The role of dentin matrix protein 1 (DMP1) in regulation of osteogenic differentiation of rat dental follicle stem cells (DFSCs).
The role of dentin matrix protein 1 (DMP1) in regulation of osteogenic differentiation of rat dental follicle stem cells (DFSCs).
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DOI：
10.1016/j.archoralbio.2014.12.013
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发表时间：
2015-04
影响因子：
3
通讯作者：
Yao, Shaomian
中科院分区：
文献类型：
--
作者：
Rad, Maryam Rezai;Liu, Dawen;He, Hongzhi;Brooks, Hunter;Xiao, Mei;Wise, Gary E.;Yao, Shaomian
关键词：
原代分离的牙囊干细胞(DFSCs)具有很强的成骨能力，但在体外培养过程中这种能力会减弱。由于牙本质基质蛋白1(DMP1)在成骨细胞的成熟过程中起着至关重要的作用，因此我们的目标是：(1)DMP1在DFSCs中的表达；(2)DMP1表达与DFSCs成骨能力的相关性；(3)DMP1促进DFSCs成骨分化的能力。DFSCs及其非干细胞对应的牙囊细胞(DFC)取自出生后的大鼠幼鼠。用实时荧光定量RT-PCR和Western blotting检测DFSCs和DFC中DMP1的表达。对不同代次的DFSCs进行成骨诱导。分析成骨与DMP1表达的相关性。然后，用siRNA下调DFSCs中DMP1的表达，然后进行成骨诱导，以评价DMP1基因敲除的效果。最后，在含有小鼠重组DMP1(MrDMP1)的成骨诱导液中培养DMP1表达降低和成骨能力降低的晚期DFSCs，以确定DMP1是否能恢复DFSCs的成骨。DFSCs的DMP1表达水平明显高于DFC。DMP1的表达与DFSCs的成骨能力相关。抑制DMP1的表达明显降低DFSCs的成骨能力和成骨基因的表达，而在成骨诱导培养液中加入mrDMP1蛋白则促进成骨。DMP1在DFSCs中高表达，在非干细胞DFC中低表达。DMP1在DFSCs的成骨分化过程中起重要作用。
Primary isolated dental follicle stem cells (DFSCs) possess a strong osteogenesis capability, and such capability is reduced during in vitro culture. Because dentin matrix protein 1 (DMP1) is essential in the maturation of osteoblasts, our objectives were to determine (1) the expression of DMP1 in the DFSCs, (2) the correlation between DMP1 expression and osteogenic capability of DFSCs, and (3) the ability of DMP1 to promote osteogenic differentiation of DFSCs. DFSCs and their non-stem cell counterpart dental follicle cells (DFC) were established from postnatal rat pups. Expression of DMP1 in the DFSCs and DFC was determined using real-time RT-PCR and western blotting. Different passages of DFSCs were subjected to osteogenic induction. The correlation between osteogenesis and DMP1 expression was analyzed. Then, expression of DMP1 in the DFSCs was knocked-down using siRNA, followed by osteogenic induction to evaluate the effect of DMP1-knockdown. Finally, the late passage DFSCs with reduced DMP1 expression and osteogenic capability were cultured in osteogenic induction medium containing mouse recombinant DMP1 (mrDMP1) to determine if DMP1 can restore osteogenesis of DFSCs. DFSCs expressed much higher levels of DMP1 than did DFC. DMP1 expression was correlated with the osteogenic capability of DFSCs. Knockdown of DMP1 expression markedly decreased the osteogenesis and osteogenic gene expression in the DFSCs whereas adding mrDMP1 protein to the osteogenic induction medium enhanced osteogenesis. DMP1 is highly expressed in the DFSCs, but minimally expressed in non-stem cell DFC. DMP1 appears to play an important role for osteogenic differentiation of the DFSCs.
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