中文摘要：

目的观察不同浓度氟化物在不同时间段对二维（2D）和三维（3D）培养的成纤维细胞胰岛素样生成因子-1（IGF-1）表达的影响,并探讨其在氟化物刺激成纤维细胞成骨功能方面的作用。方法在2D和3D培养系统中,将成纤维细胞L929分为对照组（F-浓度为0mg/L）和6个染氟组（F-浓度分别为0.0001、0.001、0.1、1、10和20mg/L）,在不同染氟时间段（2D培养2、4、24、48和72h,3D培养4、24、48、72和96h）,采用ELISA法检测细胞培养上清中IGF-1蛋白的含量;免疫组化法（IHC）检测染氟成纤维细胞中IGF-1的表达;RT-PCR法检测染氟成纤维细胞IGF-1基因mRNA的转录水平。结果染氟可明显刺激成纤维细胞培养上清中IGF-1蛋白的表达;各染氟组成纤维细胞胞浆IGF-1的阳性着色较对照组均明显增强;氟作用48h,除F-浓度为20mg/L组外,其余各染氟组成纤维细胞IGF-1基因mRNA的转录水平较对照组均有不同程度的增强,但差异均无统计学意义。结论在2D和3D两种培养条件下,氟化物均可明显刺激成纤维细胞IGF-1的表达,提示在氟化物诱导成纤维细胞成骨功能增强的过程中,IGF-1可能发挥着重要作用。

英文摘要：

Objective To observe the effect of treatment with fluoride at various concentrations for various hours on expressions of insulin-like growth factor-1（IGF-1）in two-dimensional（2D）and three-dimensional（3D）cultures,and investigate its role in stimulation of osteoplastic function of fibroblasts（FBs）with fluoride.Methods The FBs in 2D and 3D culture systems were divided into one control and six test groups separately.The FBs in six test groups were added with DMEM containing 0.000 1,0.001,0.1,1,10 and 20 mg /L of fluorine ion respectively,while those in control groups with DMEM containing no fluorine ion.The IGF-1 protein contents in culture supernatants of FBs in 2D culture system were determined 2,4,24,48 and 72 h,while those in 3D culture system,4,24,48,72 and 96 h after treatment with fluoride by ELISA.Meanwhile,the expressions of IGF-1 in FBs treated with fluoride were determined by immunohistochemical（IHC）method,while the transcription of IGF-1 mRNA by RT-PCR.Results The treatment with fluoride stimulated the expression of IGF-1 protein in culture supernatant of FBs significantly.IHC method showed that the color of IGF-1-positive FBs in test groups were significantly deeper than those in control groups.The transcription levels of IGF-1 mRNA in FBs 48 h after treatment with fluoride except that at a concentration of 20 mg /L increased at various degrees,while showed no significant difference with those in control groups.Conclusion Fluoride stimulated the expression of IGF-1 in FBs in both 2D and 3D cultures significantly,indicating that IGF-1 might play an important role in enhancing the osteoplastic function of FBs with fluoride.