Mucus from human bronchial epithelial cultures: rheology and adhesion across length scales.

Mucus is a viscoelastic aqueous fluid that participates in the protective barrier of many mammals' epithelia. In the airways, together with cilia beating, mucus rheological properties are crucial for lung mucociliary function, and, when impaired, potentially participate in the onset and progression of chronic obstructive pulmonary disease (COPD). Samples of human mucus collected in vivo are inherently contaminated and are thus poorly characterized. Human bronchial epithelium (HBE) cultures, differentiated from primary cells at an air-liquid interface, are highly reliable models to assess non-contaminated mucus. In this paper, the viscoelastic properties of HBE mucus derived from healthy subjects, patients with COPD and from smokers are measured. Hallmarks of shear-thinning and elasticity are obtained at the macroscale, whereas at the microscale mucus appears as a heterogeneous medium showing an almost Newtonian behaviour in some extended regions and an elastic behaviour close to boundaries. In addition, we developed an original method to probe mucus adhesion at the microscopic scale using optical tweezers. The measured adhesion forces and the comparison with mucus-simulants rheology as well as mucus imaging collectively support a structure composed of a network of elastic adhesive filaments with a large mesh size, embedded in a very soft gel.

Design of vesicle prototissues as a model for cellular tissues.

Synthesizing biomimetic prototissues with predictable physical properties is a promising tool for the study of cellular tissues, as they would enable to test systematically the role of individual physical mechanisms on complex biological processes. The aim of this study is to design a biomimetic cohesive tissue with tunable mechanical properties by the controlled assembly of giant unillamelar vesicles (GUV). GUV-GUV specific adhesion is mediated by the inclusion of the streptavidin-biotin pair, or DNA complementary strands. Using a simple assembly protocol, we are capable of synthesizing vesicle prototissues of spheroidal or sheet-like morphologies, with predictable cell-cell adhesion strengths, typical sizes, and degree of compaction.

Culture of human neurospheres in 3D scaffolds for developmental neurotoxicity testing.

Human neural progenitor cells cultured as neurospheres are a promising tool for developmental neurotoxicity testing in vitro. In order to obtain a human cell-based tissue culture system as close to the organ as possible, it is desirable to improve the spatial organization of the "Neurosphere Assay" and use 3D scaffolds to better mimic the in vivo three dimensional cell microenvironment. For this reason we have established the conditions for short-term culture (up to 6 days) in matrigel or in IKVAV-3 peptide-functionalized hydrogels, and for long-term culture (>25 days) in IKVAV-3 peptide-functionalized hydrogels showing that these conditions support human neural progenitor cells' migration, differentiation to neurons and formation of neuronal networks. Moreover, we assessed if neurospheres grown in 3D scaffolds allow for developmental neurotoxicity compound testing. At concentrations not affecting cell viability the known developmental neurotoxic compound MeHgCl inhibits migration of human neural progenitor cells grown in 3D scaffolds with a higher potency than when the same cells are cultured on a laminin-coated surface as secondary 3D structures. Thus, this work opens the door to functional assessment of compound effects on short- and long-term cultured human neurospheres embedded in 3D scaffolds for developmental neurotoxicity testing.

The stabilizing effect of shear thinning on the onset of purely elastic instabilities in serpentine microflows.

We determine both experimentally and numerically the onset of elastic flow instabilities in viscoelastic polymer solutions with different levels of shear thinning. Previous experiments realized in microfluidic serpentine channels using dilute polymeric solutions showed that the onset of elastic instabilities strongly depends on the channel curvature. The scaling dependence is well captured by the general instability scaling criterion proposed by Pakdel and McKinley [Phys. Rev. Lett., 1996, 76, 2459:1-4]. We determine here the influence of fluid shear thinning on the onset of such purely-elastic flow instabilities. By testing a set of polyethylene oxide solutions of high molecular weight at different polymer concentrations in microfluidic serpentine channels we observe that shear thinning has a stabilizing effect on the microfluidic flow. Three-dimensional numerical simulations performed using the White-Metzner model predict similar trends, which are not captured by a simple scaling analysis using the Pakdel-McKinley criterion.

Asymmetric stochastic switching driven by intrinsic molecular noise.

Low-copy-number molecules are involved in many functions in cells. The intrinsic fluctuations of these numbers can enable stochastic switching between multiple steady states, inducing phenotypic variability. Herein we present a theoretical and computational study based on Master Equations and Fokker-Planck and Langevin descriptions of stochastic switching for a genetic circuit of autoactivation. We show that in this circuit the intrinsic fluctuations arising from low-copy numbers, which are inherently state-dependent, drive asymmetric switching. These theoretical results are consistent with experimental data that have been reported for the bistable system of the gallactose signaling network in yeast. Our study unravels that intrinsic fluctuations, while not required to describe bistability, are fundamental to understand stochastic switching and the dynamical relative stability of multiple states.