Torii Daisuke, Konishi Kiyoshi, Watanabe Nobuyuki, Goto Shinichi, Tsutsui Takeki
Department of Pharmacology, The Nippon Dental University School of Life Dentistry at Tokyo, Tokyo, Japan.
Odontology. 2015 Jan;103(1):27-35. doi: 10.1007/s10266-013-0145-y. Epub 2014 Jan 8.
The periodontal ligament (PDL) consists of a group of specialized connective tissue fibers embedded in the alveolar bone and cementum that are believed to contain progenitors for mineralized tissue-forming cell lineages. These progenitors may contribute to regenerative cell therapy or tissue engineering methods aimed at recovery of tissue formation and functions lost in periodontal degenerative changes. Some reports using immortal clonal cell lines of cementoblasts, which are cells containing mineralized tissue-forming cell lineages, have shown that their phenotypic alteration and gene expression are associated with mineralization. Immortal, multipotential PDL-derived cell lines may be useful biological tools for evaluating differentiation-inducing agents. In this study, we confirmed the gene expression and mineralization potential of primary and immortal human PDL cells and characterized their immunophenotype. Following incubation with mineralization induction medium containing β-glycerophosphate, ascorbic acid, and dexamethasone, normal human PDL (Pel) cells and an immortal derivative line (Pelt) cells showed higher levels of mineralization compared with cells grown in normal growth medium. Both cell types were positive for putative surface antigens of mesenchymal cells (CD44, CD73, CD90, and CD105). They were also positive for stage-specific embryonic antigen-3, a marker of multipotential stem cells. Furthermore, PDL cells expressed cementum attachment protein and cementum protein 1 when cultured with recombinant human bone morphogenetic protein-2 or -7. The results suggest that normal and immortal human PDL cells contain multipotential mesenchymal stem cells with cementogenic potential.
牙周韧带（PDL）由一组嵌入牙槽骨和牙骨质的特殊结缔组织纤维组成，这些纤维被认为含有矿化组织形成细胞谱系的祖细胞。这些祖细胞可能有助于再生细胞治疗或组织工程方法，旨在恢复牙周退行性改变中丧失的组织形成和功能。一些使用成牙骨质细胞的永生克隆细胞系（这些细胞含有矿化组织形成细胞谱系）的报告表明，它们的表型改变和基因表达与矿化有关。永生的、多能的源自牙周韧带的细胞系可能是评估分化诱导剂的有用生物学工具。在本研究中，我们证实了原代和永生的人牙周韧带细胞的基因表达和矿化潜力，并对其免疫表型进行了表征。在用含有β-甘油磷酸、抗坏血酸和地塞米松的矿化诱导培养基孵育后，正常人牙周韧带（Pel）细胞和永生衍生系（Pelt）细胞与在正常生长培养基中生长的细胞相比，矿化水平更高。两种细胞类型对间充质细胞的假定表面抗原（CD44、CD73、CD90和CD105）均呈阳性。它们对多能干细胞的标志物阶段特异性胚胎抗原-3也呈阳性。此外，当用重组人骨形态发生蛋白-2或-7培养时，牙周韧带细胞表达牙骨质附着蛋白和牙骨质蛋白1。结果表明，正常和永生的人牙周韧带细胞含有具有成牙骨质潜力的多能间充质干细胞。
