Stimulation of bone repair with ultrasound: A review of the possible mechanic effects

In vivo and in vitro studies have demonstrated the positive role that ultrasound can play in the enhancement of fracture healing or in the reactivation of a failed healing process. We review the several options available for the use of ultrasound in this context, either to induce a direct physical effect (LIPUS, shock waves), to deliver bioactive molecules such as growth factors, or to transfect cells with osteogenic plasmids; with a main focus on LIPUS (or Low Intensity Pulsed Ultrasound) as it is the most widespread and studied technique. The biological response to LIPUS is complex as numerous cell types respond to this stimulus involving several pathways. Known to-date mechanotransduction pathways involved in cell responses include MAPK and other kinases signaling pathways, gap-junctional intercellular communication, up-regulation and clustering of integrins, involvement of the COX-2/PGE2, iNOS/NO pathways and activation of ATI mechanoreceptor. The mechanisms by which ultrasound can trigger these effects remain intriguing. Possible mechanisms include direct and indirect mechanical effects like acoustic radiation force, acoustic streaming, and propagation of surface waves, fluid-flow induced circulation and redistribution of nutrients, oxygen and signaling molecules. Effects caused by the transformation of acoustic wave energy into heat can usually be neglected, but heating of the transducer may have a potential impact on the stimulation in some in-vitro systems, depending on the coupling conditions. Cavitation cannot occur at the pressure levels delivered by LIPUS. In-vitro studies, although not appropriate to identify the overall biological effects, are of great interest to study specific mechanisms of action. The diversity of current experimental set-ups however renders this analysis very complex, as phenomena such as transducer heating, inhomogeneities of the sound intensity in the near field, resonances in the transmission and reflection through the culture dish walls and the formation of standing waves will greatly affect the local type and amplitude of the stimulus exerted on the cells. A future engineering challenge is therefore the design of dedicated experimental set-ups, in which the different mechanical phenomena induced by ultrasound can be controlled. This is a prerequisite to evaluate the biological effects of the different phenomena with respect to particular parameters, like intensity, frequency, or duty cycle. By relating the variations of these parameters to the induced physical effects and to the biological responses, it will become possible to derive an 'acoustic dose' and propose a quantification and cross-calibration of the different experimental systems. Improvements in bone healing management will probably also come from a combination of ultrasound with a 'biologic' components, e.g. growth factors, scaffolds, gene therapies, or drug delivery vehicles, the effects of which being potentiated by the ultrasound.