RESUMO
Three patterned systems aiming at the control of mammalian cell behavior are presented. The determinant feature common to these systems is the spatial distribution of extracellular matrix (ECM) proteins (mainly collagen) on polymer substrates. This distribution differs from one system to another with respect to the scale at which it is affected, from the supracellular to the supramolecular scale, and with respect to the way it is produced. In the first system, the surface of polystyrene was oxidized selectively to form micrometer-scale patterns, using photolithography. Adsorption of ECM proteins in presence of a competitor was enhanced on the oxidized domains, allowing selective cell adhesion to be achieved. In the second system, electron beam lithography was used to engrave grooves (depth and width approximately 1 microm) on a poly(methyl methacrylate) (PMMA) substratum. No modification of the surface chemistry associated to the created topography could be detected. Cell orientation along the grooves was only observed when collagen was preadsorbed on the substratum. In the third system, collagen adsorbed on PMMA was dried in conditions ensuring the formation of a nanometer-scale pattern. Cell adhesion was enhanced on such patterned collagen layers compared to smooth collagen layers.
Assuntos
Adesão Celular/fisiologia , Técnicas de Cultura de Células/métodos , Materiais Revestidos Biocompatíveis/química , Proteínas da Matriz Extracelular/química , Fibroblastos/citologia , Fibroblastos/fisiologia , Polimetil Metacrilato/química , Adsorção , Animais , Adesão Celular/efeitos dos fármacos , Células Cultivadas , Materiais Revestidos Biocompatíveis/farmacologia , Colágeno/química , Proteínas da Matriz Extracelular/farmacologia , Humanos , Teste de Materiais , Camundongos , Ligação Proteica , Engenharia Tecidual/métodosRESUMO
High-resolution patterns of grooves have been made in poly(methyl methacrylate) films, PMMA, by an electron-beam microlithographic process. The surface of films processed over a large width was characterized in terms of chemical composition (X-ray photoelectron spectroscopy (XPS). time of flight secondary ion mass spectroscopy), wettability (sessile drop) and topography (atomic force microscopy). Collagen adsorption was also studied (radiocounting, XPS) as such or in competition with Pluronic F68. The chemical alteration of the surface induced by the electron-beam irradiation disappeared after the dissolution involved in the development process. W138 human fibroblasts cultivated on microgrooved substrata (grooves 1 microm deep and 0.5-10 microm wide) showed a strong orientation parallel to the grooves. The contact guidance is induced by the topography of the surface and not by the alternation of zones with different physico-chemical properties. It may be explained in terms of probability of successful substratum contact by cell protrusions.
