Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago.
Pun Hlaing Hospitals.
J Oral Sci. 2021 Oct 1;63(4):341-346. doi: 10.2334/josnusd.21-0164. Epub 2021 Sep 15.
This study aimed to determine expressions of hepatocyte growth factor (HGF) and MET proto-oncogene receptor tyrosine kinase (MET) in palatal periosteum (PP) and to examine the effect of HGF/MET on osteogenic differentiation of human palatal periosteum-derived mesenchymal stem cells (PD-MSCs).
HGF/MET proteins in human palatal periosteum (n = 3) were localized using immunohistochemistry. PD-MSCs (n = 3) were cultured in serum-free Essential 8 (E8) medium or osteogenic medium with and without Capmatinib, a selective ATP-inhibitor of MET. HGF concentration in vitro was measured with ELISA. Relative gene expression was quantified from PD-MSCs by quantitative reverse transcription real-time polymerase chain reaction.
Immunohistochemistry detected co-localization of HGF and MET protein in PP. HGF protein levels were significantly higher (P < 0.05) in osteogenic media (day 21: 12.19 ± 8.36 ng/mL) than in E8 medium (day 21: 0.42 ± 0.72 ng/mL). MET inhibitor had a limited feedback effect on the expression profile of the osteogenic genes tested. Gene expression levels for all but three genes were comparable in serum-free and osteogenic media at all time points.
HGF/MET present in human PP and HGF is upregulated in vitro during osteogenesis; however the targeted pathways controlled by MET may not involve osteoblast maturation.
本研究旨在确定肝细胞生长因子（HGF）和 MET 原癌基因受体酪氨酸激酶（MET）在腭骨膜（PP）中的表达，并研究 HGF/MET 对人腭骨膜来源间充质干细胞（PD-MSCs）成骨分化的影响。
采用免疫组织化学法定位人腭骨膜（n=3）中的 HGF/MET 蛋白。在无血清基本 8 培养基（E8）或成骨培养基中培养 PD-MSCs（n=3），并加入 MET 的选择性 ATP 抑制剂卡马替尼（Capmatinib）。采用 ELISA 法检测体外 HGF 浓度。通过定量逆转录实时聚合酶链反应从 PD-MSCs 中定量相对基因表达。
免疫组织化学检测到 HGF 和 MET 蛋白在 PP 中的共定位。成骨培养基中 HGF 蛋白水平（第 21 天：12.19±8.36ng/mL）明显高于 E8 培养基（第 21 天：0.42±0.72ng/mL）（P<0.05）。MET 抑制剂对所测试的成骨基因表达谱仅有有限的反馈作用。在所有时间点，无血清培养基和成骨培养基中的所有基因表达水平除三个基因外均相似。
HGF/MET 存在于人 PP 中，体外成骨过程中 HGF 上调；然而，MET 控制的靶向途径可能不涉及成骨细胞成熟。
