BMP-2和地塞米松对体外培养大鼠牙囊细胞矿化能力的影响

目的:探讨BMP-2和地塞米松联合作用对大鼠牙囊细胞分化能力的影响,为牙囊细胞在牙周组织工程中的应用提供实验依据。方法:取生长状态良好的第3代大鼠牙囊细胞,血清饥饿同步化后,分别加入含BMP-2(100ng/ml)、地塞米松Dex(10-8mol/ml)、BMP-2(100ng/ml)+地塞米松(10-8mol/ml)的DMEM培养液,通过碱性磷酸酶(ALP)检测试剂盒、钙化结节染色分别检测不同诱导条件下对大鼠牙囊细胞分化的影响。结果:经BMP-2、Dex、BMP-2+Dex诱导后,体外培养的大鼠牙囊细胞碱性磷酸酶活性均显著高于未诱导组,各组间ALP活性与对照组相比差异有统计学意义(P<0.01)。BMP-2+Dex诱导组ALP活性最强,与BMP-2诱导组间差异有统计学意义(P<0.01),而与Dex诱导组间ALP活性则差异无统计学意义(P>0.01)。培养14d时,BMP-2+Dex诱导组牙囊细胞ALP活性增强最为显著,与BMP-2诱导组及Dex诱导组相比差异均有统计学意义(P<0.01)。矿化结节计数分析显示,各组细胞矿化结节形成的数量及面积明显存在显著的差异,BMP-2、Dex、BMP-2+Dex诱导组与未诱导组相比,其矿化结节形成量差异均有统计学意义(P<0.01),BMP-2+Dex诱导组矿化结节形成量明显大于BMP-2和Dex单独诱导组。结论:BMP-2、地塞米松均能促进牙囊细胞的分化,然而两者联合作用促分化的能力最为显著,提示其在牙周组织工程中的应用前景。

Effect of Bone morphogenetic protein-2 and Dexamethasone on mineralization capability of rat dental follicle cells

Objective： The present study was conducted to investigate the combined effect of bone morphogenetic pro- tein-2 （BMP-2） and dexamethasone （Dex） on the proliferation and differentiation of RDFCs. Methods： The third passage RDFCs were induced by BMP-2 （100ng/ml）, Dex （10^-8mol/ml） and a mixture ofBMP-2 （100ng/ml） and Dex （10hnol/ml） in DMEM, respectively. After 7, 14 days, differentiation of the attached cells were measured with alkaline phosphatase （ALP） activity measurement kit, and mineralization potential was studied by Alizarin Red staining. Results： The results showed that with treatment of BMP-2, Dex alone could promote the differentiation of RDFCs. Greater proliferation of RDFCs was found in combined treatment group at different time points compared with individual treatment groups. Treatment with Dex alone could also augment the expression of ALP. The above results suggested that BMP-2 and Dex could synergistically promote the differentiation of RDFCs into osteoblast. Conclusions： The exposure of Dex as well as BMP-2 to RDFCs with an appropriate concentration promoted osteogenic expression without reverse effects on cell proliferation, which indicated the great potential value in cell-based strategy of bone tissue engineering.