Protease-sensitive hydrogels that recapitulate the mechanisms of cell-driven enzymatic remodelling of the natural extracellular matrix (ECM) have been gaining popularity as artificial 3D cell-microenvironments.Here, the matrix metalloproteinase (MMP)-sensitive peptide Pro-Val-Gly-Leu-Iso-Gly (PVGLIG) was double-end grafted to alginate forming water-soluble PVGLIG-alginate conjugates. The PVGLIG peptide was synthesized as a Fluorescence Resonance Energy Transfer (FRET) sensor and showed to be a good substrate for MMP-2, MMP-9, MMP-13 and MMP-14. After demonstrating that human MSC (hMSC) expressed both MMP-2 and MMP-14 under basal and osteogenic in vitro conditions, the effect of 3Dculture within MMP-sensitive alginate hydrogels on hMSC behaviour was addressed. In situ-forming alginate hydrogels containing only cell-adhesive RGD peptides (RGD-alginate, MMP-insensitive) or both peptides (PVGLIG/RGD-alginate, MMP-sensitive) were used. Cell-matrix and cell-cell interactions were enhanced in hMSC-laden MMP-sensitive alginate hydrogels, as compared to MMP-insensitive hydrogels with identical viscoelastic and microstructural properties. hMSC underwent osteogenic differentiation in both types of matrices. However, the presence of PVGLIG stimulated the secretion of proteases (most likely MMP-2) by hMSC, in both undifferentiated and differentiated cultures. By using the FRET sensor, it was possible to demonstrate that the cocktail of hMSC-secreted MMPs was effectively active in cleaving the PVGLIG motif. Protease-sensitive alginates can be used to create cell-responsive 3D microenvironments and offer promise as injectable carriers for therapeutic hMSC-delivery.

No abstract

No abstract

We simulate the isothermal, incompressible, time-dependent flow of Boger fluids, using the FENE-CR constitutive model, in the optimized shape cross-slot extensional rheometer [Haward et al., Phys. Rev. Lett. 109, 128301 (2012)]. We uncover a family of predominantly elastic instabilities resulting from the interaction of stationary asymmetric and time-dependent flow transitions. The superposition of these instabilities, with varying degrees of relative amplitude, produces five distinct flow regimes which are classified by their elasticity, suggesting that the dynamic system is situated in the vicinity of a triple point in the inertia-elasticity-viscosity parameter space. A detailed characterization of the various associated modes of instability is provided. Spectral analysis of the first component of the velocity vector suggests that the flow regimes featuring loss of birefringence strand integrity become chaotic and locally turbulent near the center of the cross-slot, above a certain flow rate threshold.