International Journal of Stem Cells

Korean Society for Stem Cell Research (한국줄기세포학회)
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Low-Intensity Pulsed Ultrasound Promotes BMP9 Induced Osteoblastic Differentiation in Rat Dedifferentiated Fat Cells

  • Fumiaki Setoguchi (Department of Periodontology, Graduate School of Medical and Dental Sciences, Kagoshima University) ;
  • Kotaro Sena (Department of Periodontology, Graduate School of Medical and Dental Sciences, Kagoshima University) ;
  • Kazuyuki Noguchi (Department of Periodontology, Graduate School of Medical and Dental Sciences, Kagoshima University)
  • Received : 2023.03.11
  • Accepted : 2023.05.25
  • Published : 2023.11.30
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Abstract

Dedifferentiated fat cells (DFATs) isolated from mature adipocytes have a multilineage differentiation capacity similar to mesenchymal stem cells and are considered as promising source of cells for tissue engineering. Bone morphogenetic protein 9 (BMP9) and low-intensity pulsed ultrasound (LIPUS) have been reported to stimulate bone formation both in vitro and in vivo. However, the combined effect of BMP9 and LIPUS on osteoblastic differentiation of DFATs has not been studied. After preparing DFATs from mature adipose tissue from rats, DFATs were treated with different doses of BMP9 and/or LIPUS. The effects on osteoblastic differentiation were assessed by changes in alkaline phosphatase (ALP) activity, mineralization/calcium deposition, and expression of bone related genes; Runx2, osterix, osteopontin. No significant differences for ALP activity, mineralization deposition, as well as expression for bone related genes were observed by LIPUS treatment alone while treatment with BMP9 induced osteoblastic differentiation of DFATs in a dose dependent manner. Further, co-treatment with BMP9 and LIPUS significantly increased osteoblastic differentiation of DFATs compared to those treated with BMP9 alone. In addition, upregulation for BMP9-receptor genes was observed by LIPUS treatment. Indomethacin, an inhibitor of prostaglandin synthesis, significantly inhibited the synergistic effect of BMP9 and LIPUS co-stimulation on osteoblastic differentiation of DFATs. LIPUS promotes BMP9 induced osteoblastic differentiation of DFATs in vitro and prostaglandins may be involved in this mechanism.

Keywords

Acknowledgement

This work was supported by JSPS KAKENHI, Grant-in-Aid for Scientific Research (C), Grant Numbers JP18K09639, JP19K10169, JP21K09954, JP22K10039. The LIPUS equipment was provided by Teijin Pharma (Tokyo, Japan). We thank Dr. Yuko Mikuni-Takagaki (Kanagawa Dental Collage) and Mr. Yasuki Hanaoka (Teijin Pharma) for their support.

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