Knaup Isabel, Kramann Rafael, Sasula Martha-Julia, Mack Paula, Bastos Craveiro Rogério, Niederau Christian, Coenen Franziska, Neuss Sabine, Jankowski Joachim, Wolf Michael
Department of Orthodontics, Medical Faculty, RWTH Aachen University, Pauwelsstr. 30, 52074, Aachen, Germany.
Clinic for Renal and Hypertensive Disorders, Rheumatological and Immunological Diseases (Medical Clinic II), Medical Faculty, RWTH Aachen University, Aachen, Germany.
J Orofac Orthop. 2024 Aug 2. doi: 10.1007/s00056-024-00541-2.
To investigate the effect of tumor necrosis factor (TNF) on the growth of human periodontal ligament (PDL) cells, their osteogenic differentiation and modulation of their matrix secretion in vitro.
The influence of 10 ng/ml TNF on proliferation and metabolic activity of PDL cells was analyzed by cell counting (DAPI [4',6-diamidino-2-phenylindole] staining) and the MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay. In addition, cells were cultured under control conditions and osteogenic conditions (media containing 10 mM β-glycerophosphate). Quantitative expression analysis of genes encoding the osteogenic markers alkaline phosphatase (ALP), collagen type I alpha 1 chain (COL1A1), osteoprotegerin (OPG), and osteopontin (OPN) was performed after 7 and 14 days of cultivation. Calcium deposits were stained with alizarin red.
Our studies showed that 10 ng/ml TNF did not affect the survival and metabolic activity of PDL cells. Quantitative expression analysis revealed that long-term cultures with TNF impaired osteogenic cell fate at early and late developmental stages. Furthermore, TNF significantly reduced matrix secretion in PDL cells.
The present data confirm TNF as a regulatory factor of proinflammatory remodeling that influences the differentiation behavior but not the metabolism and cell proliferation of the periodontium. Therefore, TNF represents an interesting target for the regulation of orthodontic remodeling processes in the periodontium.
研究肿瘤坏死因子（TNF）对人牙周膜（PDL）细胞生长、成骨分化及其体外基质分泌调节的影响。
通过细胞计数（4',6-二脒基-2-苯基吲哚[DAPI]染色）和MTS（3-(4,5-二甲基噻唑-2-基)-5-(3-羧甲氧基苯基)-2-(4-磺基苯基)-2H-四唑）试验分析10 ng/ml TNF对PDL细胞增殖和代谢活性的影响。此外，细胞在对照条件和成骨条件（含10 mM β-甘油磷酸的培养基）下培养。培养7天和14天后，对编码成骨标志物碱性磷酸酶（ALP）、I型胶原α1链（COL1A1）、骨保护素（OPG）和骨桥蛋白（OPN）的基因进行定量表达分析。用茜素红对钙沉积进行染色。
我们的研究表明，10 ng/ml TNF不影响PDL细胞的存活和代谢活性。定量表达分析显示，长期用TNF培养会损害成骨细胞在早期和晚期发育阶段的命运。此外，TNF显著减少PDL细胞中的基质分泌。
目前的数据证实TNF是促炎重塑的调节因子，它影响牙周组织的分化行为，但不影响其代谢和细胞增殖。因此，TNF是调节牙周正畸重塑过程的一个有趣靶点。
