Olivier Ronsin

Transcription factor EGR1 directs tendon differentiation and promotes tendon repair

Tendon formation and repair rely on specific combinations of transcription factors, growth factors, and mechanical parameters that regulate the production and spatial organization of type I collagen. Here, we investigated the function of the zinc finger transcription factor EGR1 in tendon formation, healing, and repair using rodent animal models and mesenchymal stem cells (MSCs). Adult tendons of Egr1-/- mice displayed a deficiency in the expression of tendon genes, including Scx, Col1a1, and Col1a2, and were mechanically weaker compared with their WT littermates. EGR1 was recruited to the Col1a1 and Col2a1 promoters in postnatal mouse tendons in vivo. Egr1 was required for the normal gene response following tendon injury in a mouse model of Achilles tendon healing. Forced Egr1 expression programmed MSCs toward the tendon lineage and promoted the formation of in vitro-engineered tendons from MSCs. The application of EGR1-producing MSCs increased the formation of tendon-like tissues in a rat model of Achilles tendon injury. We provide evidence that the ability of EGR1 to promote tendon differentiation is partially mediated by TGF-β2. This study demonstrates EGR1 involvement in adult tendon formation, healing, and repair and identifies Egr1 as a putative target in tendon repair strategies.

Self-healing slip pulses and the friction of gelatin gels

Abstract:We present an extensive experimental study and scaling analysis of friction of gelatin gels on glass. At low driving velocities, sliding occurs via propagation of periodic self-healing slip pulses whose velocity is limited by collective diffusion of the gel network. Healing can be attributed to a frictional instability occurring at the slip velocity V = Vc. For V > Vc, sliding is homogeneous and friction is ruled by the shear-thinning rheology of an interfacial layer of thickness of order the (nanometric) mesh size, containing a solution of polymer chain ends hanging from the network. In spite of its high degree of confinement, the rheology of this system does not differ qualitatively from known bulk ones. The observed ageing of the static friction threshold reveals the slow increase of adhesive bonding between chain ends and glass. Such structural ageing is compatible with the existence of a velocity-weakening regime at velocities smaller than Vc, hence with the existence of the healing instability.

Magic angles and cross-hatching instability in hydrogel fracture.

The full 2D analysis of roughness profiles of fracture surfaces resulting from quasistatic crack propagation in gelatin gels reveals an original behavior characterized by (i) strong anisotropy with maximum roughness at V-independent symmetry-preserving angles and (ii) a subcritical instability leading, below a critical velocity, to a cross-hatched regime due to straight macrosteps drifting at the same magic angles and nucleated on crack-pinning network inhomogeneities. Step height values are determined by the width of the strain-hardened zone, governed by the elastic crack blunting characteristic of soft solids with breaking stresses much larger than low strain moduli.

A convective instability mechanism for quasistatic crack branching in a hydrogel

Experiments on quasistatic crack propagation in gelatin hydrogels reveal a new branching instability triggered by wetting the tip opening with a drop of aqueous solvent less viscous than the bulk one. We show that the emergence of unstable branches results from a balance between the rate of secondary crack growth and the rate of advection away from a non-linear elastic region of size G/E , where G is the fracture energy and E the small strain Young modulus. We build a minimal, predictive model that combines mechanical characteristics of this mesoscopic region and physical features of the process zone. It accounts for the details of the stability diagram and lends support to the idea that non-linear elasticity plays a critical role in crack front instabilities.

Interplay between Shear Loading and Structural Aging in a Physical Gelatin Gel

We show that the aging of the mechanical relaxation of a gelatin gel exhibits the same scaling phenomenology as polymer and colloidal glasses. In addition, gelatin is known to exhibit logarithmic structural aging (stiffening). We find that stress accelerates this process. However, this effect is definitely irreducible to a mere age shift with respect to natural aging. We suggest that it is interpretable in terms of elastically aided elementary (coil --> helix) local events whose dynamics gradually slows down as aging increases geometric frustration.

Chemo-osmotically driven inhomogeneity growth during the enzymatic gelation of gelatin

We present an extensive study of the enzyme-mediated, isothermal formation of covalently cross-linked gelatin gels. We find that the enzymatic activity in the forming network is drastically reduced compared with that in solution, and show that this can be attributed to the growing level of cross-link induced geometric constraints which impede translational and rotational motions. Thanks to the slowness of these kinetics, we monitor the concomitant build-up of the shear modulus G′ and of the optical turbidity , which indicates that gelation is associated with the development of a high level of inhomogeneity. We find that, as the gelatin concentration cG is varied, the levels of G′ and are strongly anti-correlated. Moreover, the lower cG, the more precocious the emergence of . We are able to analyze inhomogeneity development in terms of the amplification of structural fluctuations via the coupling between the kinetics of the cross-linking reaction and the osmotic flow driven by swelling pressure fluctuations. We expect this positive feedback mechanism to be efficient in any slow, irreversible gelation process.

Two-step build-up of a thermoreversible polymer network: From early local to late collective dynamics

We probe the mechanisms at work in the build-up of thermoreversible gel networks, with the help of hybrid gelatin gels containing a controlled density of irreversible, covalent crosslinks (CLs), which we quench below the physical gelation temperature. The detailed analysis of the dependence on covalent crosslink density of both the shear modulus and optical activity evolutions with time after quench enables us to identify two stages of the physical gelation process, separated by a temperature-dependent crossover modulus: (i) an early nucleation regime during which rearrangements of the triple-helix CLs play a negligible role, and (ii) a late, logarithmic aging one, which is preserved, though slowed down, in the presence of irreversible CLs. We show that aging is fully controlled by rearrangements and discuss the implication of our results in terms of the switch from an early, local dynamics to a late, cooperative long-range one.

Preferential hydration fully controls the renaturation dynamics of collagen in water-glycerol solvents

Abstract.Glycerol is one of the additives which stabilize collagen, as well as globular proteins, against thermally induced denaturation --an effect explained by preferential hydration, i.e. by the formation, in water/glycerol solvents, of a hydration layer whose entropic cost favors the more compact triple-helix native structure against the denatured one, gelatin. Quenching gelatin solutions promotes renaturation which, however, remains incomplete, as the formation of a gel network gives rise to growing topological constraints. So, gelatin gels exhibit glass-like dynamical features such as slow aging of their shear modulus and stretched exponential stress relaxation, the study of which gives us access to the re(de)naturation dynamics of collagen. We show that this dynamics is independent of the bulk solvent viscosity and controlled by a single parameter, the undercooling

Nucleation and Propagation of Quasi-Static Interfacial Slip Pulses

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The physics and chemistry of silica-in-silicates nanocomposite hydrogels and their phycocompatibility

ΔT below the glycerol-concentration-dependent denaturation temperature. This provides direct proof of i) the presence of a nanometer thick, glycerol-free hydration layer, ii) the high locality of the kinetically limiting process governing renaturation.Graphical abstract