Arany Szilvia, Koyota Souichi, Sugiyama Toshihiro
Department of Biochemistry, Akita University School of Medicine, Akita, Japan.
J Cell Biochem. 2009 Mar 1;106(4):539-45. doi: 10.1002/jcb.22006.
Contemporary strategies in tooth repair markedly rely on the newest findings on the cellular and biological components of dental development. Among several identified bioactive molecules, neurotrophins were recently proposed to affect tooth germ cell proliferation, differentiation, and cell-extracellular matrix interactions. The present study attempted to explore the effect of nerve growth factor (NGF) on a spontaneously immortalized dental papilla mesenchymal cell line. NGF induced differentiation of odontoblast-lineage cells with subsequent biomineralization in vitro. Here we showed that normalized transcript levels of tissue-specific markers such as DSPP and DMP1 were elevated significantly, indicating cell differentiation and maturation processes. We performed innovative gene expression analysis of TM14, a matricellular protein and novel member of the fibulin family. TM14 expression followed a pattern similar to that of DMP1, which suggests its important role in cell-matrix and intercellular interactions during dentin calcification. Alkaline phosphatase enzyme assay confirmed the extracellular matrix calcifications in NGF-supplemented groups. Thus, NGF was characterized as a potent promoter of mineralization during dentin formation. For the first time, we included TM14 in odontoblast genotype analysis and proved that NGF also promotes in vitro odontoblast differentiation. Collectively, these results highlight the importance of NGF during tooth morphogenesis, as well as urge the elaboration of complex epithelial-mesenchymal tissue cultures, where further elucidation of the signaling factor network could be completed.
当代牙齿修复策略显著依赖于牙齿发育的细胞和生物学成分的最新研究成果。在几种已确定的生物活性分子中,神经营养因子最近被认为会影响牙胚细胞的增殖、分化以及细胞与细胞外基质的相互作用。本研究试图探讨神经生长因子(NGF)对一种自发永生化的牙乳头间充质细胞系的影响。NGF在体外诱导成牙本质细胞系细胞分化并随后发生生物矿化。在此我们表明,诸如DSPP和DMP1等组织特异性标志物的转录水平显著升高,表明细胞分化和成熟过程。我们对TM14进行了创新性的基因表达分析,TM14是一种基质细胞蛋白,也是纤连蛋白家族的新成员。TM14的表达模式与DMP1相似,这表明其在牙本质钙化过程中的细胞-基质和细胞间相互作用中具有重要作用。碱性磷酸酶活性测定证实了补充NGF组的细胞外基质钙化。因此,NGF被确定为牙本质形成过程中矿化的有效促进剂。我们首次将TM14纳入成牙本质细胞基因型分析,并证明NGF也能促进体外成牙本质细胞分化。总的来说,这些结果突出了NGF在牙齿形态发生过程中的重要性,同时也促使人们开展复杂的上皮-间充质组织培养研究,以便进一步阐明信号因子网络。