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目的:研究人牙囊细胞成骨分化前后LIM矿化蛋白-1的表达。探讨LIM矿化蛋白-1在牙囊细胞向牙周组织分化过程中的作用。方法:组织块加酶消化法分离培养人牙囊细胞,实验组培养液中加入矿化诱导剂,对照组不加矿化诱导剂。培养4周后,免疫细胞化学检测骨涎蛋白的表达;Von Kossa染色检测矿化结节的形成。矿化诱导前后,采用RT-PCR技术检测LIM矿化蛋白-1。结果:培养4周,实验组矿化结节形成,骨涎蛋白免疫细胞化学呈阳性表达,Von Kossa染色阳性;对照组未形成矿化结节,骨涎蛋白表达弱阳性,Von Kossa染色阴性。RT-PCR结果显示LIM矿化蛋白-1在实验组强阳性表达,对照组弱表达。结论:LIM矿化蛋白-1与胚胎期人牙囊细胞的体外矿化过程相关,提示LIM矿化蛋白-1可能在胚胎期牙囊细胞的分化、矿化中发挥一定作用。
Objective: To study the expression of LIM mineralization protein-1 before and after osteogenic differentiation of human dental follicle cells. To investigate the role of LIM mineralized protein-1 in the differentiation of periodontal tissues into periodontal tissues. Methods: Human dental follicle cells were isolated and cultured by enzymatic digestion method. The mineralization inducer was added to the experimental group culture medium and the mineralization inducer was not added in the control group. Four weeks after culture, immunocytochemistry was used to detect the expression of bone sialoprotein. Von Kossa staining was used to detect the formation of mineralized nodules. Before and after mineralization induction, LIM mineralization protein-1 was detected by RT-PCR. Results: After cultured for 4 weeks, the mineralized nodules were formed in the experimental group, the expression of bone sialoprotein immunocytochemistry was positive and Von Kossa staining was positive. The mineralized nodules were not formed in the control group. The expression of bone sialoprotein was weakly positive and Von Kossa staining was negative. RT-PCR results showed that LIM mineralized protein-1 was strongly expressed in the experimental group and weakly in the control group. CONCLUSION: LIM mineralization protein-1 is associated with in vitro mineralization of human dental follicle cells, suggesting that LIM mineralized protein-1 may play a role in the differentiation and mineralization of dental follicle cells in embryonic stage.