Division of Pediatric Dentistry, Department of Oral Growth and Development, School of Dentistry, Health Sciences University of Hokkaido, 1757 Kanazawa, Ishikari-Tobetsu, Hokkaido, 061-0293, Japan.
Division of Oral Medicine and Pathology, Department of Human Biology and Pathophysiology, School of Dentistry, Health Sciences University of Hokkaido, 1757 Kanazawa, Ishikari-Tobetsu, Hokkaido, 061-0293, Japan.
Sci Rep. 2022 Jan 10;12(1):382. doi: 10.1038/s41598-021-04091-0.
The epithelial cell rests of Malassez (ERM) are essential in preventing ankylosis between the alveolar bone and the tooth (dentoalveolar ankylosis). Despite extensive research, the mechanism by which ERM cells suppress ankylosis remains uncertain; perhaps its varied population is to reason. Therefore, in this study, eighteen unique clones of ERM (CRUDE) were isolated using the single-cell limiting dilution and designated as ERM 1-18. qRT-PCR, ELISA, and western blot analyses revealed that ERM-2 and -3 had the highest and lowest amelogenin expression, respectively. Mineralization of human periodontal ligament fibroblasts (HPDLF) was reduced in vitro co-culture with CRUDE ERM, ERM-2, and -3 cells, but recovered when an anti-amelogenin antibody was introduced. Transplanted rat molars grown in ERM-2 cell supernatants produced substantially less bone than those cultured in other cell supernatants; inhibition was rescued when an anti-amelogenin antibody was added to the supernatants. Anti-Osterix antibody staining was used to confirm the development of new bones. In addition, next-generation sequencing (NGS) data were analysed to discover genes related to the distinct roles of CRUDE ERM, ERM-2, and ERM-3. According to this study, amelogenin produced by ERM cells helps to prevent dentoalveolar ankylosis and maintain periodontal ligament (PDL) space, depending on their clonal diversity.
牙周膜上皮细胞剩余物（ERM）对于防止牙槽骨和牙齿之间的粘连（牙牙槽骨粘连）是必不可少的。尽管进行了广泛的研究，但 ERM 细胞抑制粘连的机制仍不清楚；也许其多样化的群体是原因之一。因此，在这项研究中，使用单细胞有限稀释法分离了 18 个独特的 ERM 克隆（CRUDE），并将其命名为 ERM 1-18。qRT-PCR、ELISA 和 Western blot 分析显示，ERM-2 和 -3 的牙本质蛋白表达水平最高和最低。体外共培养 CRUDE ERM、ERM-2 和 -3 细胞可减少人牙周膜成纤维细胞（HPDLF）的矿化，但引入抗牙本质蛋白抗体后可恢复矿化。在 ERM-2 细胞上清液中培养的移植大鼠磨牙产生的骨量明显少于在其他细胞上清液中培养的磨牙；当在上清液中加入抗牙本质蛋白抗体时，抑制作用得到挽救。使用抗 Osterix 抗体染色来确认新骨的形成。此外，还分析了下一代测序（NGS）数据，以发现与 CRUDE ERM、ERM-2 和 ERM-3 不同作用相关的基因。根据这项研究，ERM 细胞产生的牙本质蛋白有助于防止牙牙槽骨粘连，并维持牙周膜（PDL）空间，这取决于它们的克隆多样性。
