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Abstract：
BACKGROUND: The aim of this study was to investigate the role of the steroid fluocinolone acetonide on the proliferation and mineralization of human dental pulp cells (DPCs). The potential effect of fluocinolone acetonide on reparative dentin formation and the recovery of injured dental pulp were evaluated.
METHODS: The proliferative effect of fluocinolone acetonide on DPCs was analyzed by cholecystokinin octapeptide assay and flow cytometry. The mineralized effect of fluocinolone acetonide was investigated by the detection of mineralization-related biomarkers including alkaline phosphatase (ALP), bone sialoprotein, and osteocalcin by using ALP histochemical staining, ALP activity, immunostaining, alizarin red staining, and reverse-transcriptase polymerase chain reaction. The molecules, including dentin sialophosphoprotein and Wnt4, involved in the process of mineralization were detected by real-time polymerase chain reaction and Western blot analysis.
RESULTS: Low concentrations of fluocinolone acetonide (0.1-40 μmol/L) promoted the proliferation of DPCs. The flow cytometry results showed that the CD146-positive subpopulation of DPCs was significantly increased after treatment with fluocinolone acetonide at 1 and 10 μmol/L for 48 hours, respectively. The messenger RNA expression and activity of the early-stage mineralization marker ALP were evidently increased in fluocinolone acetonide-treated DPCs compared with the untreated control group, so did the middle-stage mineralization marker bone sialoprotein and the late-stage mineralization marker osteocalcin. Meanwhile, Wnt4 and the dentin-specific marker dentin sialophosphoprotein were obviously up-regulated by fluocinolone acetonide compared with the untreated controls.
CONCLUSIONS: Fluocinolone acetonide can promote the proliferation of DPCs, especially for the CD146+ subpopulation. Fluocinolone acetonide can initiate the mineralization of DPCs and has the potential role in repairing injured pulp tissues.
METHODS: The proliferative effect of fluocinolone acetonide on DPCs was analyzed by cholecystokinin octapeptide assay and flow cytometry. The mineralized effect of fluocinolone acetonide was investigated by the detection of mineralization-related biomarkers including alkaline phosphatase (ALP), bone sialoprotein, and osteocalcin by using ALP histochemical staining, ALP activity, immunostaining, alizarin red staining, and reverse-transcriptase polymerase chain reaction. The molecules, including dentin sialophosphoprotein and Wnt4, involved in the process of mineralization were detected by real-time polymerase chain reaction and Western blot analysis.
RESULTS: Low concentrations of fluocinolone acetonide (0.1-40 μmol/L) promoted the proliferation of DPCs. The flow cytometry results showed that the CD146-positive subpopulation of DPCs was significantly increased after treatment with fluocinolone acetonide at 1 and 10 μmol/L for 48 hours, respectively. The messenger RNA expression and activity of the early-stage mineralization marker ALP were evidently increased in fluocinolone acetonide-treated DPCs compared with the untreated control group, so did the middle-stage mineralization marker bone sialoprotein and the late-stage mineralization marker osteocalcin. Meanwhile, Wnt4 and the dentin-specific marker dentin sialophosphoprotein were obviously up-regulated by fluocinolone acetonide compared with the untreated controls.
CONCLUSIONS: Fluocinolone acetonide can promote the proliferation of DPCs, especially for the CD146+ subpopulation. Fluocinolone acetonide can initiate the mineralization of DPCs and has the potential role in repairing injured pulp tissues.
摘要：
背景：这项研究的目的是研究类固醇氟轻松酮对人牙髓细胞（DPC）增殖和矿化的作用。评价了氟轻松对修复性牙本质形成和损伤牙髓恢复的潜在影响。
方法：采用八肽胆囊收缩素法和流式细胞术分析氟轻松对DPCs的增殖作用。通过检测与矿化相关的生物标志物，包括碱性磷酸酶（ALP），研究了氟轻松的矿化作用。骨唾液蛋白，和骨钙蛋白通过使用ALP组织化学染色，ALP活性,免疫染色，茜素红染色，和逆转录酶聚合酶链反应。分子，包括牙本质唾液酸磷蛋白和Wnt4，参与矿化过程，通过实时聚合酶链反应和Westernblot分析检测。
结果：低浓度的氟轻松（0.1-40μmol/L）促进了DPCs的增殖。流式细胞术结果显示,1和10μmol/L氟轻松处理48小时后,DPCs的CD146阳性亚群明显增多,分别。与未处理的对照组相比，氟轻松处理的DPC中早期矿化标记ALP的信使RNA表达和活性明显增加。中期矿化标记骨唾液酸蛋白和晚期矿化标记骨钙蛋白也是如此。同时,与未处理的对照相比，氟轻松明显上调了Wnt4和牙本质特异性标记牙本质唾液酸磷蛋白。
结论：氟轻松可以促进DPCs的增殖，特别是对于CD146+亚群。氟轻松可以启动DPC的矿化，并在修复损伤的牙髓组织中具有潜在的作用。
方法：采用八肽胆囊收缩素法和流式细胞术分析氟轻松对DPCs的增殖作用。通过检测与矿化相关的生物标志物，包括碱性磷酸酶（ALP），研究了氟轻松的矿化作用。骨唾液蛋白，和骨钙蛋白通过使用ALP组织化学染色，ALP活性,免疫染色，茜素红染色，和逆转录酶聚合酶链反应。分子，包括牙本质唾液酸磷蛋白和Wnt4，参与矿化过程，通过实时聚合酶链反应和Westernblot分析检测。
结果：低浓度的氟轻松（0.1-40μmol/L）促进了DPCs的增殖。流式细胞术结果显示,1和10μmol/L氟轻松处理48小时后,DPCs的CD146阳性亚群明显增多,分别。与未处理的对照组相比，氟轻松处理的DPC中早期矿化标记ALP的信使RNA表达和活性明显增加。中期矿化标记骨唾液酸蛋白和晚期矿化标记骨钙蛋白也是如此。同时,与未处理的对照相比，氟轻松明显上调了Wnt4和牙本质特异性标记牙本质唾液酸磷蛋白。
结论：氟轻松可以促进DPCs的增殖，特别是对于CD146+亚群。氟轻松可以启动DPC的矿化，并在修复损伤的牙髓组织中具有潜在的作用。
