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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

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Abstract

Objectives

The aim of this study was to evaluate the combined effects of recombinant human bone morphogenetic protein - 9 (rhBMP-9) loaded onto absorbable collagen sponges (ACS) and low-intensity pulsed ultrasound (LIPUS) on bone formation in rat calvarial defects.

Materials and methods

Circular calvarial defects were surgically created in 18 Wistar rats, which were divided into LIPUS-applied (+) and LIPUS-non-applied (−) groups. The 36 defects in each group received ACS implantation (ACS group), ACS with rhBMP-9 (rhBMP-9/ACS group), or surgical control (control group), yielding the following six groups: ACS (+/−), rhBMP-9/ACS (+/−), and control (+/−). The LIPUS-applied groups received daily LIPUS exposure starting immediately after surgery. At 4 weeks, animals were sacrificed and their defects were investigated histologically and by microcomputed tomography.

Results

Postoperative clinical healing was uneventful at all sites. More new bone was observed in the LIPUS-applied groups compared with the LIPUS-non-applied groups. Newly formed bone area (NBA)/total defect area (TA) in the ACS (+) group (46.49 ± 7.56%) was significantly greater than that observed in the ACS (−) (34.31 ± 5.68%) and control (−) (31.13 ± 6.74%) groups (p < 0.05). The rhBMP-9/ACS (+) group exhibited significantly greater bone volume, NBA, and NBA/TA than the rhBMP-9/ACS (−) group (2.46 ± 0.65 mm3 vs. 1.76 ± 0.44 mm3, 1.25 ± 0.31 mm2 vs. 0.88 ± 0.22 mm2, and 62.80 ± 11.87% vs. 42.66 ± 7.03%, respectively) (p < 0.05). Furthermore, the rhBMP-9/ ACS (+) group showed the highest level of bone formation among all groups.

Conclusion

Within their limits, it can be concluded that LIPUS had osteopromotive potential and enhanced rhBMP-9-induced bone formation in calvarial defects of rats.

Clinical relevance

The use of rhBMP-9 with LIPUS stimulation can be a potential bone regenerative therapy for craniofacial/peri-implant bone defects.

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Acknowledgements

We thank Edanz Group (https://en-author-services.edanzgroup.com/ac) for editing a draft of this manuscript.

Funding

This study was supported by Grants-in-Aid for Scientific Research C (No. 19 K10169) and Grant-in-Aid for Young Scientists (No. 18 K17089) from the Japan Society for the Promotion of Science.

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Authors and Affiliations

  1. Department of Periodontology, Kagoshima University Graduate School of Medical and Dental Sciences, 8-35-1 Sakuragaoka, Kagoshima, 890-8544, Japan

    Takatomo Imafuji, Yoshinori Shirakata, Yukiya Shinohara, Toshiaki Nakamura & Kazuyuki Noguchi

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  1. Takatomo Imafuji
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  2. Yoshinori Shirakata
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  3. Yukiya Shinohara
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Correspondence to Yoshinori Shirakata.

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The experimental protocol was reviewed and approved by the ethical committee of the Animal Research Center of Kagoshima University, Japan (Approval No. D19011). This study conformed to the ARRIVE guidelines for preclinical animal studies.

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Imafuji, T., Shirakata, Y., Shinohara, Y. et al. 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 Invest 25, 5917–5927 (2021). https://doi.org/10.1007/s00784-021-03897-6

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