Non-invasive pulsed electromagnetic field (PEMF) stimulation is largely adopted in clinical practice for boosting the endogenous repair of bone fractures. However PEMF is applied empirically in clinics, without standardized guidelines, since many biological mechanisms and signalling pathways modulated by PEMF stimulation remain unknown. With a view to develop a reliable in vitro investigation methodology for studying the precise combination of physical parameters needed to reach a desired biological effect, the aim of this thesis was the optimization and validation of a versatile bioreactor that combines tunable direct perfusion and PEMF stimulation for in vitro investigating three-dimensional (3D) bone-like tissues under defined physical stimuli.