title: Effect of low- and high-intensity pulsed ultrasound on collagen post-translational modifications in MC3T3-E1 osteoblasts.
abstract: Different intensities of pulsed ultrasound have distinct biological effects on bone mineralization in the process of bone fracture repair, even across a narrow range (e.g., 30-120 mW/cm(2)). The aim of our study was to elucidate the effect of low-intensity (30 mW/cm(2)) and high-intensity (120 mW/cm(2)) pulsed ultrasound on collagen metabolism by using MC3T3-E1 osteoblasts. Of special interest was the relationship between posttranslational collagen quality and prostaglandin E(2) activity. Cells with or without a cyclooxygenase-2 inhibitor, NS398, were exposed every day for four consecutive days to high-level or low-level intensities of pulsed ultrasound. We examined the, expression patterns of cyclooxygenase-2, lysyl oxidase, telopeptidyl lysyl hydroxylase (TLH), and helical lysyl hydroxylase by real-time polymerase chain reaction analysis. Quantitative analyses of reducible immature and nonreducible mature cross-links were also performed. Ultrasound at 30 mW/cm(2) upregulated TLH messenger RNA (mRNA) expression and enzyme activity compared to the control and resulted in increased relative concentrations of telopeptidyl hydroxylysine-derived cross-links. In addition to upregulated lysyl oxidase mRNA expression, increased total reducible and nonreducible cross-links were observed by 30 mW/cm(2) exposure compared to the control. In contrast, ultrasound at 120 mW/cm(2) had no obvious effect on collagen metabolism owing to high levels of endogenous prostaglandin E(2) induced by ultrasound. Our results showed that (1) low-intensity, but not high-intensity, ultrasound may accelerate the formation of the unique molecular packing of collagen fibers conducive to bone mineralization and that (2) the high dose of endogenous prostaglandin E(2) induced by pulsed ultrasound may be detrimental to calcifiable cross-link formation.
authors.affiliation: Department of Orthopaedic Surgery, Jikei University School of Medicine, 3-25-8, Nishi-Skinbaski, Minato-ku, Tokyo, 105-8461, Japan. xlink67@gol.com
authors.forename: M
authors.initials: M
authors.lastname: Saito
authors.forename: K
authors.initials: K
authors.lastname: Fujii
authors.forename: T
authors.initials: T
authors.lastname: Tanaka
authors.forename: S
authors.initials: S
authors.lastname: Soshi
mesh_terms: D016475:3T3 Cells; D000818:Animals; D003094:Collagen / Q000235:genetics / Q000378:metabolism*; D003432:Cross-Linking Reagents / Q000378:metabolism; D003470:Culture Media / Q000032:analysis; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D015232:Dinoprostone / Q000096:biosynthesis; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D008565:Membrane Proteins; D051379:Mice; D009578:Nitrobenzenes / Q000494:pharmacology; D010006:Osteoblasts / Q000000981:diagnostic imaging* / Q000378:metabolism; D008240:Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase / Q000378:metabolism; D011451:Prostaglandin-Endoperoxide Synthases / Q000187:drug effects; D011499:Protein Processing, Post-Translational; D008249:Protein-Lysine 6-Oxidase / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D013449:Sulfonamides / Q000494:pharmacology; D013997:Time Factors; D014465:Ultrasonics; D018617:Ultrasonography, Doppler, Pulsed / Q000379:methods*; D015854:Up-Regulation
chemical_list: D003432:Cross-Linking Reagents; D003470:Culture Media; D004791:Enzyme Inhibitors; D008565:Membrane Proteins; D009578:Nitrobenzenes; D012333:RNA, Messenger; D013449:Sulfonamides; C080955:N-(2-cyclohexyloxy-4-nitrophenyl)methanesulfonamide; D003094:Collagen; D008240:Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases; C496512:Ptgs1 protein, mouse; D008249:Protein-Lysine 6-Oxidase; D015232:Dinoprostone