Abnormal bone remodelling activity of dental follicle cells from a cleidocranial dysplasia patient
Abnormal bone remodelling activity of dental follicle cells from a cleidocranial dysplasia patient
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锁骨颅骨发育不良患者牙囊细胞骨重塑活性异常
DOI：
10.1111/odi.12900
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发表时间：
2018
期刊：
影响因子：
3.8
通讯作者：
Zheng Shuguo
中科院分区：
文献类型：
--
作者：
Liu Yang;Zhang Xianli;Sun Xiangyu;Wang Xiaozhe;Zhang Chenying;Zheng Shuguo
目的探讨一种新的锁骨颅发育不良（cleidocranial dysplasia，CCD）致病基因RUNX 2 +/m（Dental follicle cells，DFC）在恒牙延迟萌出中的作用。培养DFCsRUNX 2 +/m，提取DNA用于RUNX 2突变筛查。测量细胞增殖、碱性磷酸酶（ALP）活性、茜素红染色和成骨细胞特异性基因表达，以评估DFCsRUNX 2 +/m的成骨作用。将DFC与外周血单个核细胞（PBMC）共培养，然后进行抗酒石酸酸性磷酸酶（TRAP）染色、真实的实时PCR和蛋白质印迹以评估DFC sRUNX 2 +/m的破骨细胞诱导能力。557G>C）。与正常对照相比，该突变不影响DFCsRUNX 2 +/m的增殖，但下调了成骨相关基因的表达，导致ALP活性和矿化降低。共培养结果显示，DFCsRUNX 2 +/m减少了TRAP+多核细胞的形成和破骨细胞生成相关基因的表达。结论DFCsRUNX 2 +/m可能通过RANK/RANKL/OPG信号通路影响牙萌出过程中的骨重建活性，从而导致CCD患者恒牙萌出障碍。
ObjectivesTo explore the role of dental follicle cells (DFCs) with a novel cleidocranial dysplasia (CCD) causative geneRUNX2mutation (DFCsRUNX2+/m) in delayed permanent tooth eruption.Materials and methodsA CCD patient with typical clinical features was involved in this study. DFCsRUNX2+/mwere cultured and DNA was extracted forRUNX2mutation screening. Measurements of cell proliferation, alkaline phosphatase (ALP) activity, alizarin red staining and osteoblast‐specific genes expression were performed to assess osteogenesis of DFCsRUNX2+/m. Co‐culture of DFCs and peripheral blood mononuclear cells (PBMCs), followed tartrate‐resistant acid phosphatase (TRAP) staining, real‐time PCR and western blot were performed to evaluate osteoclast‐inductive capacity of DFCsRUNX2+/m.ResultsA missenseRUNX2mutation (c. 557G>C) was found in DFCsRUNX2+/mfrom the CCD patient. Compared with normal controls, this mutation did not affect the proliferation of DFCsRUNX2+/m, but down‐regulated the expression of osteogenesis‐related genes, leading to a decrease in ALP activity and mineralisation. Co‐culture results showed that DFCsRUNX2+/mreduced the formation of TRAP+multinucleated cells and the expression of osteoclastogenesis‐associated genes. Furthermore, the mutation reduced the ratio of RANKL/OPG in DFCsRUNX2+/m.ConclusionsDFCsRUNX2+/mdisturbs bone remodelling activity during tooth eruption through RANK/RANKL/OPG signalling pathway and may thus be responsible for impaired permanent tooth eruption in CCD patients.