PDF(Pubmed)
Abstract：
Introduction: Root dentin formation is an important process in tooth development. We tried to identify potential genes that regulate root dentin formation which could be potentially used for the regeneration and repair of defective or damaged dental roots. Methods: Tissues harvested from the labial and lingual sides of mouse incisors were used for microarray analysis. Gene ontology (GO) analysis of differentially expressed genes indicated the critical role of extracellular matrix in the discrepancy of dentin formation between root and crown, for which hemicentin-1 (Hmcn1) was selected as the target gene. Single-cell RNA sequencing analysis the expression pattern of Hmcn1 at different developmental stages in mouse molars. The spatiotemporal expression of HMCN1 in mouse incisors and molars was detected by immunohistochemical staining. The functions of HMCN1 in human dental pulp cells, including proliferation, differentiation and migration, were examined in vitro by CCK8 assay, BrdU assay, wound-healing assay, ALP staining and alizarin red staining, respectively. Results: It was showed that HMCN1 expression was more pronounced in papilla-pulp on the root than crown side in mouse incisors and molars. In vitro experiments presented inhibited dentinogenesis and migration after HMCN1-knockdown in human dental pulp cells, while there was no significant difference in proliferation between the HMCN1-knockdown group and control group. Discussion: These results indicated that HMCN1 plays an important role in dentinogenesis and migration of pulp cells, contributing to root dentin formation.
摘要：
简介：牙根牙本质形成是牙齿发育的重要过程。我们试图确定调节牙根牙本质形成的潜在基因，这些基因可潜在地用于有缺陷或受损牙根的再生和修复。方法：从小鼠切牙的唇侧和舌侧收获的组织用于微阵列分析。差异表达基因的基因本体论（GO）分析表明，细胞外基质在牙根和牙冠之间牙本质形成差异中的关键作用。选择半形蛋白-1（Hmcn1）作为靶基因。单细胞RNA测序分析Hmcn1在小鼠磨牙不同发育阶段的表达模式。免疫组化染色检测HMCN1在小鼠切牙和磨牙中的时空表达。HMCN1在人牙髓细胞中的功能,包括扩散，分化和迁移，通过CCK8测定法进行体外检查，BrdU测定,伤口愈合试验，ALP染色和茜素红染色，分别。结果：结果表明，HMCN1在小鼠切牙和磨牙的根部乳头牙髓中的表达比冠侧更明显。体外实验表明，在人牙髓细胞中HMCN1敲低后，抑制牙本质生成和迁移，而HMCN1敲除组与对照组之间的增殖没有显着差异。讨论：这些结果表明，HMCN1在牙本质形成和牙髓细胞迁移中起重要作用，有助于牙根牙本质的形成。
