Patch-clamp recording techniques were used to examine the direct effects of mechanical stimulation on ion channel activity in human osteoblast-like osteosarcoma cells. Three classes of mechanosensitive ion channels were present and could be distinguished on the basis of conductance, ionic selectivity, and sensitivity to membrane tension. The largest conductance channel (160 pS) was K(+)-selective and showed both a decrease in long closed interval duration and an increase in burst length with increasing membrane tension. For low applied pressures, there was an e-fold increase in the probability of this channel being open (Popen) for every 3.4 cm2 Hg change in pressure. Two additional pressure-dependent channels had smaller conductances, i.e., 60 pS and 20 pS; the 60 pS channel appeared to be non-selective for cations. We propose that one or more of these mechanosensitive channels is involved in the response of bone to mechanical loading.