Single-cell micropatterning of osteocytes and nanobiological characterization

In this work, we developed star-shaped micropatterns for single-cell growth and analysis of dendritic osteocytes. Micropatterning improves the visualisation and quantitative analysis of adhered cells, which precisely adapt their cytoskeleton architecture to the geometry of their environment. This allows cell normalisation and detection of subtle changes in cell response to perturbations. Two star-shaped micropatterns containing dendrite-like adhesive lines were fabricated using a combination of scanning probe lithography and microcontact printing. Osteocyte adhesion and growth was optimised. The cytoskeletal architecture was analysed using fluorescence microscopy and AFM stiffness tomography. Single osteocyte cells were successfully adhered and spread up to the edges of the nanosized adhesive lines of the micropatterns. Stress fibres were observed by staining with fluorescently-labelled phalloidin. AFM force volume spectroscopy was used to determine the cytoskeleton stiffness of the cells on the micropatterns.