The effects of therapeutic ultrasound (US) on tissue healing processes in vivo are likely to involve US-induced changes in key cellular functions. However, these have not yet been clearly delineated and the present study has, therefore, examined the effects of a single 5-min CW exposure of 3.00-MHz US on the growth and functional activity of a human osteoblast-like cell line (MG63 cells) and human periodontal ligament (PDL) cells in vitro. Although cell proliferation was found to be largely unaffected by spatial average intensity (I(SA)) values of between 140-990 mW/cm(2), flow cytometry (FCM) analysis showed that there were pronounced and differential effects on cell function. Thus, bone-associated proteins were down-regulated, whereas collagen type I (COL I) was unaffected and fibronectin (FN) was up-regulated at low intensities in MG63 cells. In contrast, bone protein expression was found to be dose-dependent, and FN and COL I were down-regulated in PDL cells. These results show that US has potentially important effects on the functional activities of connective tissue cells in vitro, which could markedly influence tissue repair and regeneration processes in vivo.