The bioactivity of synthetic bone implants is highly impacted by their surface topography, especially by the presence of micro-patterns likely to generate cells growth guidance. In this study, laser machining technology was employed in order to produce controlled regular micro-patterns on dense calcium phosphate surfaces, without any contamination. The choice of the source was directed towards a femtosecond pulsed laser in order to limit the thermal impact of such a process and thus to avoid the unwanted phase transformations potentially induced by the temperature elevation. Beta tricalcium phosphate substrates with perfectly controlled micro-patterning and without any secondary phase were obtained by optimization of the process parameters (laser power, scanning speed, pulse frequency). The microstructural characteristics were investigated by microscopy (optical, confocal, scanning electron) and the phase identification was performed by X-ray diffraction. This work allowed highlighting the effects of the process parameters on the patterning. The high benefits of the laser treatment on wettability were shown by contact angle assays. Finally, the influence of surface micro-patterning on cell behavior was highlighted in vitro. This technique seems to provide an interesting alternative to conventional surface treatments of calcium phosphates.