Biomaterials design is important in the fields of regenerative medicine and tissue engineering, in particular to allow the long term expansion of stem cells and the engineering of scaffolds for tissue regeneration. Cell adhesion to biomaterials often plays a central role in regulating cell phenotype. It is emerging that physical properties of biomaterials, and more generally the microenvironment, regulate such behaviour. In particular, cells respond to nanoscale physical properties of their matrix. Understanding how such nanoscale physical properties control cell adhesion is therefore essential for biomaterials design. To this aim, a deeper understanding of molecular processes controlling cell adhesion, but also a greater control of matrix engineering is required. Such multidisciplinary approaches shed light on some of the fundamental mechanisms via which cell adhesions sense their nanoscale physical environment.

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 (

The culture of adherent cells is overwhelmingly relying on the use of solid substrates to support cell adhesion. Indeed, it is typically thought that relatively strong bulk mechanical properties (bulk moduli in the range of kPa to GPa) are essential to promote cell adhesion and, in turn, regulate cell expansion and fate decision. In this report, we show that adherent stem cells such as mesenchymal stem cells and primary keratinocytes can be cultured at the surface of liquid substrates and that this phenomenon is mediated by the assembly of polymer nanosheets at the liquid-liquid interface. We use interfacial rheology to quantify this assembly and demonstrate the strong mechanical properties of such nanosheets. Importantly, we show that cell adhesion to such quasi-2D materials is mediated by the classical integrin/acto-myosin machinery, despite the absence of bulk mechanical properties of the underlying liquid substrate. Finally, we show that stem cell proliferation and fate decision are also regulated by the mechanical properties of these self-assembled protein nanosheets. Liquid substrates offer attractive features for the culture of adherent cells and stem cells, and the development of novel stem cell technologies, such as liquid-liquid systems, are particularly well-adapted to automated parallel processing and scale up.

Thiol-ene radical coupling is increasingly used for the biofunctionalization of biomaterials. Thiol-ene chemistry presents interesting features that are particularly attractive for platforms requiring specific reactions with peptides or proteins and the patterning of cells, such as reactivity in physiological conditions and photoactivation. In this work, we synthesized alkene-functionalized (allyl and norbornene residues) antifouling polymer brushes (based on poly(oligoethylene glycol methacrylate)) and studied thiol-ene coupling with a series of thiols including cell adhesive peptides RGD and REDV. The adhesion of umbilical vein endothelial cells (HUVECs) to these interfaces was studied and highlighted the absence of specific integrin engagement to REDV, in contrast to the high level of cell spreading observed on RGD-functionalized polymer brushes. This revealed that αβ integrins (binding to REDV sequences) are not sufficient on their own to sustain HUVEC spreading, in contrast to αβ and αβ integrins. In addition, we photopatterned peptides at the surface of poly(oligoethylene glycol methacrylate) (POEGMA) brushes and characterized the quality of the resulting arrays by epifluorescence microscopy and atomic force microscopy (AFM). This allowed the formation of cell patterns and demonstrated the potential of thiol-ene based photopatterning for the design of cell microarrays.

Dynamic photo-responsive synthetic hydrogels offer important advantages for biomaterials design, from the ability to cure hydrogels and encapsulate cells in situ to the light-mediated control of cell spreading and tissue formation. We report the facile and effective photo-curing and photoremodelling of disulfide-crosslinked hyaluronic acid hydrogels, based on photo-oxidation of corresponding thiol residues and their radical-mediated photo-degradation. We find that the mechanical properties of disulfide hydrogels and the extent of their photo-remodelling can be tuned by controlling the photo-oxidation and photo-degradation reactions, respectively. This enables the photo-patterning of the mechanical properties of hydrogels, but also their self-healing and photomediated healing. Finally, we demonstrate the ability to encapsulate mesenchymal stromal cells within these materials and to regulate their protrusion and spreading in 3D matrices by controlling the mechanical properties of the disulfide networks. Therefore, synthetically accessible photoconfigurable disulfide hydrogels offer interesting opportunities for the design of soft biomaterials and the regulation of cell encapsulation and matrix remodelling for tissue engineering.