Annals of Anatomy ( IF 2.0 ) Pub Date : 2021-01-02 , DOI: 10.1016/j.aanat.2020.151668 Irma Azraq 1 , Rogerio B Craveiro 1 , Christian Niederau 1 , Julia Brockhaus 1 , Asisa Bastian 1 , Isabel Knaup 1 , Sabine Neuss 2 , Michael Wolf 1
Cementoblasts, located on the tooth root surface covered with cementum, are considered to have tooth protecting abilities. They prevent tissue damage and secure teeth anchorage inside the periodontal ligament during mechanical stress. However, the involvement of cementoblasts in mechanical compression induced periodontal remodeling needs to be identified and better understood.
Here, we investigated the effect of static compressive stimulation, simulating the compression side of orthodontic force and cell confluence on a murine cementoblast cell line (OC/CM). The influence of cell confluence in cementoblast cells was analyzed by MTS assay and immunostaining. Furthermore, mRNA and protein expression were investigated by real-time RT-PCR and western blotting at different confluence grades and after mechanical stimulation.
We observed that cementoblast cell proliferation increases with increasing confluence grades, while cell viability decreases in parallel. Gene expression of remodeling markers is regulated by compressive force. In addition, cementoblast confluence plays a crucial role in this regulation. Confluent cementoblasts show a significantly higher basal expression of Bsp, Osterix, Alpl, Vegfa, Mmp9, Tlr2 and Tlr4 compared to sub-confluent cells. After compressive force of 48 h at 60% confluence, an upregulation of Bsp, Osterix, Alpl, Vegf and Mmp9 is observed. In contrast, at high confluence, all analyzed genes were downregulated through mechanical stress. We also proved a regulation of ERK, phospho-ERK and phospho-AKT dependent on compressive force. In summary, our findings provide evidence that cementoblast physiology and metabolism is highly regulated in a cell confluence-dependent manner and by mechanical stimulation.
中文翻译:
ERK 和 AKT 的基因表达和磷酸化取决于鼠成牙骨质细胞中的机械力和细胞融合度
位于被牙骨质覆盖的牙根表面的成牙骨质细胞被认为具有保护牙齿的能力。它们可防止组织损伤并在机械应力期间将牙齿固定在牙周韧带内。然而,需要识别和更好地理解成牙骨质细胞在机械压缩诱导的牙周重塑中的参与。
在这里,我们研究了静态压缩刺激的影响,模拟了正畸力的压缩侧和细胞汇合对小鼠成牙骨质细胞系 (OC/CM) 的影响。通过MTS测定和免疫染色分析成牙骨质细胞中细胞汇合的影响。此外,在不同汇合度和机械刺激后,通过实时 RT-PCR 和蛋白质印迹研究 mRNA 和蛋白质表达。
我们观察到成牙骨质细胞增殖随着汇合度的增加而增加,而细胞活力同时降低。重塑标记的基因表达受压缩力调节。此外,成牙骨质细胞汇合在这种调节中起着至关重要的作用。与亚融合细胞相比,融合的成牙骨质细胞显示出显着更高的Bsp、Osterix、Alpl、Vegfa、Mmp9、Tlr2和Tlr4 的基础表达。在 60% 汇合处加压 48 小时后,Bsp、Osterix、Alpl、Vegf和Mmp9 的上调被观察到。相比之下,在高汇合时,所有分析的基因都通过机械应力下调。我们还证明了依赖于压缩力的 ERK、磷酸化 ERK 和磷酸化 AKT 的调节。总之,我们的研究结果提供证据表明,成牙骨质细胞生理和代谢以细胞融合依赖性方式和机械刺激受到高度调节。