【作者】 王赛楠； 穆锦全； 雷港； 闫明； 俞艳； 汤春波； 王子露； 于金华； 张光东；

【Author】 Wang Sainan~(,1,*) Mu Jinquan~(,2,*) Lei Gang~(,1) Yan Ming~(,1) Yu Yan~(,1) Tang Chunbo~(,1,3) Wang Zilu~(,1) Yu Jinhua~(,1,4,§)Zhang Guangdong~(,1,4,§)(1 Institute of Stomatology,Nanjing Medical University,140 Hanzhong Road,Nanjing,Jiangsu 210029,China.2 Nanjing Maternal and Child Health Care Hospital,123 Tianfei Street Mochou Road,Nanjing,Jiangsu 210004, China.3 Prosthetic Department,School of Stomatology,Nanjing Medical University,136 Hanzhong Road,Nanjing,Jiangsu 210029,China.4 Endodontic Department,School of Stomatology,Nanjing Medical University,136 Hanzhong Road,Nanjing,Jiangsu 210029, China.)

【机构】 南京医科大学口腔医学院； 南京市妇幼保健医院； 南京医科大学附属口腔医院修复科； 南京医科大学附属口腔医院牙体牙髓科；

【摘要】 目的:研究胰岛素样生长因子1(IGF1)对根尖牙乳头干细胞(SCAPs)增殖及牙骨向分化能力的影响。方法:在体外分别用含有或不含有100ng/mL的IGF1的培养液诱导SCAPs。用MTT法和流式细胞仪检测SCAPs的增殖能力、碱性磷酸酶(ALP)试剂盒检测ALP的活性、茜素红染色观察钙结节的生成能力,用实时定量PCR和蛋白质印迹westernblot分别从基因和蛋白水平上检测成骨及成牙相关指标的变化。将经过和没有经过IGF1诱导的SCAPs细胞团块分别植入大鼠肾被膜下,比较其体内成牙成骨能力。结果:IGF1诱导以后,SCAPs的增殖能力、ALP活性以及基质矿化能力均得到增强。此外相对于没有经过IGF1处理的细胞,处理以后的SCAPs中与成骨相关的基因与蛋白的表达量明显升高,如ALP、RUNX2、Osterix以及骨钙蛋白,但是成牙相关的特异性标志物牙本质涎蛋白和牙本质涎磷蛋白的表达量却有所下降。体内移植实验同样表明经过IGF1处理后的SCAPs细胞团在体内主要生成骨样组织,而没有处理过的细胞团主要生成骨样牙本质样结构。结论:IGF1可以促进根尖牙乳头干细胞的增殖能力及骨向分化能力。这一发现将有助于我们进一步了解SCAPs的分化机制。更多还原

【Abstract】 Objective:The present study was to investigate the proliferation and osteo/ odonto-blastic differentiation potential of stem cells from apical papilla(SCAPs)induced by insulin-like growth factor 1(IGF1).Methods:SCAPs was cultured in DMEM medium supplemented with or without 100ng/mL of IGF1 in vitro.Methyltetrazolium(MTT)assay and flow cytometry(FCM)analysis were performed to determine the proliferation activity of SCAPs.Alkaline phosphatase(ALP)activity was evaluated using an ALP assay kit and alizarin red staining was used to detect the calcium deposition.Moreover,osteo/ odonto-blastic genes and proteins were assessed by real time RT-PCR and western blot respectively.The SCAPs pellets with and without IGF1 treatment were carefully transplanted into the renal capsules of adult rat hosts to evaluate the differentiation in vivo.Results:After treated by IGF1,SCAPs presented increased proliferation,enhanced alkaline phosphatase(ALP)activity and increased matrix mineralization ability.Western blot and quantitive RT-PCR analyses demonstrated that the expression of osteoblast-related proteins and genes(alkaline phosphatase,runt-related transcription factor 2,osterix,and osteocalcin)were significantly up-regulated in IGF1-treated SCAPs, whereas the expression of specific odontoblast markers(dentin sialoprotein and dentin sialophosphoprotein)was down-regulated after 14-day treatment of IGF1.Transplantation results revealed that IGF1-treated SCAPs pellets in vivo gave birth to bone-like tissues while untreated SCAPs pellets mainly generated osteodentin-like structures.Conclusions: IGF1 can promote the proliferation and osteoblastic differentiation of SCAPs,but has negative effect on odontoblastic differentiation.The present study provides new insight into the mechanism of SCAPs differentiation.更多还原