Abe Takaharu, Sumi Keisuke, Kunimatsu Ryo, Oki Nanae, Tsuka Yuji, Nakajima Kengo, Ando Kazuyo, Tanimoto Kotaro
Department of Orthodontics, Division of Oral Health and Development, Hiroshima University Hospital.
Department of Orthodontics, Applied Life Sciences, Hiroshima University Institute of Biomedical & Health Sciences.
J Oral Sci. 2019 Mar 28;61(1):30-35. doi: 10.2334/josnusd.17-0315. Epub 2018 Dec 11.
Transplantation of mesenchymal stem cells (MSCs) has been extensively studied in the field of regenerative medicine. Bone regeneration is achieved via the interaction of osteoblasts and osteoclasts. However, the influence of MSCs on osteoclasts is unknown. The purpose of this study was to investigate the effect of MSCs on the expression of genes for osteoclast differentiation factors using qPCR after indirect co-culture of MSCs and RAW264 cells. The numbers of osteoclasts after addition of soluble receptor activator of nuclear factor kappa B (NF-κB) ligand (sRANKL) were also compared. Expression of osteoprotegerin (OPG) by MSCs was significantly elevated in co-culture over time. The differentiation of RAW264 cells into mature osteoclasts following addition of sRANKL was significantly inhibited by co-culture with MSCs. Expression of RANK, colony stimulating factor 1 receptor, NF-κB, and nuclear factor of activated T-cell cytoplasmic 1 in RAW264 cells was significantly inhibited by co-culture with MSCs. Expression of OPG protein was higher in co-culture with RAW264 cells than in MSCs alone, and the expression level was clearly higher than that of RANKL. MSCs appeared to inhibit osteoclast differentiation via expression of OPG.
间充质干细胞（MSCs）移植已在再生医学领域得到广泛研究。骨再生是通过成骨细胞和破骨细胞的相互作用实现的。然而，MSCs对破骨细胞的影响尚不清楚。本研究的目的是在MSCs与RAW264细胞间接共培养后，使用qPCR研究MSCs对破骨细胞分化因子基因表达的影响。还比较了添加核因子κB受体活化因子配体（sRANKL）后破骨细胞的数量。随着共培养时间的延长，MSCs中骨保护素（OPG）的表达显著升高。与MSCs共培养显著抑制了添加sRANKL后RAW264细胞向成熟破骨细胞的分化。与MSCs共培养显著抑制了RAW264细胞中RANK、集落刺激因子1受体、核因子κB和活化T细胞胞质1核因子的表达。与RAW264细胞共培养时OPG蛋白的表达高于单独培养的MSCs，且表达水平明显高于RANKL。MSCs似乎通过OPG的表达抑制破骨细胞分化。
