Material technology platforms able to modulate the communication with cells at the interface of biomaterials are being increasingly experimented. Progress in the fabrication of supports is simultaneously introducing new surface modification strategies aimed at turning these supports from passive to active components in engineered preparations. Among these platforms, polymer brushes are arising not only as coatings determining the physical and (bio)chemical surface properties of biomaterials, but also as smart linkers between surfaces and biological cues. Their peculiar properties, especially when brushes are synthesized by "grafting-from" methods, enable closer mimicking of the complex and heterogeneous biological microenvironments. Inspired by the growing interest in this field of materials science, we summarize here the most prominent and recent advances in the synthesis of "grafted-from" polymer brush surfaces to modulate the response of adhering cells.

If the binding strength of adhesive cues to the extracellular matrix (ECM) and the mechanisms involved in cell adhesion are synergistically correlated via a “mechanical” feedback, engineering of cue presentation at the ECM by designer macromolecules can enable control over cell–matrix interaction. Here, polymer brushes supporting fibronectin (FN) and presenting different grafted‐chain length to modulate cell interaction at ECM cell‐binding sites are exploited. Application of friction force microscopy allows us to estimate the lateral deformability and friction of oligoethylene glycol‐containing brushes. These parameters are demonstrated to regulate the adhesion of human mesenchymal stem cells (hMSCs), which adopt their morphology and form focal adhesions (FAs) responding to FN brush‐tether characteristics. Across a brush‐thickness gradient presenting uniform FN exposure, thin brushes stimulate cell spreading and the development of FAs. Conversely, thick and more laterally deformable polymer grafts induce a decrease in cell spreading and FA formation. A correlation between frictional forces experienced at the (macro)molecular scale and the behavior of stem cells has been found. This interaction can be clarified by exploring novel aspects of FFM, which demonstrates a powerful tool to dynamically probe the ECM environment and indirectly suggest a way to structure ECM in order to trigger specific cell responses.

The responsive properties of surface-grafted polymer films in aqueous media can be amplified by covalently layering thermosensitive brushes and hydrogels. This was demonstrated by synthesizing layers of linear poly(N-isopropylacrylamide) (PNIPAM) brushes, alternating with cross-linked, poly(hydroxyethyl)methacrylate (PHEMA) hydrogels via sequential surface-initiated atom-transfer radical polymerization (SI-ATRP) steps. Below the lower critical solution temperature (LCST) of PNIPAM, brush/hydrogel multilayered films swell similarly to linear PNIPAM homopolymer brushes, as measured by liquid ellipsometry. In contrast, above the LCST, the PHEMA hydrogel interlayer acts as stiffening element within the collapsed multilayered film, as monitored by atomic force microscopy (AFM) nanoindentation and lateral force microscopy (LFM). This translates into a 10-fold increase in Young's modulus by the collapsed, layered films compared to PNIPAM homopolymer analogues. The (macro)molecular continuity between the brush main chains and hydrogel constituents thus enables a chemically robust layering to form graded, quasi-3D grafted polymer architectures, which display a concerted and amplified temperature-triggered transition.

Coupling of rapid prototyping techniques and surface-confined polymerizations allows the fabrication of 3D multidirectional gradients of biomolecules within microporous scaffolds. The compositional gradients can be tailored by polymer-brush-assisted diffusion of protein solutions. This technique allows spatial control over stem cells manipulation within 3D environments.

Gradients of biomolecules on synthetic, solid substrates can efficiently mimic the natural, graded variation of properties of the extracellular matrix (ECM).

Poly(N-isopropylacrylamide) brushes with three different grafting densities were synthesized via surface-initiated atom-transfer radical polymerization on glass or on silicon substrates. The substrates were modified with monochlorosilane-based or trimethoxysilane-based atom-transfer radical polymerization initiators. Atomic force microscopy images showed detachment of brushes from the monochlorosilane-based system under cell culture conditions. In situ ellipsometry demonstrated the reversible swelling and collapse of the brushes as the temperature was varied across the lower critical solution temperature of poly(N-isopropylacrylamide) in pure water. The polymer brushes were evaluated as supporting substrates for MC-3T3 cell cultures. At 37°C (T>lower critical solution temperature), the seeded cells adhered, spread, and proliferated, whereas at 25°C (T<lower critical solution temperature), the cells detached from the surface. The low-density polymer brush showed the highest cell adhesion, featuring adhering cells with an elongated morphology.

Surface morphology and chemistry of polymers used as biomaterials, such as tissue engineering scaffolds, have a strong influence on the adhesion and behavior of human mesenchymal stem cells. Here we studied semicrystalline poly(ε-caprolactone) (PCL) substrate scaffolds, which exhibited a variation of surface morphologies and roughness originating from different spherulitic superstructures. Substrates were obtained by varying the parameters of the thermal processing, that is, crystallization conditions. The cells attached to these polymer substrates adopted different morphologies responding to variations in spherulite density and size. In order to decouple substrate topology effects on the cells, sub-100 nm bioadhesive polymer brush coatings of oligo(ethylene glycol) methacrylates were grafted from PCL and functionalized with fibronectin. On surfaces featuring different surface textures, dense and sub-100 nm thick brush coatings determined the response of cells, irrespective to the underlying topology. Thus, polymer brushes decouple substrate micro-/nanoscale surface topology and the adhesion of stem cells.

In this paper, we describe a method allowing one to perform three-dimensional displacement control in force spectroscopy by atomic force microscopy (AFM). Traditionally, AFM force curves are measured in the normal direction of the contacted surface. The method described can be employed to address not only the magnitude of the measured force but also its direction. We demonstrate the technique using a case study of angle-dependent desorption of a single poly(2-hydroxyethyl methacrylate) (PHEMA) chain from a planar silica surface in an aqueous solution. The chains were end-grafted from the AFM tip in high dilution, enabling single macromolecule pull experiments. Our experiments give evidence of angular dependence of the desorption force of single polymer chains and illustrate the added value of introducing force direction control in AFM.