The present study was designed to determine the underlying mechanism of low-intensity pulsed ultrasound (LIPUS) induced alveolar bone remodeling and the role of BMP-2 expression in a rat orthodontic tooth movement model. Orthodontic appliances were placed between the homonymy upper first molars and the upper central incisors in rats under general anesthesia, followed by daily 20-min LIPUS or sham LIPUS treatment beginning at day 0. Tooth movement distances and molecular changes were evaluated at each observation point. In vitro and in vivo studies were conducted to detect HGF (Hepatocyte growth factor)/Runx2/BMP-2 signaling pathways and receptor activator of NFκB ligand (RANKL) expression by quantitative real time PCR (qRT-PCR), Western blot and immunohistochemistry. At day 3, LIPUS had no effect on the rat orthodontic tooth movement distance and BMP-2-induced alveolar bone remodeling. However, beginning at day 5 and for the following time points, LIPUS significantly increased orthodontic tooth movement distance and BMP-2 signaling pathway and RANKL expression compared with the control group. The qRT-PCR and Western blot data in vitro and in vivo to study BMP-2 expression were consistent with the immunohistochemistry observations. The present study demonstrates that LIPUS promotes alveolar bone remodeling by stimulating the HGF/Runx2/BMP-2 signaling pathway and RANKL expression in a rat orthodontic tooth movement model, and LIPUS increased BMP-2 expression via Runx2 regulation.
References
[1]
Spadaro JA (1997) Mechanical and electrical interactions in bone remodeling. Bioelectromagnetics 18: 193–202.
[2]
Kobayashi Y, Takagi H, Sakai H, Hashimoto F, Mataki S, et al. (1998) Effects of local administration of osteocalcin on experimental tooth movement. Angle Orthod 68: 259–266.
[3]
Iino S, Sakoda S, Ito G, Nishimori T, Ikeda T, et al.. (2007) Acceleration of orthodontic tooth movement by alveolar corticotomy in the dog. Am J Orthod Dentofacial Orthop 131: 448 e441–448.
[4]
Kim SJ, Park YG, Kang SG (2009) Effects of Corticision on paradental remodeling in orthodontic tooth movement. Angle Orthod 79: 284–291.
[5]
Lee W, Karapetyan G, Moats R, Yamashita DD, Moon HB, et al. (2008) Corticotomy-/osteotomy-assisted tooth movement microCTs differ. J Dent Res 87: 861–867.
[6]
Iglesias-Linares A, Moreno-Fernandez AM, Yanez-Vico R, Mendoza-Mendoza A, Gonzalez-Moles M, et al. (2011) The use of gene therapy vs. corticotomy surgery in accelerating orthodontic tooth movement. Orthod Craniofac Res 14: 138–148.
[7]
Strippoli J, Aknin JJ (2012) [Accelerated tooth movement by alveolar corticotomy or piezocision]. Orthod Fr 83: 155–164.
[8]
Fujita S, Yamaguchi M, Utsunomiya T, Yamamoto H, Kasai K (2008) Low-energy laser stimulates tooth movement velocity via expression of RANK and RANKL. Orthod Craniofac Res 11: 143–155.
[9]
Hadjiargyrou M, McLeod K, Ryaby JP, Rubin C (1998) Enhancement of fracture healing by low intensity ultrasound. Clin Orthop Relat Res: S216–229.
[10]
Malizos KN, Papachristos AA, Protopappas VC, Fotiadis DI (2006) Transosseous application of low-intensity ultrasound for the enhancement and monitoring of fracture healing process in a sheep osteotomy model. Bone 38: 530–539.
[11]
Sun JS, Hong RC, Chang WH, Chen LT, Lin FH, et al. (2001) In vitro effects of low-intensity ultrasound stimulation on the bone cells. J Biomed Mater Res 57: 449–456.
[12]
Li JK, Chang WH, Lin JC, Ruaan RC, Liu HC, et al. (2003) Cytokine release from osteoblasts in response to ultrasound stimulation. Biomaterials 24: 2379–2385.
[13]
Sena K, Leven RM, Mazhar K, Sumner DR, Virdi AS (2005) Early gene response to low-intensity pulsed ultrasound in rat osteoblastic cells. Ultrasound Med Biol 31: 703–708.
[14]
Sant'Anna EF, Leven RM, Virdi AS, Sumner DR (2005) Effect of low intensity pulsed ultrasound and BMP-2 on rat bone marrow stromal cell gene expression. J Orthop Res 23: 646–652.
[15]
Wijdicks CA, Virdi AS, Sena K, Sumner DR, Leven RM (2009) Ultrasound enhances recombinant human BMP-2 induced ectopic bone formation in a rat model. Ultrasound Med Biol 35: 1629–1637.
[16]
Suzuki A, Takayama T, Suzuki N, Kojima T, Ota N, et al. (2009) Daily low-intensity pulsed ultrasound stimulates production of bone morphogenetic protein in ROS 17/2.8 cells. J Oral Sci 51: 29–36.
[17]
Chan CW, Qin L, Lee KM, Zhang M, Cheng JC, et al. (2006) Low intensity pulsed ultrasound accelerated bone remodeling during consolidation stage of distraction osteogenesis. J Orthop Res 24: 263–270.
[18]
Inubushi T, Tanaka E, Rego EB, Kitagawa M, Kawazoe A, et al. (2008) Effects of ultrasound on the proliferation and differentiation of cementoblast lineage cells. J Periodontol 79: 1984–1990.
[19]
Martinez de Albornoz P, Khanna A, Longo UG, Forriol F, Maffulli N (2011) The evidence of low-intensity pulsed ultrasound for in vitro, animal and human fracture healing. Br Med Bull 100: 39–57.
[20]
Coords M, Breitbart E, Paglia D, Kappy N, Gandhi A, et al. (2011) The effects of low-intensity pulsed ultrasound upon diabetic fracture healing. J Orthop Res 29: 181–188.
[21]
El-Bialy T, Hassan A, Albaghdadi T, Fouad HA, Maimani AR (2006) Growth modification of the mandible with ultrasound in baboons: a preliminary report. Am J Orthod Dentofacial Orthop 130: 435 e437–414.
[22]
El-Bialy T, El-Shamy I, Graber TM (2004) Repair of orthodontically induced root resorption by ultrasound in humans. Am J Orthod Dentofacial Orthop 126: 186–193.
[23]
Urist MR (1965) Bone: formation by autoinduction. Science 150: 893–899.
[24]
Wozney JM, Rosen V, Celeste AJ, Mitsock LM, Whitters MJ, et al. (1988) Novel regulators of bone formation: molecular clones and activities. Science 242: 1528–1534.
[25]
Rosen V (2009) BMP2 signaling in bone development and repair. Cytokine Growth Factor Rev 20: 475–480.
King GN, King N, Cruchley AT, Wozney JM, Hughes FJ (1997) Recombinant human bone morphogenetic protein-2 promotes wound healing in rat periodontal fenestration defects. J Dent Res 76: 1460–1470.
[28]
Takiguchi T, Kobayashi M, Nagashima C, Yamaguchi A, Nishihara T, et al. (1999) Effect of prostaglandin E2 on recombinant human bone morphogenetic protein-2-stimulated osteoblastic differentiation in human periodontal ligament cells. J Periodontal Res 34: 431–436.
[29]
Jung RE, Glauser R, Scharer P, Hammerle CH, Sailer HF, et al. (2003) Effect of rhBMP-2 on guided bone regeneration in humans. Clin Oral Implants Res 14: 556–568.
[30]
Grano M, Galimi F, Zambonin G, Colucci S, Cottone E, et al. (1996) Hepatocyte growth factor is a coupling factor for osteoclasts and osteoblasts in vitro. Proc Natl Acad Sci U S A 93: 7644–7648.
[31]
D'Ippolito G, Schiller PC, Perez-stable C, Balkan W, Roos BA, et al. (2002) Cooperative actions of hepatocyte growth factor and 1,25-dihydroxyvitamin D3 in osteoblastic differentiation of human vertebral bone marrow stromal cells. Bone 31: 269–275.
[32]
Tsai SY, Huang YL, Yang WH, Tang CH (2012) Hepatocyte growth factor-induced BMP-2 expression is mediated by c-Met receptor, FAK, JNK, Runx2, and p300 pathways in human osteoblasts. Int Immunopharmacol 13: 156–162.
[33]
Oshiro T, Shiotani A, Shibasaki Y, Sasaki T (2002) Osteoclast induction in periodontal tissue during experimental movement of incisors in osteoprotegerin-deficient mice. Anat Rec 266: 218–225.
[34]
Kanzaki H, Chiba M, Arai K, Takahashi I, Haruyama N, et al. (2006) Local RANKL gene transfer to the periodontal tissue accelerates orthodontic tooth movement. Gene Ther 13: 678–685.
[35]
Borsje MA, Ren Y, de Haan-Visser HW, Kuijer R Comparison of low-intensity pulsed ultrasound and pulsed electromagnetic field treatments on OPG and RANKL expression in human osteoblast-like cells. Angle Orthod 80: 498–503.
[36]
Kawasaki K, Shimizu N (2000) Effects of low-energy laser irradiation on bone remodeling during experimental tooth movement in rats. Lasers Surg Med 26: 282–291.
[37]
Kohno T, Matsumoto Y, Kanno Z, Warita H, Soma K (2002) Experimental tooth movement under light orthodontic forces: rates of tooth movement and changes of the periodontium. J Orthod 29: 129–135.
[38]
Fung CH, Cheung WH, Pounder NM, de Ana FJ, Harrison A, et al. (2012) Effects of different therapeutic ultrasound intensities on fracture healing in rats. Ultrasound Med Biol 38: 745–752.
[39]
Reher P, Harris M, Whiteman M, Hai HK, Meghji S (2002) Ultrasound stimulates nitric oxide and prostaglandin E2 production by human osteoblasts. Bone 31: 236–241.
[40]
Hayton MJ, Dillon JP, Glynn D, Curran JM, Gallagher JA, et al. (2005) Involvement of adenosine 5'-triphosphate in ultrasound-induced fracture repair. Ultrasound Med Biol 31: 1131–1138.
[41]
Wang Y, Li Y, Fan X, Zhang Y, Wu J, et al. (2011) Early proliferation alteration and differential gene expression in human periodontal ligament cells subjected to cyclic tensile stress. Arch Oral Biol 56: 177–186.
[42]
Shiraishi R, Masaki C, Toshinaga A, Okinaga T, Nishihara T, et al. (2011) The effects of low-intensity pulsed ultrasound exposure on gingival cells. J Periodontol 82: 1498–1503.
[43]
Cowin SC, Moss-Salentijn L, Moss ML (1991) Candidates for the mechanosensory system in bone. J Biomech Eng 113: 191–197.
[44]
Mitsui N, Suzuki N, Maeno M, Yanagisawa M, Koyama Y, et al. (2006) Optimal compressive force induces bone formation via increasing bone morphogenetic proteins production and decreasing their antagonists production by Saos-2 cells. Life Sci 78: 2697–2706.
[45]
Kim SJ, Moon SU, Kang SG, Park YG (2009) Effects of low-level laser therapy after Corticision on tooth movement and paradental remodeling. Lasers Surg Med 41: 524–533.
[46]
Azuma Y, Ito M, Harada Y, Takagi H, Ohta T, et al. (2001) Low-intensity pulsed ultrasound accelerates rat femoral fracture healing by acting on the various cellular reactions in the fracture callus. J Bone Miner Res 16: 671–680.
[47]
Katano M, Naruse K, Uchida K, Mikuni-Takagaki Y, Takaso M, et al. (2011) Low intensity pulsed ultrasound accelerates delayed healing process by reducing the time required for the completion of endochondral ossification in the aged mouse femur fracture model. Exp Anim 60: 385–395.
[48]
El-Bialy T, Lam B, Aldaghreer S, Sloan AJ (2011) The effect of low intensity pulsed ultrasound in a 3D ex vivo orthodontic model. J Dent 39: 693–699.
[49]
Hsieh PC, Kenagy RD, Mulvihill ER, Jeanette JP, Wang X, et al. (2006) Bone morphogenetic protein 4: potential regulator of shear stress-induced graft neointimal atrophy. J Vasc Surg 43: 150–158.
[50]
Suzuki A, Takayama T, Suzuki N, Sato M, Fukuda T, et al. (2009) Daily low-intensity pulsed ultrasound-mediated osteogenic differentiation in rat osteoblasts. Acta Biochim Biophys Sin (Shanghai) 41: 108–115.
[51]
Long MW, Robinson JA, Ashcraft EA, Mann KG (1995) Regulation of human bone marrow-derived osteoprogenitor cells by osteogenic growth factors. J Clin Invest 95: 881–887.
[52]
Khanna A, Nelmes RT, Gougoulias N, Maffulli N, Gray J (2009) The effects of LIPUS on soft-tissue healing: a review of literature. Br Med Bull 89: 169–182.
