16 research outputs found
Spatially controlled cell adhesion on three-dimensional substrates
The microenvironment of cells in vivo is defined by spatiotemporal patterns of chemical and biophysical cues. Therefore, one important goal of tissue engineering is the generation of scaffolds with defined biofunctionalization in order to control processes like cell adhesion and differentiation. Mimicking extrinsic factors like integrin ligands presented by the extracellular matrix is one of the key elements to study cellular adhesion on biocompatible scaffolds. By using special thermoformable polymer films with anchored biomolecules micro structured scaffolds, e.g. curved and µ-patterned substrates, can be fabricated. Here, we present a novel strategy for the fabrication of µ-patterned scaffolds based on the “Substrate Modification and Replication by Thermoforming” (SMART) technology: The surface of a poly lactic acid membrane, having a low forming temperature of 60°C and being initially very cell attractive, was coated with a photopatterned layer of poly(L-lysine) (PLL) and hyaluronic acid (VAHyal) to gain spatial control over cell adhesion. Subsequently, this modified polymer membrane was thermoformed to create an array of spherical microcavities with diameters of 300 µm for 3D cell culture. Human hepatoma cells (HepG2) and mouse fibroblasts (L929) were used to demonstrate guided cell adhesion. HepG2 cells adhered and aggregated exclusively within these cavities without attaching to the passivated surfaces between the cavities. Also L929 cells adhering very strongly on the pristine substrate polymer were effectively patterned by the cell repellent properties of the hyaluronic acid based hydrogel. This is the first time cell adhesion was controlled by patterned functionalization of a polymeric substrate with UV curable PLL-VAHyal in thermoformed 3D microstructures
Processing and characterization of chitosan microspheres to be used as templates for layer-by-layer assembly
Chitosan (Ch) microspheres have been developed
by precipitation method, cross-linked with glutaraldehyde
and used as a template for layer-by-layer (LBL)
deposition of two natural polyelectrolytes. Using a LBL
methodology, Ch microspheres were alternately coated with
hyaluronic acid (HA) and Ch under mild conditions. The
roughness of the Ch-based crosslinked microspheres was
characterized by atomic force microscopy (AFM). Morphological
characterization was performed by environmental
scanning electron microscopy (ESEM), scanning
electron microscopy (SEM) and stereolight microscopy.
The swelling behaviour of the microspheres demonstrated
that the ones with more bilayers presented the highest water
uptake and the uncoated cross-linked Ch microspheres
showed the lowest uptake capability. Microspheres presented
spherical shape with sizes ranging from 510 to
840 lm. ESEM demonstrated that a rougher surface with
voids is formed in multilayered microspheres caused by the
irregular stacking of the layers. A short term mechanical
stability assay was also performed, showing that the LBL
procedure with more than five bilayers of HA/Ch over Ch
cross-linked microspheres provide higher mechanical
stability
Framework for understanding marine ecosystem health
Although the terms ‘health’ and ‘healthy’ are often applied to marine ecosystems and communicate information about holistic condition (e.g. as required by the Ecosystem Approach), their meaning is unclear. Ecosystems have been understood in various ways, from non-interacting populations of species to complex integrated systems. Health has been seen as a metaphor, an indicator that aggregates over system components, or a non-localized emergent system property. After a review, we define good ecosystem health as: ‘the condition of a system that is self-maintaining, vigorous, resilient to externally imposed pressures, and able to sustain services to humans. It contains healthy organisms and populations, and adequate functional diversity and functional response diversity. All expected trophic levels are present and well interconnected, and there is good spatial connectivity amongst subsystems.’ We equate this condition with good ecological or environmental status, e.g. as referred to by recent EU Directives. Resilience is central to health, but difficult to measure directly. Ecosystems under anthropogenic pressure are at risk of losing resilience, and thus of suffering regime shifts and loss of services. For monitoring whole ecosystems, we propose an approach based on ‘trajectories in ecosystem state space’, illustrated with time-series from the northwestern North Sea. Change is visualized as Euclidian distance from an arbitrary reference state. Variability about a trend in distance is used as a proxy for inverse resilience. We identify the need for institutional support for long time-series to underpin this approach, and for research to establish state space co-ordinates for systems in good health