FGF1 对人牙周膜成纤维细胞生长、成骨分化和体外炎症反应的影响。
Impact of FGF1 on human periodontal ligament fibroblast growth, osteogenic differentiation and inflammatory reaction in vitro.
机构信息
Department of Orthodontics, RWTH Aachen University Hospital, Pauwelsstr. 30, 52074, Aachen, Germany.
Department of Orthodontics, Jena University Hospital, Jena, Germany.
出版信息
J Orofac Orthop. 2022 Oct;83(Suppl 1):42-55. doi: 10.1007/s00056-021-00363-6. Epub 2021 Dec 7.
PURPOSE
To investigate in vitro the impact of fibroblast growth factor 1 (FGF1) in comparison to ascorbic acid (AscA) on human periodontal ligament fibroblast (HPdLF) growth, their osteogenic differentiation, and modulation of their inflammatory reaction to mechanical stress.
METHODS
The influence of different concentrations of FGF1 (12.5-200 ng/mL) on growth and proliferation of HPdLF cells was analyzed over 20 days by counting cell numbers and the percentage of Ki67-positive cells. Quantitative expression analysis of genes encoding the osteogenic markers alkaline phosphatase (ALPL), Runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), and osteopontin (OSP), as well as the fibroblast markers vimentin (VIM) and fibroblast-specific protein 1 (FSP1), was performed after 2 and 20 days of cultivation. Metabolic activity was determined by MTT assay. For comparison with AscA, 50 ng/mL FGF1 was used for stimulation for 2 and 20 days. Cell number, percentage of Ki67-positive cells, and expression of osteoblast- and fibroblast-specific genes were examined. Alkaline phosphatase activity was visualized by NBT/BCIP and calcium deposits were stained with alizarin red. Cytokine (IL‑6, IL‑8, COX2/PGE2) expression and secretion were analyzed by qPCR and ELISA in 6 h mechanically compressed HPdLF cultured for 2 days with FGF1 or ascorbic acid.
RESULTS
Higher concentrations of FGF1 promoted cell proliferation upon short-term stimulation, whereas prolonged treatment induced the expression of osteogenic markers even with low concentrations. AscA promotes cell growth more markedly than FGF1 in short-term cultures, whereas FGF1 induced osteogenic cell fate more strongly in long-term culture. Both factors induced an increased inflammatory response of HPdLF to mechanical compression.
CONCLUSION
Our data suggest that FGF1 promotes an osteogenic phenotype of HPdLF and limits inflammatory response to mechanical forces compared to AscA.
目的
研究成纤维细胞生长因子 1（FGF1）与抗坏血酸（AscA）相比对人牙周膜成纤维细胞（HPdLF）生长、成骨分化的影响，并调节其对机械应激的炎症反应。
方法
通过计数细胞数量和 Ki67 阳性细胞的百分比，分析不同浓度 FGF1（12.5-200ng/ml）对 HPdLF 细胞生长和增殖的影响，培养 20 天。培养 2 天和 20 天后，定量分析碱性磷酸酶（ALPL）、Runt 相关转录因子 2（RUNX2）、骨钙素（OCN）和骨桥蛋白（OSP）等成骨标志物以及波形蛋白（VIM）和纤维蛋白特异性蛋白 1（FSP1）的基因表达。通过 MTT 测定法测定代谢活性。为了与 AscA 进行比较，用 50ng/ml 的 FGF1 刺激 2 天和 20 天。检测细胞数量、Ki67 阳性细胞百分比和成骨细胞和成纤维细胞特异性基因的表达。通过 NBT/BCIP 可视化碱性磷酸酶活性，用茜素红染色钙沉积。用 qPCR 和 ELISA 分析 6 小时机械压缩培养的 HPdLF 中细胞因子（IL-6、IL-8、COX2/PGE2）的表达和分泌，用 FGF1 或 AscA 培养 2 天。
结果
较高浓度的 FGF1 在短期刺激下促进细胞增殖，而长期处理即使在低浓度下也能诱导成骨标志物的表达。与短期培养相比，AscA 更显著地促进细胞生长，而 FGF1 则在长期培养中更强烈地诱导成骨细胞命运。两种因子都增加了 HPdLF 对机械压缩的炎症反应。
结论
我们的数据表明，与 AscA 相比，FGF1 促进 HPdLF 的成骨表型，并限制对机械力的炎症反应。
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