Michel Klein Gunnewiek

Thin Polymer Brush Decouples Biomaterial’s Micro-/Nanotopology and Stem Cell Adhesion

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.

Stability and Cell Adhesion Properties of Poly(N-isopropylacrylamide) Brushes with Variable Grafting Densities

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.

Creeping Proteins in Microporous Structures: Polymer Brush-Assisted Fabrication of 3D Gradients for Tissue Engineering

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.

Mimicking natural cell environments: design, fabrication and application of bio-chemical gradients on polymeric biomaterial substrates

Gradients of biomolecules on synthetic, solid substrates can efficiently mimic the natural, graded variation of properties of the extracellular matrix (ECM). Such gradients represent accessible study platforms for the understanding of cellular activities, and they also provide functional supports for tissue engineering (TE). This review describes the most relevant methods to produce 2-dimensional (2D) as well as 3-dimensional (3D) gradient supports for cell manipulations, and also addresses the response of cells from different origins when seeded on these constructs. The fabrication strategies summarized encompass the combination of polymer and surface chemistries, micro- and nano-engineering construction strategies and biotechnological approaches. This multidisciplinary scheme has enabled the design and realization of diverse, synthetic supports as cellular environments, spanning from the first gradient self-assembled monolayer (SAM) to multilayers, and hybrid constructs mimicking the complexity of natural tissue environments. The standing challenge is bringing these advances in the fabrication of supports to a dynamic functioning in space and time, towards the successful imitation of the most complex bio-chemical system ever studied: our body.