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.
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。