Morphology, proliferation, alignment, and new collagen synthesis of mesenchymal stem cells on a microgrooved collagen membrane.

The topographic cues of the extracellular matrix may have significant effects upon cellular behavior, such as adhesion, spreading, migration, proliferation, differentiation, and in particular, morphology and orientation. In this study, we examined the effects of microgrooved collagen membrane (MCM) on mesenchymal stem cell (MSC) behavior. The MCM (9 μm in periodicity, and 1-2 μm in depth) was fabricated on an untreated (nonpolar) and smooth polystyrene substrate, based on the absorption and self-assembly properties of collagen on the polystyrene substrate. Methyl thiazolyl tetrazolium assay revealed that cell proliferation on the MCM was enhanced compared with the smooth collagen membrane at day 2. Qualitative observation of MSC behavior using confocal laser scanning microscopy and scanning electron microscopy showed that MSCs grew with a highly elongated morphology and were aligned strictly along the direction of the microgrooves. Additionally, scanning electron microscopy revealed the oriented cells produced a collagenous matrix on the MCM that had a preferential orientation, whereas the collagenous matrix produced by randomly oriented MSCs on the smooth collagen membrane was disorganized. Future studies should investigate the fabrication of oriented topographical substrates, based on the natural biomaterial collagen, to guide cell alignment and oriented growth along definite directions. These substrates may help produce aligned collagenous matrices that could have good potential for the production of tissue substitutes.