摘要
目的研究骨桥蛋白(osteopontin,OPN)对矿化液诱导牙髓干细胞(dental pulp stem cells,DPSCs)成骨分化能力的影响,并优化其诱导条件。方法矿化液培养DPSCs作为对照组,在矿化液培养基础上添加适宜浓度OPN作为实验组,在倒置相差显微镜下观察诱导后的DPSCs形态变化;应用茜素红染色法检测矿化结节的形成;采用反转录聚合酶链反应(reverse transcription polymerase chain reaction,RT-RCR)分别检测DPSCs骨涎蛋白(BSP)、Runt相关转录因子2(Runx-2)、骨钙蛋白(OCN)、Ⅰ型胶原(Col-1)等骨源性基因表达情况。结果茜素红染色显示诱导培养28 d后两组均出现矿化结节,且实验组的数量和大小明显高于对照组。培养7 d后两组DPSCs骨源性基因表达均成阳性,且实验组中BSP、Runx-2、Col-1及OCN等基因表达水平均明显高于对照组。结论 OPN能增强矿化液诱导DPSCs成骨分化的能力。
Objective To study the influence of osteopontin （OPN） onosteogenetic differentiation ability of dental pulp stem cells （DPSCs） indueed by mineralizing culture medium, and to optimize its inducement conditions. Methods DPSCs cultured in minerali- zing culture mediumworked as control group, and DPSCs cultured in mineralizing culture medium with OPN was the experimental group. The morphological changes of DPSCs were observed by the inverted phase contrast microscope; and the formation of mineralize nodules was detected with Alizarin red staining method; the expression levels of osteogenie related genes including bone sialoprotein （BSP） , Runt-related transcription factor 2 （Runx-2） , osteoealcin （OCN） and collagen-1 （ Col-1 ） were detected by reverse transcrip- tion polymerase chain reaetion （RT-RCR）. Results Alizarin red staining showed that mineralized nodules have formed after being in- duced for 28 days in both groups, and the number and size of the mineralized nodules in experimental group were obviously larger than that in control group. The expressions of osteogenic related genes were all positive in both groups after 7 days. Expression levels of BSP, Runx -2, Col-land OCN genes of experimental group were significantly higher than control group. Conclusions OPN can en- hance the osteogenetic differentiation ability of dental pulp stem ceils （DPSCs） induced by mineralizing culture medium.
出处
《口腔医学》
CAS
2014年第8期566-569,共4页
Stomatology
基金
黑龙江省自然科学基金资助项目(H2013101)
关键词
骨桥蛋白
矿化液
牙髓干细胞
分化
osteopontin
mineralizing culture medium
dental pulp stem cells
differentiation
