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

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.