Our previous studies have shown that on human periosteal cells, low-intensity pulsed ultrasound (LIPUS) has an immediate stimulatory effect whereas extracorporeal shockwaves (ESW) have an delayed stimulatory effect. Therefore, we hypothesized that a combined ESW and LIPUS treatment might provide additive or synergistic effects on periosteal cells, by using ESW to trigger a biological activity while using LIPUS to maintain the stimulated activity. Human periosteal cells were subjected to a single session of ESW treatment on day 0 and/or daily LIPUS treatments or no treatment (control). The cell viability, proliferation, and alkaline phosphatase activity on day 6 and day 18 as well as matrix mineralization on day 35 were measured. Results revealed that LIPUS alone had early positive effects on the activities on day 6 only. In contrast, ESW alone had an early destructive effect but exerted delayed stimulatory effects on the cellular activities on day 18. The combined treatment of ESW plus LIPUS produced effects that were comparable to the ESW treatment alone. Although these findings suggest that ESW and LIPUS stimulate the periosteal cells in two different ways and at different times, their additive or synergistic effects could not be proven.