Ultrasound stimulates nitric oxide and prostaglandin E2 production by human osteoblasts

Abstract

We have previously shown that the therapeutic range of ultrasound heals osteoradionecrotic bone and induces bone formation in vitro. It is well established that nitric oxide (NO) and prostaglandins are crucial early mediators in mechanically induced bone formation. The therapeutic range of ultrasound may act in the same way; therefore, we have investigated the effect of the therapeutic range of ultrasound on NO induction and prostaglandin E2 (PGE2) production in vitro. Two ultrasound machines were evaluated, "traditional" (1 MHz, pulsed 1:4, tested at four intensities) and a "long-wave" (45 kHz, continuous, also tested at four intensities) devices. Ultrasound was applied to human mandibular osteoblasts for 5 min, and incubated at 37°C for up to 24 h. The control group (sham insonated) was treated in the same way. NO was determined by measuring the nitrite concentration in the culture media colorimetrically, and PGE2 was assayed by radioimmunoassay. Ultrasound produced a significant increase in both induced nitrite and PGE2 production. The NO synthesis appeared to be via inducible NO synthase (iNOS) on the basis of the time course and levels of nitrite obtained, although the inhibition of other NOS isoforms by aminoguanidine cannot be excluded. PGE2 synthesis appeared to be via COX-2. With the 45 kHz machine, a significant increase in NO was achieved at three intensities, 5, 30, and 50 mW/cm2. The 1 MHz machine stimulated the synthesis of both NO and PGE2, but was significant at only one dose (0.1 W/cm2(SAPA)). There was no difference between the two machines with regard to PGE2 synthesis. The time-course experiment revealed peak production to be 12-18 h for both NO and PGE2. The therapeutic range of ultrasound stimulates both NO and PGE2 synthesis by human osteoblasts, and the 45 kHz machine appeared to be more effective than the traditional short-wave length. These results may reflect the healing effect of ultrasound on fractures and osteoradionecrosis. © 2002 by Elsevier Science Inc. All rights reserved.