Pan Gang, Zhou Qianwen, Pan Chenhua, Zhang Yingxue
Department of Stomatology, PuRen Hospital, Wuhan University of Science and Technology, Wuhan, 430081, Hubei Province, China.
Biological Cell Therapy Research Center, PuRen Hospital, Wuhan University of Science and Technology, Wuhan, 430081, Hubei Province, China.
Cell Biochem Biophys. 2025 Mar;83(1):507-517. doi: 10.1007/s12013-024-01480-w. Epub 2024 Aug 16.
Dental pulp stem cells (DPSCs) are a class of cells with the potential of self-replication and multi-directional differentiation, which are widely considered to have great application value. It was to investigate miR-586 in DPSCs differentiated into odontoblast-like cells. In this article, human dental pulp stem cells (hDPSCs) were used as samples, and hDPSCs were co-cultured with endothelial progenitor cells (EPCs). Furthermore, a lentiviral expression vector for the miR-586 inhibitor was established. The effect of miR-586 inhibitor expression vector on the activity of hDPSCs was detected by Cell Counting Kit-8 (CCK-8). The differentiation of hDPSCs was tested by mineralized nodule staining. The expression of miR-586 and a gene related to dental cell differentiation in the pulp was subjected to detection by real-time quantitative PCR (qRT-PCR). As against the normal hDPSCs and the empty vector, the miR-586 lentivirus expression inhibition vector could visibly raise the expression of dentin sialophosphoprotein (DSPP) in hDPSCs; and the cell proliferation activity was visibly enhanced; In addition, the mRNA expressions of dentin-matrix acidic phosphoprotein 1 (DMP-1) and alkaline phosphatase (ALP) were visibly raised in the miR-586 lentivirus expression inhibition vector (all P < 0.05). Additionally, ALP activity was significantly enhanced (P < 0.05). The number of mineralized nodules was significantly increased (P < 0.05). MiR-586 plays a key regulatory function in DPSCs differentiated into odontoblast-like cells and is associated with specific molecular mechanisms.
牙髓干细胞(DPSCs)是一类具有自我复制和多向分化潜能的细胞,被广泛认为具有巨大的应用价值。本研究旨在探讨miR-586在牙髓干细胞向成牙本质细胞样细胞分化过程中的作用。本文以人牙髓干细胞(hDPSCs)为样本,将其与内皮祖细胞(EPCs)共培养。此外,构建了miR-586抑制剂的慢病毒表达载体。采用细胞计数试剂盒-8(CCK-8)检测miR-586抑制剂表达载体对hDPSCs活性的影响。通过矿化结节染色检测hDPSCs的分化情况。采用实时定量PCR(qRT-PCR)检测miR-586及牙髓中与牙细胞分化相关基因的表达。与正常hDPSCs和空载体相比,miR-586慢病毒表达抑制载体可显著提高hDPSCs中牙本质涎磷蛋白(DSPP)的表达;细胞增殖活性明显增强;此外,miR-586慢病毒表达抑制载体中牙本质基质酸性磷酸蛋白1(DMP-1)和碱性磷酸酶(ALP)的mRNA表达明显升高(均P < 0.05)。另外,ALP活性显著增强(P < 0.05)。矿化结节数量显著增加(P < 0.05)。MiR-586在牙髓干细胞向成牙本质细胞样细胞分化过程中起关键调节作用,并与特定分子机制相关。