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Received for publication January 3, 2006.
Revised March 14, 2006.
Accepted for publication March 15, 2006.
It has been shown that ultrasound (US) stimulation accelerates fracture healing in the animal models and in clinical studies. Here we found that US stimulation transiently increased the surface expression of 
2, 5, 
1 and 3 integrins in cultured osteoblasts, as shown by flow cytometric analysis and immuofluorescence staining. US stimulation increased prostaglandin E2 formation as well as the protein and mRNA levels of cyclooxygenase-2 (COX-2). At the mechanistic level, anti-integrin 51 and v3 antibodies or rhodostomin, a snake venom disintegrin, attenuated the US-induced COX-2 expression. Phosphatidylinositol 3-kinase (PI3K) inhibitors, LY294002 and wortmannin also inhibited the potentiating action of US. US stimulation increased the phosphorylation of focal adhesion kinase (FAK), extracellular signal-regulated kinases (ERK), p85 subunit of PI3K and serine 473 of Akt. COX-2 promoter activity was enhanced by US stimulation in cells transfected with pCOX2-Luc. Cotransfection with dominant negative mutant of FAK(Y397F), p85(
p85) , Akt(K179A) or ERK2(K52R) inhibited the potentiating action of US on COX-2 promoter activity. Expression of mineralized nodule was lower in dominant negative mutants of FAK, p85 and Akt-transfected clones than in vector-transfected control cells. Taken together, our results provide evidence that US stimulation increases COX-2 expression and promotes bone formation in osteoblasts via the integrin/FAK/PI3K/Akt and ERK signaling pathway.
Key words:
Protein Kinases (other), Cyclooxygenases, Tissue engineering
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