Biomedical subjects

Nikolaj Gadegaard

Publications and source records attributed to Nikolaj Gadegaard.

10 recordsLinked to original sources

PubMed2007

Nanotopographical stimulation of mechanotransduction and changes in interphase centromere positioning.

We apply a recently developed method for controlling the spreading of cultured cells using electron beam lithography (EBL) to create polymethylmethacrylate (PMMA) substrata with repeating nanostructures. There are indications that the reduced cell spreading on these substrata, compared with planar PMMA, results from a reduced adhesivity since there are fewer adhesive structures and fewer of their associated stress fibres. The reduced cell spreading also results in a reduced nuclear area and a closer spacing of centrosomes within the nucleus, suggesting that the tension applied to the nucleus is reduced as would be expected from the reduction in stress fibres. In order to obtain further evidence for this, we have used specific inhibitors of components of the cytoskeleton and have found effects comparable with those induced by the new substrata. We have also obtained evidence that these subtrata result in downregulation of gene expression which suggests that this may be due to the changed tension on the nucleus: an intriguing possibility that merits further investigation.

Cell Adhesion↗

PubMed2006

Group analysis of regulation of fibroblast genome on low-adhesion nanostructures.

Nanotopographical material modification represents a possible way of producing surfaces that influence cellular adhesion for biomaterials purposes. Here, two low-adhesion surfaces are studied with human genome microarrays (120nm diameter pits produced by electron beam lithography and 11nm high columns produced by colloidal lithography). In order to present the large numbers of results produced in a succinct and easy to understand manner, two types of recent bioinformatics methods were used; iterative group analysis and Ingenuity pathway analysis. These methods allowed the easy comparison of the nanomaterials and showed large-scale changes in areas of extracellular matrix, cell signalling and inflammation. The results demonstrate that whilst modes of cellular response to low-adhesion materials are similar, the more adhesion is reduced, the further 'shut-down' of critical cellular activities is observed. We also feel that the analysis used could be of interest to biomaterials scientists looking for easy ways to display microarray data efficiently.

Adhesiveness↗

PubMed2006

Air-trapping on biocompatible nanopatterns.

The occurrence of air-trapping inside poly-eta-caprolactone nanopits was investigated by measuring the contact angles of water droplets on a set of defined nanotopographies. It is shown that the advancing angles follow the Cassie-Baxter theory, thus revealing the presence of air bubbles inside the biodegradable nanopatterns. The importance of these observations for the definition of hydrophilicity/hydrophobicity and in the context of in vitro cell behavior is discussed.

Absorption↗

PubMed2006

Nanoprinting onto cells.

Growing cells on surfaces bearing nanotopography signals makes many changes in cell gene expression and downstream changes in phenotype but the mechanisms for this have, so far, been obscure. We consider the question of whether the topography directly nanoimprints onto the cell as a component of the signal transduction system. Evidence we present from SEM, TEM and fluorescence detection of the arrangements of cytoskeletal components is consistent with the possibility that cells are nanoimprinted by the substrate. The nanoprinting does not interfere with integrin-mediated adhesion processes and may perhaps work through them. Time-lapse video studies of cells moving from areas bearing nanotopography to flat areas and vice versa suggests that the nanoimprinting takes 1-6h to appear on the cell and a similar time to disappear when the cell moves from a flat surface to a nanotopographic one and back. This nanoprinting of cells would appear to be a novel type of cell signalling.

Cell Movement↗

PubMed2006

Fluorescent aromatic platforms for cell patterning.

This paper describes a simple experimental method of patterning fluorescent organic dyes, fluorescein, and rhodamine on gold substrates by microcontact printing techniques. The development of this step-by-step protocol has allowed us to prepare striped and squared micropatterns with poly(ethylene glycol) (PEG) linkers terminated by these fluorophores using a fast, easy, and inexpensive technique. Although the rest of the surface was covered with aliphatic molecules (methyl terminated), human fibroblasts demonstrated an unexpected response, aligning themselves according to the aromatic patterns, despite the presence of PEG, which is a cell resistant molecule, in the fluorescent regions.

Cell Adhesion↗

PubMed2005

Evaluating the influence of deposition conditions on solvation of reactive conducting polymers with neutron reflectivity.

We describe in situ neutron reflectivity (NR) and RAIRS studies of the chemical modification of films of a polypyrrole-based conducting polymer derived from the pentafluorophenyl ester of poly(pyrrole-N-propanoic acid) (PFP) electrodeposited on electrode surfaces. We explore the role of the solvent in controlling the rate of reaction with solution-based nucleophiles (amines, which react with the ester to form amides). By varying the identity of the solvent (water vs acetonitrile) and the neutron contrast (deuteration), we find that both the identity of the solvent and its population within the film are paramount in determining chemical reactivity and electroactivity. IR signatures allow monitoring of the reaction of solution-based amine-tagged species such as amino-terminated poly(propylene glycol), ferrocene ethylamine, and lysine with film-based ester functionalities: the carbonyl bands show ester/amide interconversion and some hydrolysis to acid. Time-dependent spectral analysis shows marked variations in reaction rate with (i) (co-)polymer composition (replacement of some fluorinated ester-functionalized pyrrole with unfunctionalized pyrrole), (ii) the solvent to which the polymer film is exposed, and (iii) the rate of polymer deposition. NR data provide solvent profiles as a function of distance perpendicular to the interface, the variations of which provide an explanation for film reactivity patterns. Homopolymer films are relatively hydrophobic, thus hindering reaction with species present in water solutions. Incorporating pyrrole groups raises the solvent population-dramatically for water-thereby facilitating entry and reaction of aqueous-based lysine. Changing film deposition rate yields films with different absolute levels of solvent and reactivity patterns that are dependent on the size of the reactant molecules: more rapid deposition of polymer gives films with a more open structure leading to a higher solvent content and thence increased reactivity. These results, supported by XPS and AFM data, allow assembly of composition-structure-reactivity correlations, in which the controlling feature is film solvation.

Electrochemistry↗

PubMed2005

Superhydrophobicity and superhydrophilicity of regular nanopatterns.

The hydrophilicity, hydrophobicity, and sliding behavior of water droplets on nanoasperities of controlled dimensions were investigated experimentally. We show that the "hemi-wicking" theory for hydrophilic SiO(2) samples successfully predicts the experimental advancing angles and that the same patterns, after silanization, become superhydrophobic in agreement with the Cassie-Baxter and Wenzel theories. Our model topographies have the same dimensional scale of some naturally occurring structures that exhibit similar wetting properties. Our results confirm that a forest of hydrophilic/hydrophobic slender pillars is the most effective superwettable/water-repellent configuration. It is shown that the shape and curvature of the edges of the asperities play an important role in determining the advancing angles.

Journal Article↗

PubMed2004

Dynamic in situ electrochemical neutron reflectivity measurements.

We report the first dynamic in situ electrochemical neutron reflectivity (NR) measurements on electroactive films. By using a boxcar integration strategy within a cyclic voltammetric experiment, it is possible to acquire neutron reflectivity data associated with narrow, defined windows of potential. Accumulation of data from repetitive cycles allows one to build up potential- (time-) resolved profiles. The effective time resolution is now on the order of seconds, as compared to ca. 1 h using conventional methodology, making in situ NR a practical technique for dynamic electrochemical studies. Illustrative data for polyvinylferrocene films reveal hysteresis in (de)swelling, incomplete desolvation upon reduction, and transient salt retention, all of which respond to time-scale variations.

Journal Article↗

PubMed2004

Investigating filopodia sensing using arrays of defined nano-pits down to 35 nm diameter in size.

In order for cells to react to topography, they must be able to sense shape. When considering nano-topography, these shapes are much smaller than the cell, but still strong responses to nano-topography have been seen. Filopodia, or microspikes, presented by cells at their leading edges are thought to be involved in gathering of special information. In order to investigate this, and to develop an understanding of what size of feature can be sensed by cells, morphological observation (electron and fluorescent microscopy) of fibroblasts reacting to nano-pits with 35, 75 and 120 nm diameters has been used in this study. The nano-pits are especially interesting because unlike many of the nanofeatures cited in the literature, they have no height for the cells to react to. The results showed that cell filopodia, and retraction fibres, interacted with all pit sizes, although direct interaction was hard to image on the 35 nm pits. This suggests that cells are extremely sensitive to their nanoevironment and that should be taken in to consideration when designing next-generation tissue engineering materials. We suggest that this may occur through nanocontact guidance as filopodia are moved over the pits.

Cells, Cultured↗

PubMed2004

The effects of collagen type I topography on myoblasts in vitro.

Cells respond to a variety of cues from their environment, which can include chemical, mechanical, and topographical signals. The differentiation of myoblasts requires a combination of signals. Myoblast fusion is strongly influenced by the chemical nature of the surrounding matrix and can be affected by mechanical stimulation. Studies also have shown that a large variety of cell types also are influenced by details of surface topography of a substrate as small as 44 nm. Cells grown on a collagen-coated surface differentiate more readily than those grown in the absence of the extracellular matrix protein. It is not known whether the effects of myoblast interaction with collagen are due solely to chemical interactions or if myoblasts also respond to the topography of collagen type I fibers. To determine the importance of collagen-generated topographical signals on myoblast development, cells were cultured and differentiated in vitro on surfaces that had been coated with either soluble collagen type I or fibrous collagen type I. Both surfaces present the same chemical interactions, but the additional topographical signals lead to differences in cell morphology, adhesion, spreading rates and, proliferation. Cells on the fibrous form of collagen are more stellate, form more adhesion plaques, spread faster, and proliferate at a faster, rate than cells on a surface of soluble collagen. Our data indicate that topographical signals play a role in early muscle development, but that other or additional signaling pathways regulate differentiation.

Animals↗