Establishment of a Primary Culture System of Human Periodontal Ligament Cells that Differentiate into Cementum Protein 1-expressing Cementoblast-like Cells
Establishment of a Primary Culture System of Human Periodontal Ligament Cells that Differentiate into Cementum Protein 1-expressing Cementoblast-like Cells
复制标题
DOI：
10.21873/invivo.11480
复制
发表时间：
2019-03-01
期刊：
影响因子：
2.3
通讯作者：
Yokose, Satoshi
中科院分区：
文献类型：
--
作者：
Kadokura, Hiroshi;Yamazaki, Takahide;Yokose, Satoshi
背景/目的：更好地了解牙骨质形成和成牙骨质分化将有助于牙周治疗。本研究旨在建立反映牙周组织牙骨质形成的细胞培养体系，并确定分离培养的原代人牙周韧带（PDL）细胞是否可用于成牙骨质分化的研究。材料与方法：PDL细胞从人PDL组织片段外植体中分离得到。PDL细胞在含有/3-甘油磷酸和抗坏血酸的分化培养基中培养21天。用碱性磷酸酶（ALP）和von Kossa染色检测细胞的变化。实时聚合酶链反应也用于骨水泥蛋白1 (CEMP1)，这是骨水泥母细胞及其祖细胞的特异性标志物。结果：第5天，少量纤维状的PDL细胞ALP阳性。第7天，几乎所有细胞ALP均呈阳性。第14天出现矿化结节，von Kossa染色阳性；到第21天，结节的数量和大小逐渐增加。第5天检测到CEMP1的表达，第7天逐渐升高，第14天达到峰值，第21天下降。结论：利用人PDL细胞建立了反映牙骨质形成的培养体系。结果表明，该培养方法对骨水泥发生和成水泥细胞分化的研究具有方便和实用的意义。
Background/Aim: A better understanding of cementogenesis and cementoblast differentiation would be useful for periodontal therapy. The aim of this study was to establish a cell culture system that reflects cementum formation in periodontal tissue and determine whether or not isolated and cultured primary human periodontal ligament (PDL) cells could be used for the study of the differentiation of cementoblast. Materials and Methods: PDL cells were isolated from the outgrowths of tissue fragments of human PDL. PDL cells were incubated for up to 21 days in differentiation medium containing /3-glycerophosphate and ascorbic acid. The changes in the cells were detected by alkaline phosphatase (ALP) and von Kossa staining. Real-time polymerase chain reaction was also performed for cementum protein 1 (CEMP1), which is a specific marker of cementoblasts and their progenitors. Results: On day 5, a small number of PDL cells, which were fibrous, were positive for ALP. On day 7, almost all cells were positive for ALP. On day 14, mineralization nodules appeared, as seen by positive von Kossa staining; the nodules increased in number and size by day 21. The expression of CEMP1 was detected on day 5, and its expression level increased gradually by day 7, reached a peak on day 14, and decreased by day 21. Conclusion: Human PDL cells were used to establish a culture system that reflects cementum formation. Our results suggested that this culture method is convenient and useful for the study of cementogenesis and cementoblast differentiation.