University of Regensburg, Department of Orthodontics, Germany.
Private Practice, Curitiba, Brasil.
J Appl Oral Sci. 2023 May 29;31:e20220151. doi: 10.1590/1678-7757-2022-0151. eCollection 2023.
Many genes and signaling molecules are involved in orthodontic tooth movement, with mechanically and hypoxically stabilized HIF-1α having been shown to play a decisive role in periodontal ligament signaling during orthodontic tooth movement. Thus, this in vitro study aimed to investigate if genetic polymorphisms in HIF1A (Hypoxia-inducible factor α-subunits) influence the expression pattern of HIF-1α protein during simulated orthodontic compressive pressure.
Samples from human periodontal ligament fibroblasts were used and their DNA was genotyped using real time Polymerase chain reaction for the genetic polymorphisms rs2301113 and rs2057482 in HIF1A . For cell culture and protein expression experiments, six human periodontal ligament fibroblast cell lines were selected based on the patients' genotype. To simulate orthodontic compressive pressure in fibroblasts, a 2 g/cm2 force was applied under cell culture conditions for 48 hours. Protein expression was evaluated by Western Blot. Paired t-tests were used to compare HIF-1α expression with and without compressive pressure application and unpaired t-tests were used to compare expression between the genotypes in rs2057482 and rs2301113 (p<0.05).
The expression of HIF-1α protein was significantly enhanced by compressive pressure application regardless of the genotype (p<0.0001). The genotypes in the genetic polymorphisms rs2301113 and rs2057482 were not associated with HIF-1α protein expression (p>0.05).
Our study confirms that compressive pressure application enhances HIF-1α protein expression. We could not prove that the genetic polymorphisms in HIF1A affect HIF-1α protein expression by periodontal ligament fibroblasts during simulated orthodontic compressive force.
许多基因和信号分子参与正畸牙齿移动,机械和低氧稳定的 HIF-1α 已被证明在正畸牙齿移动期间牙周韧带信号传导中起决定性作用。因此,本体外研究旨在探讨 HIF1A(缺氧诱导因子α亚基)中的遗传多态性是否影响模拟正畸压缩压力下 HIF-1α 蛋白的表达模式。
使用来自人牙周韧带成纤维细胞的样本,并使用实时聚合酶链反应对 HIF1A 中的遗传多态性 rs2301113 和 rs2057482 进行基因分型。为了在成纤维细胞中模拟正畸压缩压力,在细胞培养条件下施加 2g/cm2 的力 48 小时。通过 Western Blot 评估蛋白质表达。使用配对 t 检验比较有和没有压缩压力应用时的 HIF-1α 表达,使用未配对 t 检验比较 rs2057482 和 rs2301113 中基因型之间的表达(p<0.05)。
无论基因型如何,压缩压力应用均可显著增强 HIF-1α 蛋白的表达(p<0.0001)。遗传多态性 rs2301113 和 rs2057482 中的基因型与 HIF-1α 蛋白表达无关(p>0.05)。
我们的研究证实,压缩压力应用可增强 HIF-1α 蛋白的表达。我们无法证明 HIF1A 中的遗传多态性会影响模拟正畸压力下牙周韧带成纤维细胞中 HIF-1α 蛋白的表达。