Redlich Meir, Asher Roos Hervé, Reichenberg Elisha, Zaks Batya, Mussig Dieter, Baumert Uwe, Golan Ilan, Palmon Aaron
Department of Orthodontics, Hadassah Faculty of Dental Medicine, Hebrew University, founded by the Alpha-Omega Fraternity, PO Box 12277, Jerusalem 91220, Israel.
Arch Oral Biol. 2004 Feb;49(2):119-24. doi: 10.1016/j.archoralbio.2003.08.002.
The ability of elastic fibers to respond to mechanical stimuli suggests that they play a central role in physiological adaptation to external stimuli including application of orthodontic force. The purpose of this study was to examine the effect of external pressure simulating orthodontic force on tropoelastin gene expression in cultured human periodontal ligament fibroblasts (hPDLF). External pressure simulation was achieved by centrifugation for 10, 30, 60, 90 and 120 min of hPDLF in a horizontal microplate rotor. Semi-quantitative RT-PCR analysis of tropoelastin mRNA was performed and beta-actin was used as an internal invariant control. While centrifugal force on mRNA levels of beta-actin showed almost no change, the mRNA levels of tropoelastin increased significantly to a peak level of more than four-fold after 30 min. Thereafter, at 60 min, the mRNA levels remained at more than three-fold. After 90 min, mRNA levels decreased to control levels. The finding that no changes in mRNA levels of beta-actin occurred during the first 90 min of centrifugation validates its use as an invariant control gene in such an experimental model. This study demonstrated that tropoelastin is expressed in hPDLF and that the pressure caused significant time-dependent upregulation of the tropoelastin gene. The responsiveness of the tropoelastin gene to force shows its possible clinical importance in orthodontic tooth movement. Further studies, however, are essential in order to learn whether the high expression of the gene in vitro will also be followed by corresponding protein synthesis and deposition in vivo in the extracellular matrix (ECM) of the periodontal ligament (PDL).
弹性纤维对机械刺激作出反应的能力表明,它们在对包括正畸力施加在内的外部刺激的生理适应中发挥着核心作用。本研究的目的是检测模拟正畸力的外部压力对培养的人牙周膜成纤维细胞(hPDLF)中原弹性蛋白基因表达的影响。通过在水平微孔板转子中对hPDLF进行10、30、60、90和120分钟的离心来实现外部压力模拟。对原弹性蛋白mRNA进行半定量RT-PCR分析,并使用β-肌动蛋白作为内部不变对照。虽然离心力对β-肌动蛋白mRNA水平几乎没有影响,但原弹性蛋白的mRNA水平在30分钟后显著增加至峰值水平,超过四倍。此后,在60分钟时,mRNA水平保持在三倍以上。90分钟后,mRNA水平降至对照水平。在离心的前90分钟内β-肌动蛋白mRNA水平没有变化这一发现,证实了其在这种实验模型中作为不变对照基因的用途。本研究表明,原弹性蛋白在hPDLF中表达,并且压力导致原弹性蛋白基因出现显著的时间依赖性上调。原弹性蛋白基因对力的反应性表明其在正畸牙齿移动中可能具有临床重要性。然而,为了了解该基因在体外的高表达是否也会伴随着相应的蛋白质合成以及在体内牙周膜(PDL)细胞外基质(ECM)中的沉积,还需要进一步的研究。