Exosomes from dental pulp cells attenuate bone loss in mouse experimental periodontitis
Exosomes from dental pulp cells attenuate bone loss in mouse experimental periodontitis
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DOI：
10.1111/jre.12949
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发表时间：
2021-11-26
影响因子：
3.5
通讯作者：
Tezuka, Ken-ichi
中科院分区：
文献类型：
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作者：
Shimizu, Yuta;Takeda-Kawaguchi, Tomoko;Tezuka, Ken-ichi
背景与目的外泌体是由多种细胞分泌的小泡状体。它们的生物效应在很大程度上取决于它们的细胞来源和起源细胞的生理状态。由间充质干细胞分泌的外泌体对多种疾病发挥治疗作用，并可作为干细胞疗法的潜在替代方案。我们以前建立和特点的人类白细胞抗原（HLA）单倍型同型（HHH）牙髓细胞（DPC）线从人类智齿。在这项研究中，我们的目的是研究局部施用HHH-DPC外泌体在牙周炎小鼠模型中的作用。方法从HHH-DPC中纯化Exosomes，进行粒径分析，并通过western blotting证实exosomes标记物的表达。我们还证实了外泌体对HHH-DPC和小鼠成骨细胞MC 3 T3-E1细胞迁移的影响。使用小鼠实验性牙周炎模型来评价exosomes在体内的作用。牙槽骨的形态通过显微计算机断层扫描（mu CT）和组织学分析进行评估。使用共培养系统评价外泌体对破骨细胞生成的影响。结果从HHH-DPC中纯化的exosomes具有均一的球形膜结构。HHH-DPC外泌体促进人DPC和小鼠成骨细胞的迁移。MTT法显示对人DPC的增殖有积极作用，但对小鼠成骨细胞无影响。用HHH-DPC外泌体处理不改变成骨细胞的分化。mu CT成像显示，外泌体抑制牙周炎小鼠模型中的牙槽骨吸收。虽然在外来体处理后脱钙组织切片中TRAP阳性破骨细胞样细胞的优势没有明显变化，但HHH-DPC外来体在体外显著抑制破骨细胞形成。结论HHH-DPC exosomes能促进人牙本质细胞和小鼠成骨细胞的迁移，有效地减轻牙周炎引起的骨丢失。
Background and objective Exosomes are small vesicles secreted from many cell types. Their biological effects largely depend on their cellular origin and the physiological state of the originating cells. Exosomes secreted by mesenchymal stem cells exert therapeutic effects against multiple diseases and may serve as potential alternatives to stem cell therapies. We previously established and characterized human leukocyte antigen (HLA) haplotype homo (HHH) dental pulp cell (DPC) lines from human wisdom teeth. In this study, we aimed to investigate the effect of local administration of HHH-DPC exosomes in a mouse model of periodontitis. Methods Exosomes purified from HHH-DPCs were subjected to particle size analysis, and expression of exosome markers was confirmed by western blotting. We also confirmed the effect of exosomes on the migration of both HHH-DPCs and mouse osteoblastic MC3T3-E1 cells. A mouse experimental periodontitis model was used to evaluate the effect of exosomes in vivo. The morphology of alveolar bone was assessed by micro-computed tomography (mu CT) and histological analysis. The effect of exosomes on osteoclastogenesis was evaluated using a co-culture system. Results The exosomes purified from HHH-DPCs were homogeneous and had a spherical membrane structure. HHH-DPC exosomes promoted the migration of both human DPCs and mouse osteoblastic cells. The MTT assay showed a positive effect on the proliferation of human DPCs, but not on mouse osteoblastic cells. Treatment with HHH-DPC exosomes did not alter the differentiation of osteoblastic cells. Imaging with mu CT revealed that the exosomes suppressed alveolar bone resorption in the mouse model of periodontitis. Although no change was apparent in the dominance of TRAP-positive osteoclast-like cells in decalcified tissue sections upon exosome treatment, HHH-DPC exosomes significantly suppressed osteoclast formation in vitro. Conclusions HHH-DPC exosomes stimulated the migration of human DPCs and mouse osteoblastic cells and effectively attenuated bone loss due to periodontitis.