矿化液诱导人前磨牙牙髓干细胞向成骨细胞样细胞的分化  

Induction of human dental pulp stem cells of premolars into osteoblast-like cells by mineralizing culture medium

作　　者：张晓艳[1] 李小彤[1] 曾祥龙[1] 

机构地区：[1]北京大学口腔医学院.口腔医院口腔正畸科,北京100081

出　　处：《上海口腔医学》2010年第4期398-402,共5页Shanghai Journal of Stomatology

基　　金：国家自然科学基金(30772452)~~

摘　　要：目的:探讨矿化液诱导人前磨牙牙髓干细胞(HDPSCs)向成骨细胞样细胞分化的潜力。方法:矿化液诱导前磨牙HDPSCs 28d,于诱导的第7、28天时,应用酶化学染色检测碱性磷酸酶(ALP)表达;第14、21、28天时,应用茜素红染色观察细胞矿化情况;于诱导的第0、3、5、7、14、21、28天时,采用RT-PCR法检测其ALP、牙本质涎磷蛋白(DSPP)、骨涎蛋白(BSP)和骨钙素(OCN)的基因表达;Western印迹分析、免疫细胞化学染色检测BSP及OCN蛋白的表达。结果:经矿化液诱导后,前磨牙HDPSCs ALP染色阳性,形成钙结节。诱导第3天,细胞ALP mRNA呈高表达;DSPP始终不表达;BSP、OCN mRNA于第5天开始表达,并随时间延长而表达增高;BSP蛋白表达趋势与其RT-PCR结果一致;第5天时OCN染色阳性。结论:前磨牙牙髓干细胞具有向成骨细胞样细胞分化的潜能,有望成为骨组织工程的种子细胞。PURPOSE： To induce human dental pulp stem cells（HDPSCs） of premolars into osteoblast-like cells by mineralizing culture medium.METHODS： HDPSCs of premolars were induced by mineralizing culture medium for 28 days.The activity of ALP was examined by enzyme histochemical staining on the 7th and 28th day.The ability of mineralization of HDPSCs was detected by Alizarin-red staining on the 14th,21st and 28th day.The gene expression of ALP,DSPP,BSP,OCN on day 0,3,5,7,14,21 and 28 was evaluated by RT-PCR.The protein expression of BSP,OCN was analyzed by Western blotting and immunocytochemistry.RESULTS： ALP was expressed and mineralized nodules were observed after induction of HDPSCs.ALP mRNA was detected since the 3rd day,while no expression of DSPP mRNA.The gene of BSP,OCN was expressed from the 5th day and exhibited increment with time.The expression trend of BSP protein was consistent with BSP mRNA.OCN stained positive since the 5th day.CONCLUSIONS： HDPSCs of premolars could be induced to differentiate into osteoblast-like cells,which suggests that HDPSCs of premolars may be a potential source of cells used for bone-tissue engineering in the future.

关 键 词：人牙髓干细胞 成骨细胞样细胞 分化 

分 类 号：R780.2[医药卫生—口腔医学]