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目的:研究已证实,碱性成纤维细胞生长因子刺激牙周膜细胞后可促进人牙周膜细胞的增殖,以利于重建丧失的牙周组织。利用不同质量浓度碱性成纤维细胞生长因子刺激体外培养的人正常牙周膜细胞,观察牙周膜细胞内核心蛋白多糖基因表达。方法:采用胰酶消化法分离培养牙周膜细胞。取第3代对数生长期的细胞,加入含有10%二甲基亚砜和含体积分数20%胎牛血清冻存液的DMEM中进行冻存,第2天移入液氮中保存。免疫组织化学方法行抗波形丝蛋白、角蛋白染色鉴定。取第6代人牙周膜细胞,分为实验组和空白组。实验组分别用含质量浓度为0.1,1,10μg/L碱性成纤维细胞生长因子的DMEM培养液标准条件下培养24h;空白组用DMEM培养液标准条件下培养24h。RT-PCR法检测细胞内核心蛋白多糖基因表达变化。结果:镜下观察空白组细胞未见明显变化,实验组可见细胞增殖,刺激前瓶底细胞较稀疏的区域变得密集了。加入碱性成纤维细胞生长因子的牙周膜细胞内核心蛋白多糖的mRNA表达水平明显下降了,而且随着碱性成纤维细胞生长因子质量浓度的增加,抑制作用逐渐减弱,在0.1μg/L时抑制作用最强,在10μg/L时抑制作用最弱。结论:碱性成纤维细胞生长因子刺激可促进牙周膜细胞增殖,呈剂量依赖性抑制核心蛋白多糖的合成,0.1μg/L时抑制作用最强。
OBJECTIVE: It has been confirmed that the proliferation of human periodontal ligament cells can be promoted by basic fibroblast growth factor stimulating periodontal ligament cells in order to reconstruct lost periodontal tissues. Human normal human periodontal ligament cells cultured in vitro were stimulated with different concentrations of basic fibroblast growth factor to observe the expression of decorin in periodontal ligament cells. Methods: The periodontal ligament cells were isolated and cultured by trypsin digestion. The third generation of logarithmic growth phase cells were added into DMEM containing 10% dimethylsulfoxide and 20% fetal bovine serum in cryogenic solution, and stored in liquid nitrogen on the second day. Immunohistochemical staining of silk fibroin, keratin staining. Sixth generation human periodontal ligament cells were divided into experimental group and blank group. The experimental group were incubated with DMEM containing 0.1, 1, 10μg / L basic fibroblast growth factor for 24 hours under standard conditions. The blank group was cultured for 24 hours under the standard conditions with DMEM culture medium. RT-PCR method was used to detect the changes of intracellular decorin gene expression. Results: No significant changes were observed in the blank group under the microscope. The proliferation of cells in the experimental group was observed. The expression of decorin mRNA in periodontal ligament cells with basic fibroblast growth factor decreased significantly, and with the increase of basic fibroblast growth factor concentration, the inhibitory effect gradually decreased. At the concentration of 0.1μg / L When the inhibitory effect of strongest, at 10μg / L, the weakest inhibition. Conclusion: BfF stimulation can promote the proliferation of periodontal ligament cells and inhibit the synthesis of decorin in a dose - dependent manner. The inhibitory effect of 0.1μg / L is the strongest.