We have reported that low-intensity pulsed ultrasound (LIPUS) accelerated periodontal wound healing in experimental alveolar bone defects in dogs, however, the exact mechanism is unknown. Herein, we attempted to determine how daily LIPUS stimulation might affect the cell form and bone matrix protein expression in spheroids of human alveolar bone periosteal cells. Histological (toluidine blue) and immunohistochemical analyses (osteopontin (OP) and osteocalcin (OC)) of spheroids were conducted after 1, 3, 7, and 14 days of culture with daily LIPUS stimulation. Layered structures, distinguished by the directionality of the cells, were observed in the spheroids from day 1, with the layered structures becoming indistinct over time in the order of the irradiated side, non-irradiated side, and control. Immunohistochemical reactions indicating OP and OC production were observed in the order of the core, middle layer, and outer layer of the spheroids. Furthermore, positive reactions of OC and OP were seen over time in the order of the irradiation side, non-irradiation side, and controls. In addition, funicular structures were observed in the core from the outer layer of the spheroids on the irradiated side from day 1. These findings show that LIPUS irradiation promotes cell differentiation and changes the cell direction within spheroids. Nihon Shishubyo Gakkai Kaishi (J Jpn Soc Periodontol) 55(4):312-325, 2013.
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