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Enhanced Bone Regeneration by Bone Morphogenetic Protein-2 after Pretreatment with Low-Intensity Pulsed Ultrasound in Distraction Osteogenesis

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Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

Background:

Bone morphogenetic protein 2 (BMP-2) and low-intensity pulsed ultrasound (LIPUS) have been used to enhance bone healing in distraction osteogenesis (DO). The aim of this study was to assess the synergistic effect of BMP-2 and LIPUS on bone regeneration in DO and to determine the optimal treatment strategy for enhanced bone regeneration.

Methods:

Rat mesenchymal stromal cells were treated with various application protocols of BMP-2 and LIPUS, and cell proliferation, alkaline phosphatase activity, and osteogenesis-related marker expression were evaluated. In vivo experiments were performed in a rabbit DO model according to the application protocols with different timings of BMP-2 and LIPUS application.

Results:

Application of BMP-2 after LIPUS pretreatment (BMP-2 after LIPUS) showed greater cell proliferation than LIPUS treatment alone, and higher ALP activity than all other treatment protocols. BMP-2 after LIPUS also exhibited increased gene expression levels of ALP, Cbfa1, and Osterix compared with LIPUS treatment alone. In vivo experiments revealed no significant differences in bone healing based on the timing of LIPUS treatment in DO. The combination of BMP-2 and LIPUS resulted in increased bone volume and bone mineral density compared with BMP-2 or LIPUS. Regarding the timing of BMP-2 application, the application of BMP-2 after LIPUS pretreatment led to greater bone volume than the application of BMP-2 before LIPUS.

Conclusion:

The results of this study suggest that the combined treatment of BMP-2 and LIPUS can lead to enhanced bone healing in DO and that effective bone healing can be achieved through the application of LIPUS before BMP-2.

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References

  1. Norholt SE, Jensen J, Schou S, Pedersen TK. Complications after mandibular distraction osteogenesis: a retrospective study of 131 patients. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2011;111:420–7.

    Article  Google Scholar 

  2. Swennen G, Schliephake H, Dempf R, Schierle H, Malevez C. Craniofacial distraction osteogenesis: a review of the literature: part 1: clinical studies. Int J Oral Maxillofac Surg. 2001;30:89–103.

    Article  CAS  Google Scholar 

  3. Master DL, Hanson PR, Gosain AK. Complications of mandibular distraction osteogenesis. J Craniofac Surg. 2010;21:1565–70.

    Article  Google Scholar 

  4. Hong P, Boyd D, Beyea SD, Bezuhly M. Enhancement of bone consolidation in mandibular distraction osteogenesis: a contemporary review of experimental studies involving adjuvant therapies. J Plast Reconstr Aesthet Surg. 2013;66:883–95.

    Article  Google Scholar 

  5. Makhdom AM, Hamdy RC. The role of growth factors on acceleration of bone regeneration during distraction osteogenesis. Tissue Eng Part B Rev. 2013;19:442–53.

    Article  CAS  Google Scholar 

  6. Xie LK, Wangrangsimakul K, Suttapreyasri S, Cheung LK, Nuntanaranont T. A preliminary study of the effect of low intensity pulsed ultrasound on new bone formation during mandibular distraction osteogenesis in rabbits. Int J Oral Maxillofac Surg. 2011;40:730–6.

    Article  CAS  Google Scholar 

  7. Rubin C, Bolander M, Ryaby JP, Hadjiargyrou M. The use of low-intensity ultrasound to accelerate the healing of fractures. J Bone Joint Surg Am. 2001;83:259–70.

    Article  CAS  Google Scholar 

  8. Hadjiargyrou M, McLeod K, Ryaby JP, Rubin C. Enhancement of fracture healing by low intensity ultrasound. Clin Orthop Relat Res. 1998;355:S216–29.

    Article  Google Scholar 

  9. Parvizi J, Parpura V, Greenleaf JF, Bolander ME. Calcium signaling is required for ultrasound-stimulated aggrecan synthesis by rat chondrocytes. J Orthop Res. 2002;20:51–7.

    Article  CAS  Google Scholar 

  10. Wang SJ, Lewallen DG, Bolander ME, Chao EY, Ilstrup DM, Greenleaf JF. Low intensity ultrasound treatment increases strength in a rat femoral fracture model. J Orthop Res. 1994;12:40–7.

    Article  CAS  Google Scholar 

  11. Yang KH, Parvizi J, Wang SJ, Lewallen DG, Kinnick RR, Greenleaf JF, et al. Exposure to low-intensity ultrasound increases aggrecan gene expression in a rat femur fracture model. J Orthop Res. 1996;14:802–9.

    Article  CAS  Google Scholar 

  12. El-Bialy TH, Royston TJ, Magin RL, Evans CA, Zaki Ael M, Frizzell LA. The effect of pulsed ultrasound on mandibular distraction. Ann Biomed Eng. 2002;30:1251–61.

    Article  Google Scholar 

  13. Schortinghuis J, Bronckers AL, Stegenga B, Raghoebar GM, de Bont LG. Ultrasound to stimulate early bone formation in a distraction gap: a double blind randomised clinical pilot trial in the edentulous mandible. Arch Oral Biol. 2005;50:411–20.

    Article  Google Scholar 

  14. Urist MR. Bone: formation by autoinduction. Science. 1965;150:893–9.

    Article  CAS  Google Scholar 

  15. Han JJ, Chang AR, Ahn J, Jung S, Hong J, Oh HK, et al. Efficacy and safety of rhBMP/beta-TCP in alveolar ridge preservation: a multicenter, randomized, open-label, comparative, investigator-blinded clinical trial. Maxillofac Plast Reconstr Surg. 2021;43:42.

    Article  Google Scholar 

  16. Herford AS, Boyne PJ. Reconstruction of mandibular continuity defects with bone morphogenetic protein-2 (rhBMP-2). J Oral Maxillofac Surg. 2008;66:616–24.

    Article  Google Scholar 

  17. Park JH, Kim JW, Kim SJ. Does the addition of bone morphogenetic protein 2 to platelet-rich fibrin improve healing after treatment for medication-related osteonecrosis of the jaw? J Oral Maxillofac Surg. 2017;75:1176–84.

    Article  Google Scholar 

  18. Cheung LK, Zheng LW. Effect of recombinant human bone morphogenetic protein-2 on mandibular distraction at different rates in an experimental model. J Craniofac Surg. 2006;17:100–8.

    Article  Google Scholar 

  19. Li G, Bouxsein ML, Luppen C, Li XJ, Wood M, Seeherman HJ, et al. Bone consolidation is enhanced by rhBMP-2 in a rabbit model of distraction osteogenesis. J Orthop Res. 2002;20:779–88.

    Article  CAS  Google Scholar 

  20. Sailhan F, Gleyzolle B, Parot R, Guerini H, Viguier E. Rh-BMP-2 in distraction osteogenesis: dose effect and premature consolidation. Injury. 2010;41:680–6.

    Article  Google Scholar 

  21. Angle SR, Sena K, Sumner DR, Virkus WW, Virdi AS. Combined use of low-intensity pulsed ultrasound and rhBMP-2 to enhance bone formation in a rat model of critical size defect. J Orthop Trauma. 2014;28:605–11.

    Article  Google Scholar 

  22. Sant’Anna EF, Leven RM, Virdi AS, Sumner DR. Effect of low intensity pulsed ultrasound and BMP-2 on rat bone marrow stromal cell gene expression. J Orthop Res. 2005;23:646–52.

    Article  CAS  Google Scholar 

  23. Wijdicks CA, Virdi AS, Sena K, Sumner DR, Leven RM. Ultrasound enhances recombinant human BMP-2 induced ectopic bone formation in a rat model. Ultrasound Med Biol. 2009;35:1629–37.

    Article  Google Scholar 

  24. Sakurakichi K, Tsuchiya H, Uehara K, Yamashiro T, Tomita K, Azuma Y. Effects of timing of low-intensity pulsed ultrasound on distraction osteogenesis. J Orthop Res. 2004;22:395–403.

    Article  Google Scholar 

  25. Mayr E, Laule A, Suger G, Ruter A, Claes L. Radiographic results of callus distraction aided by pulsed low-intensity ultrasound. J Orthop Trauma. 2001;15:407–14.

    Article  CAS  Google Scholar 

  26. Shimazaki A, Inui K, Azuma Y, Nishimura N, Yamano Y. Low-intensity pulsed ultrasound accelerates bone maturation in distraction osteogenesis in rabbits. J Bone Joint Surg Br. 2000;82:1077–82.

    Article  CAS  Google Scholar 

  27. Lai CH, Chen SC, Chiu LH, Yang CB, Tsai YH, Zuo CS, et al. Effects of low-intensity pulsed ultrasound, dexamethasone/TGF-beta1 and/or BMP-2 on the transcriptional expression of genes in human mesenchymal stem cells: chondrogenic vs. osteogenic differentiation. Ultrasound Med Biol. 2010;36:1022–33.

    Article  Google Scholar 

  28. Lou J, Xu F, Merkel K, Manske P. Gene therapy: adenovirus-mediated human bone morphogenetic protein-2 gene transfer induces mesenchymal progenitor cell proliferation and differentiation in vitro and bone formation in vivo. J Orthop Res. 1999;17:43–50.

    Article  CAS  Google Scholar 

  29. Selvamurugan N, Kwok S, Vasilov A, Jefcoat SC, Partridge NC. Effects of BMP-2 and pulsed electromagnetic field (PEMF) on rat primary osteoblastic cell proliferation and gene expression. J Orthop Res. 2007;25:1213–20.

    Article  CAS  Google Scholar 

  30. Imafuji T, Shirakata Y, Shinohara Y, Nakamura T, Noguchi K. Enhanced bone formation of calvarial bone defects by low-intensity pulsed ultrasound and recombinant human bone morphogenetic protein-9: a preliminary experimental study in rats. Clin Oral Investig. 2021;25:5917–27.

    Article  Google Scholar 

  31. Yonezawa H, Harada K, Ikebe T, Shinohara M, Enomoto S. Effect of recombinant human bone morphogenetic protein-2 (rhBMP-2) on bone consolidation on distraction osteogenesis: a preliminary study in rabbit mandibles. J Craniomaxillofac Surg. 2006;34:270–6.

    Article  Google Scholar 

  32. Egermann M, Lill CA, Griesbeck K, Evans CH, Robbins PD, Schneider E, et al. Effect of BMP-2 gene transfer on bone healing in sheep. Gene Ther. 2006;13:1290–9.

    Article  CAS  Google Scholar 

  33. Freitas RM, Spin-Neto R, Marcantonio Junior E, Pereira LA, Wikesjo UM, Susin C. Alveolar ridge and maxillary sinus augmentation using rhBMP-2: a systematic review. Clin Implant Dent Relat Res. 2015;17:e192-201.

    Article  Google Scholar 

  34. Gao TJ, Lindholm TS, Kommonen B, Ragni P, Paronzini A, Lindholm TC, et al. The use of a coral composite implant containing bone morphogenetic protein to repair a segmental tibial defect in sheep. Int Orthop. 1997;21:194–200.

    Article  CAS  Google Scholar 

  35. Viljanen VV, Gao TJ, Lindholm TC, Lindholm TS, Kommonen B. Xenogeneic moose (Alces alces) bone morphogenetic protein (mBMP)-induced repair of critical-size skull defects in sheep. Int J Oral Maxillofac Surg. 1996;25:217–22.

    Article  CAS  Google Scholar 

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Acknowledgements

This study was supported by a grant (No. 10045651) from the Advanced Technology Center R&D Project, Ministry of Trade, Industry and Energy, Republic of Korea and National Research Foundation of Korea (Grant No. NRF-2018R1A5A2024418). We thank Tae-Hyung Cho, and Beom Seok Lee for providing technical assistance with animal experiments.

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Authors and Affiliations

  1. Department of Oral and Maxillofacial Surgery, School of Dentistry, Seoul National University, Seoul, 03080, Republic of Korea

    Jeong Joon Han & Hoon Joo Yang

  2. Dental Research Institute, Seoul National University, Seoul, 03080, Republic of Korea

    Jeong Joon Han, Hoon Joo Yang & Soon Jung Hwang

  3. Hwang Soon Jung’s Dental Clinic for Oral and Maxillofacial Surgery, 349, Woonam Building 2, 3F, Gangnam-daero, Seocho-gu, Seoul, 06626, Republic of Korea

    Soon Jung Hwang

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  1. Jeong Joon Han
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Correspondence to Soon Jung Hwang.

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The animal studies were performed after receiving approval from the Institutional Animal Care and Use Committee (IACUC) of Seoul National University (IACUC no. SNU-141120-2).

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Han, J.J., Yang, H.J. & Hwang, S.J. Enhanced Bone Regeneration by Bone Morphogenetic Protein-2 after Pretreatment with Low-Intensity Pulsed Ultrasound in Distraction Osteogenesis. Tissue Eng Regen Med 19, 871–886 (2022). https://doi.org/10.1007/s13770-022-00457-1

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  • DOI: https://doi.org/10.1007/s13770-022-00457-1

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