SummaryMechanical properties are cues for many biological processes in health or disease. In the heart, changes to the extracellular matrix composition and cross-linking result in stiffening of the cellular microenvironment during development. Moreover, myocardial infarction and cardiomyopathies lead to fibrosis and a stiffer environment, affecting cardiomyocyte behavior. Here, we identify that single cardiomyocyte adhesions sense simultaneous (fast oscillating) cardiac and (slow) non-muscle myosin contractions. Together, these lead to oscillating tension on the mechanosensitive adaptor protein talin on substrates with a stiffness of healthy adult heart tissue, compared with no tension on embryonic heart stiffness and continuous stretching on fibrotic stiffness. Moreover, we show that activation of PKC leads to the induction of cardiomyocyte hypertrophy in a stiffness-dependent way, through activation of non-muscle myosin. Finally, PKC and non-muscle myosin are upregulated at the costameres in heart disease, indicating aberrant mechanosensing as a contributing factor to long-term remodeling and heart failure.

Adherent cell culture typically requires cell spreading at the surface of solid substrates to sustain the formation of stable focal adhesions and assembly of a contractile cytoskeleton. However, a few reports have demonstrated that cell culture is possible on liquid substrates such as silicone and fluorinated oils, even displaying very low viscosities (0.77 cSt). Such behavior is surprising as low viscosity liquids are thought to relax much too fast (

Thiol-ene coupling enables the metal-free ultra-fast (seconds) crosslinking of polysiloxanes.

The mechanical properties of soft materials used in the biomedical field play an important role on their performance. In the field of tissue engineering, it is known that cells sense the mechanical properties of their environment, however some materials, such as Sylard 184 PDMS (poly(dimethylsiloxane)), have failed to elicit such response. It was proposed that differences in the mechanical properties of such soft materials, at different scales, could account for these discrepancies. Indeed, the variation in the elastic moduli obtained for soft materials characterised at different scales can span several orders of magnitude. This called for a side-by-side comparison of the mechanical behaviour of soft materials at different scales. Here we use indentation, rheology and atomic force microscopy nanoidentation (using different tip geometries) to characterise the mechanical properties of PDMS, poly(acrylamide) (PAAm) and carboxymethyl cellulose (CMC) hydrogels at different length scales. Our results highlight the importance of surface adhesion and the resulting changes in contact area, and sample microstructural heterogeneity, in particular for the mechanical characterisation of ultra-soft substrates at the nano- to micro-scale.

The mechanical properties of naturally-derived matrices and biomaterials are thought to play an important role in directing cell adhesion, spreading, motility, proliferation and differentiation. However, recent reports have indicated that cells may respond to local nanoscale physical cues, rather than bulk mechanical properties. We had previously reported that primary keratinocytes and mesenchymal stem cells did not seem to respond to the bulk mechanical properties of poly(dimethyl siloxane) (PDMS) substrates. In this study, we examine the mechanical properties of weakly crosslinked PDMS substrates and observe a liquid-like behaviour, with complete stress relaxation. We then report the observation that HaCaT cells, an epidermal cell line, proliferate readily at the surface of uncrosslinked liquid PDMS, as well as on low viscosity (0.77 cSt) fluorinated oil. These results are surprising, considering current views in the field of mechanotransduction on the importance of bulk mechanical properties, but we find that strong mechanical interfaces, presumably resulting from protein assembly, are formed at liquid-liquid interfaces for which cell adhesion and proliferation are observed. Hence our results suggest that cells sense the nanoscale mechanical properties of liquid-liquid interfaces and that such physical cues are sufficient to sustain the proliferation of adherent cells.