Oral Health Science Center HRC7, Tokyo Dental College, Chiba, Tokyo, Japan.
Int Endod J. 2013 Jan;46(1):30-9. doi: 10.1111/j.1365-2591.2012.02089.x. Epub 2012 Jun 30.
To investigate the effects of tenascin-C (TN-C) on cultured rat dental pulp cells in relation to the expression of Notch signalling.
Subcultured dental pulp cells derived from rat incisors were seeded both in wells and on plastic coverslips coated with various concentrations of recombinant human TN-C. Expression of bone-related mRNA was then analysed by RT-PCR and observed by immunohistochemical staining. Encoding of Notch1 and Notch2 (markers of initial differentiation of odontoblast-like cells), alkaline phosphatase (ALP), osteopontin (OPN) and osteocalcin (OCN) (markers of mineralization) was investigated. Non-TN-C-coated wells were used as controls. Primary antibodies to Notch1, ALP and OCN were used for immunofluorescence staining, and ALP activity was evaluated. Data were compared using Student's t-test.
Cell proliferation rate in the experimental groups was significantly higher (P < 0.05) than that in the control group at 72 h. Expression of Notch1, Notch2, ALP, OPN and OCN mRNAs was significantly higher (P < 0.05) in the experimental group than that in the control group. Strongly positive staining for Notch1, ALP and OCN was observed in the experimental group. ALP activity was significantly higher (P < 0.01) in the experimental group than in the control group at 24 h.
TN-C promoted differentiation of rat dental pulp cells by the activation of Notch.
研究 tenascin-C（TN-C）对培养的大鼠牙髓细胞的影响，探讨其与 Notch 信号通路表达的关系。
从小鼠切牙分离培养牙髓细胞，分别接种于铺有不同浓度重组人 TN-C 的孔板和塑料盖玻片上。采用 RT-PCR 分析骨相关 mRNA 的表达，并进行免疫组织化学染色观察。检测 Notch1 和 Notch2（成牙本质细胞样细胞初始分化的标志物）、碱性磷酸酶（ALP）、骨桥蛋白（OPN）和骨钙素（OCN）（矿化标志物）的编码。非 TN-C 包被的孔板作为对照。使用 Notch1、ALP 和 OCN 的一抗进行免疫荧光染色，并评估 ALP 活性。采用 Student's t 检验比较数据。
实验组细胞增殖率在 72 h 时明显高于对照组（P<0.05）。实验组 Notch1、Notch2、ALP、OPN 和 OCN mRNA 的表达明显高于对照组（P<0.05）。实验组 Notch1、ALP 和 OCN 染色呈强阳性。实验组在 24 h 时的 ALP 活性明显高于对照组（P<0.01）。
TN-C 通过激活 Notch 促进大鼠牙髓细胞的分化。
