Mesenchymal stem cells (MSCs) differentiate to cells that are part of tissues, such as bone and cartilage, both during ontogeny and for the repair/regeneration and renewal of tissues during postnatal life. Accordingly, MSCs can be a powerful tool for regenerative medicine and tissue engineering interventions in musculoskeletal deficiencies. MSCs found in the bone marrow and connective tissues are exposed to biochemical and biophysical factors that modulate their biological behavior. Given that biochemical factors have been widely studied, research on the effect of biophysical stimuli on the differentiation of MSCs is scarce. An overview of the impact of biophysical factors on MSCs osteogenic differentiation could shed light on some of the signals to consider when developing new cell therapies or tissue engineering products for the treatment of bone traumatic defects and diseases. The aim of this review is to provide an overview of the effect of biophysical stimuli on osteogenic differentiation. It focuses on reviewing what has been published on the effect of compression, tension, hydrostatic pressure, shear stress, electromagnetic fields, low-intensity ultrasound, and niche signaling – including topography and substrate stiffness, paracrine and juxtracrine regulation – on MSCs osteogenic differentiation. The knowledge gained from studying how MSCs respond to mechanical forces has the potential to drive the creation of more advanced treatments for conditions where bones are injured. These therapies could rely on either physical methods or a combination of physical and chemical approaches, offering new possibilities for improving bone health.