骨调节蛋白在人牙髓干细胞成骨/成牙分化中的作用。
The role of osteomodulin on osteo/odontogenic differentiation in human dental pulp stem cells.
机构信息
Department of Endodontics, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
National Clinical Research Center for Oral Diseases, Shanghai, China.
出版信息
BMC Oral Health. 2019 Jan 22;19(1):22. doi: 10.1186/s12903-018-0680-6.
BACKGROUND
Extracellular matrix secretion and odontoblastic differentiation in human dental pulp stem cells (hDPSCs) are the cellular bases for reparative dentinogenesis. Osteomodulin (OMD) is a member of the small leucine-rich proteoglycan family distributed in the extracellular matrix but little is known about its role in osteo/odontogenic differentiation. The objective of this study was to investigate the role of OMD during osteo/odontoblastic differentiation of hDPSCs.
METHODS
hDPSCs were selected using immune-magnetic beads and their capability of multi-differentiation was identified. OMD knockdown was achieved using short hairpin RNA (shRNA) lentivirus and was confirmed by western blot. Gene expression was measured by real-time qPCR and osteo/odontoblastic differentiation of hDPSCs was determined by alizarin red S staining.
RESULTS
Compared with uninduced cells, the transcription of OMD was up-regulated by 35-fold at the late stage of osteo/odontogenic differentiation. shRNA-mediated gene silencing of OMD decreased the expression of odontoblastic genes, such as alkaline phosphatase (ALP), dentin matrix acidic phosphoprotein 1 (DMP1) and dentin sialophosphoprotein (DSPP). Besides, knockdown of OMD attenuated the mineralized nodules formation induced by osteo/odontogenic medium.
CONCLUSIONS
These results implied that OMD may play a pivotal role in modulating the osteo/odontoblastic differentiation of hDPSCs.
背景
人牙髓干细胞（hDPSCs）的细胞外基质分泌和成牙本质细胞分化是修复性牙本质形成的细胞基础。骨调节素（OMD）是小富含亮氨酸的蛋白聚糖家族的成员，分布在细胞外基质中，但对其在成骨/成牙本质分化中的作用知之甚少。本研究旨在探讨 OMD 在 hDPSCs 成骨/成牙本质分化过程中的作用。
方法
使用免疫磁珠分离 hDPSCs，并鉴定其多向分化能力。通过短发夹 RNA（shRNA）慢病毒实现 OMD 敲低，并通过 Western blot 进行验证。通过实时 qPCR 测量基因表达，并通过茜素红 S 染色检测 hDPSCs 的成骨/成牙本质分化。
结果
与未诱导的细胞相比，在成骨/成牙本质分化的晚期，OMD 的转录水平上调了 35 倍。shRNA 介导的 OMD 基因沉默降低了成牙本质基因的表达，如碱性磷酸酶（ALP）、牙本质基质酸性磷酸蛋白 1（DMP1）和牙本质涎磷蛋白（DSPP）。此外，敲低 OMD 减弱了成骨/成牙本质培养基诱导的矿化结节形成。
结论
这些结果表明，OMD 可能在调节 hDPSCs 的成骨/成牙本质分化中发挥关键作用。
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