic emulsions in which droplets mimic cells and adhere to each other through specific bonds. Here, we tune both the binding energy of the adhesive inter-droplets contacts as well as the fraction of contacts that are adhesive, thereby defining a so-called adhesiveness. Our experimental results show that adhesion prevents the emergence of local order in emulsions even at high packing fractions by preventing energetically costly droplet rearrangements. By studying the deformation of droplets within packings with different average adhesiveness values, we reveal the existence of a threshold value of adhesiveness above which all droplets in a packing are deformed as adhesive ones irrespective of their local adhesive properties. We show that this critical adhesiveness coincides with the threshold for percolation of adhesive structures throughout the tissue. From a biological point of view, this indicates that only a fraction of adhesive cells would be sufficient to tune the global mechanical properties of a tissue, which would be critical during processes such as morphogenesis.

Many living systems use assemblies of soft and slender structures whose deflections allow them to mechanically probe their immediate environment. In this work, we study the collective response of artificial soft hair assemblies to a shear flow by imaging their deflections. At all hair densities, the deflection is found to be proportional to the local shear stress with a proportionality factor that decreases with density. The measured collective stiffening of hairs is modeled both with a microscopic elastohydrodynamic model that takes into account long-range hydrodynamic hair-hair interactions and a phenomenological model that treats the hair assemblies as an effective porous medium. While the microscopic model is in reasonable agreement with the experiments at low hair density, the phenomenological model is found to be predictive across the entire density range.

To mimic the mechanical response of passive biological cilia in complex fluids, we study the bending dynamics of an anchored elastic fiber submitted to a dilute granular suspension under shear. We show that the bending fluctuations of the fiber accurately encode minute variations of the granular suspension concentration. Indeed, besides the stationary bending induced by the continuous phase flow, the passage of each single particle induces an additional deflection. We demonstrate that the dominant particle/fiber interaction arises from direct contacts of the particles with the fiber and we propose a simple elastohydrodynamics model to predict their amplitude. Our results shed light on the extreme mechanical sensitivity of biological cilia to detect the presence of solid particles in their vicinity and bring a physical framework to describe their dynamics in particulate flows.

We study the elasto-plastic behavior of dense attractive emulsions under a mechanical perturbation. The attraction is introduced through non-specific depletion interactions between the droplets and is controlled by changing the concentration of surfactant micelles in the continuous phase. We find that such attractive forces are not sufficient to induce any measurable modification on the scalings between the local packing fraction and the deformation of the droplets. However, when the emulsions are flowed through 2D microfluidic constrictions, we uncover a measurable effect of attraction on their elasto-plastic response. Indeed, we measure higher levels of deformation inside the constriction for attractive droplets. In addition, we show that these measurements correlate with droplet rearrangements that are spatially delayed in the constriction for higher attraction forces.

A biomimetic model of cell-cell communication was developed to probe the passive molecular transport across ion channels inserted in synthetic lipid bilayers formed between contacting droplets arranged in a linear array. Diffusion of a fluorescent probe across the array was measured for different pore concentrations. The diffusion characteristic timescale is found to vary nonlinearly with the pore concentration. Our measurements are successfully modeled by a continuous time random walk description whose waiting time is the first exit time from a droplet through a cluster of pores. The size of the cluster of pores is found to increase with their concentration. Our results provide a direct link between the mesoscopic permeation properties and the microscopic characteristics of the pores, such as their number, size, and spatial arrangement.

An experimental biomimetic tongue-palate system has been developed to probe human in-mouth texture perception. Model tongues are made from soft elastomers patterned with fibrillar structures analogous to human filiform papillae. The palate is represented by a rigid flat plate parallel to the plane of the tongue. To probe the behaviour under physiological flow conditions, deflections of model papillae are measured using a novel fluorescent imaging technique enabling sub-micrometre resolution of the displacements. Using optically transparent Newtonian liquids under steady shear flow, we show that deformations of the papillae allow their viscosity to be determined from 1 Pa s down to the viscosity of water (1 mPa s), in full quantitative agreement with a previously proposed model (Lauga et al. 2016 Front. Phys.4, 35 (doi:10.3389/fphy.2016.00035)). The technique is further validated for a shear-thinning and optically opaque dairy system.

We study the elasto-plastic behavior of dense attractive emulsions under mechanical perturbation. The attraction is introduced through non-specific depletion interactions between the droplets and is controlled by changing the concentration of surfactant micelles in the continuous phase. We find that such attractive forces are not sufficient to induce any measurable modification on the scalings between the local packing fraction and the deformation of the droplets. However, when the emulsions are flown through 2D microfluidic constrictions, we uncover a measurable effect of attraction on their elasto-plastic response. Indeed, we measure higher levels of deformation inside the constriction for attractive droplets. In addition, we show that these measurements correlate with droplet rearrangements that are spatially delayed in the constriction for higher attraction forces.

A biomimetic model of cell-cell communication was developed to probe the passive molecular transport across ion channels inserted in synthetic lipid bilayers formed between contacting droplets arranged in a linear array. Diffusion of a fluorescent probe across the array was measured for different pore concentrations. The diffusion characteristic time scale is found to vary non-linearly with the pore concentration. Our measurements are successfully modeled by a continuous time random walk description, whose waiting time is the first exit time from a droplet through a cluster of pores. The size of the cluster of pores is found to increase with their concentration. Our results provide a direct link between the mesoscopic permeation properties and the microscopic characteristics of the pores such as their number, size and spatial arrangement.

Émulsions biomimétiques pour sonder la mécanique des tissus J’utilise une approche biomimétique pour comprendre les mécanismes physiques qui sous-tendent le remodelage des tissus biologiques. J'étudie en particulier le rôle des interactions entre adhésion et forces mécaniques dans la morphogénèse, processus lors duquel les tissus sont soumis à des élongations et repliements successifs qui modèlent la forme des embryons. Ce sont ces aspects mécaniques que nous étudions par deux approches complémentaires in vitro et in vivo dans un cadre simplifié. En particulier, nous utilisons des émulsions biomimétiques constituées dont les propriétés mécaniques et adhésives miment celles des cellules d’un tissu. Ces émulsions biomimétiques sont stablisées par des phospholipides et peuvent être fonctionnalisées par des ligands spécifiques. Le premier aspect de mon projet consiste à étudier in vitro le comportement mécanique de d'émulsions adhésives soumises à des perturbations mécaniques, en les injectant dans des constrictions microfluidiques de géométrie contrôlée. L'adhesion entre les gouttes est créée soit par des forces de déplétion non-specifiques, soit par des liens spécifiques mimant l'adhésion cellulaire. Par analyse d’image, nous différencions la réponses élastique, dans laquelle les gouttes se déforment à voisinage constant, de la réponse plastique, lors de laquelle les gouttes se réarrangent irréversiblement. Nous avons montré que la présence d'adhésion n'affectait pas la topologie globale des réarrangements, mais pouvait les ralentir localement. En conséquence, les gouttes présentent des déformations plus grandes et s'alignent avec la direction de l'élongation des tissus. Cela pourrait être la signature d'un processus de polarisation induit par l'adhésion dans les tissus en élongation. Le second aspect de mon projet, en collaboration avec Marie Breau, consiste à utiliser des émulsions biocompatibles comme senseurs de forces in vivo. En particulier, nous avons injecté des gouttes la placode olfactive d'embryons de poissons zèbres pour mesurer les forces associées à l'élongation des axones dans ce tissu. Les résultats obtenus suggèrent la présence de forces compressives qui pourraient contribuer à la croissance axonale. Des études plus approfondies seront menées afin de conclure sur l'origine et rôle de ces forces dans le développement de la placode olfactive. Les deux approches complémentaires utilisées pendant ma thèse permettent de lever le voile sur le rôle central des forces et de l'adhésion cellulaire dans la morphogénèse.

We have developed a method to produce aqueous microdroplets in an oil phase, based on the periodic extraction of a pending droplet across the oil/air interface. This interface forms a capillary trap inside which a droplet can be captured and detached. This process is found to be capillary- based and quasi-static. The droplet size and emission rate are independently governed by the injected volume per cycle and the extraction frequency. We find that the minimum droplet diameter is close to the injection glass capillary diameter and that variations in surface tension moderately perturb the droplet size. A theoretical model based on surface energy minimization in the oil/water/air phases was derived and captures the experimental results. This method enables robust, versatile and tunable production of microdroplets at low production rates.