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Combination of low-intensity pulsed ultrasound and C3H10T1/2 cells promotes bone-defect healing

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Abstract

Aims

We systematically investigated the effect of combined use of low-intensity pulsed ultrasound (LIPUS) and bone mesenchymal stem cells C3H10T1/2 on bone-defect healing.

Methods

C3H10T1/2 cells were first induced into a stationary phase by incubation with low fetal bovine serum (5 ml/l) for five days and then sonicated with LIPUS for ten minutes once every day for five consecutive days. The same LIPUS treatment combined with C3H10T1/2 cells, which were incubated in regular fetal bovine serum (10 ml/l) were used to aid femoral fracture healing in Sprague–Dawley rats during four consecutive weeks. C3H10T1/2 cell proliferation activity was detected by MTT assay. Cell-cycle changes were determined, and cell proliferation index was calculated using flow cytometry. Bone reparation was evaluated by X-ray imaging and hematoxylin and eosin (H&E) staining during the healing process.

Results

LIPUS promoted C3H10T1/2 cell proliferation, the mechanism of which was possibly the up-regulation of Bmi-1 gene expression. At the end of week two after combined use of LIPUS and C3H10T1/2, the femoral gap was reduced on X-ray images. According to H&E staining results, new bone had homogeneous and similar density compared with normal surrounding bone after combined use of LIPUS and C3H10T1/2. At the end of week four, bone defects could not be observed by X-ray in all four groups and repaired bone substance in all four groups could be observed by H&E staining.

Conclusions

LIPUS treatment effectively promotes C3H10T1/2 cells to enter the growth/split phase from the stationary phase. This process enhances cell proliferation, which consequently promotes bone-defect healing.

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Acknowledgments

This study was supported by the National Basic Research Program of China (No. 2011CB707900), the National Natural Science Fund by the Chinese National Science Foundation (No. 81127901, 31000435, and 11274404) and Scientific and Technological Research Program of Chongqing Municipal Education Commission (No. KJ130329).

Disclosures

All authors declare no competing interests.

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    Authors and Affiliations

    1. State Key Laboratory of Ultrasound Engineering in Medicine Co-Founded by Chongqing and the Ministry of Science and Technology, Chongqing Key Laboratory of Biomedical Engineering, College of Biomedical Engineering, Chongqing Medical University, No. 1 Yixueyuan Road, Chongqing, 400016, People’s Republic of China

      Ruixin He, Weichen Zhou, Yu Zhang, Shuai Hu, Haisheng Yu, Yueping Luo, Baoru Liu, Jianbo Ran, Yan Wang & Wenzhi Chen

    2. Department of Physics, University of Vermont, Burlington, VT, USA

      Junru Wu

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    1. Ruixin He
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    Correspondence to Yan Wang or Wenzhi Chen.

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    Ruixin He and Weichen Zhou contributed equally to this work.

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    He, R., Zhou, W., Zhang, Y. et al. Combination of low-intensity pulsed ultrasound and C3H10T1/2 cells promotes bone-defect healing. International Orthopaedics (SICOT) 39, 2181–2189 (2015). https://doi.org/10.1007/s00264-015-2898-0

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