人牙髓干细胞在钛金属表面向成骨细胞样细胞分化的实验研究  

Differentiation of osteoblast-like cells from human dental pulp stem cells on titanium-alloy

作　　者：张晓艳[1] 李小彤[2] 曾祥龙[2] 

机构地区：[1]北京首都医科大学口腔医学院正畸科,100050 [2]北京大学口腔医学院正畸科

出　　处：《北京口腔医学》2015年第5期249-253,共5页Beijing Journal of Stomatology

摘　　要：目的探讨矿化液诱导条件下人牙髓干细胞(human dental pulp stem cells,HDPSCs)在钛金属表面向成骨细胞样细胞分化的潜力。方法将HDPSCs接种于钛金属板表面,于第3、7d时观察其增殖状态。将HDPSCs接种于钛金属板表面进行矿化液诱导,于诱导的第0、3、5、7、14、21、28d时,采用RT-PCR检测其牙本质涎磷蛋白(DSPP)、碱性磷酸酶(ALP)、骨涎蛋白(BSP)和骨钙素(OCN)的基因表达。Western印迹分析、免疫荧光检测BSP及OCN蛋白表达。对照组为矿化液诱导培养皿中的HDPSCs,检测指标、方法基本相同,免疫细胞化学染色检测OCN蛋白表达。结果钛金属板上HDPSCs增殖迅速,表现出良好的生物相容性。两组细胞均自诱导第3天ALPmRNA呈高表达、DSPP始终不表达。培养皿组BSP、OCNmRNA于第5天开始表达,并随时间延长而表达增高;钛金属组BSP、OCNmRNA表达于第14天出现第一峰值,随后有所下降,28天再次升高。两组BSP蛋白表达趋势与其RT-PCR结果一致,第5天OCN染色阳性。结论钛金属表面HDPSCs经矿化液诱导向成骨细胞样细胞分化。Objective To induce human dental pulp stem cells （HDPSCs） on titanium-alloy into osteoblast-like cells by mineralizing culture medium. Methods HDPSCs were seeded on titanium-alloy surfaces, the proliferation of the cells was compared on the 34 and 7th day respectively. HDPSCs on titanium-alloy were cultured by mineralizing culture medium for 28 days. The gene expression of ALP, DSPP, BSP and OCN on day 0,3,5,7,14,21 and 28 was evaluated by RT-PCR. The protein expression of BSP and OCN was analyzed by Western blotting and immunofluorescence. The HDPSCs cultured by the same medium in normal dish served as control. The gene and protein expression for the same osteoblast-like markers was examined by RT-PCR and OCN was examined with immunocytochemistry. Results HDPSCs attached to titanium-alloy and proliferated well. ALP mRNA in both groups was detected since the 3rd day, but DSPP mRNA was not expressed. The effects of titanium and culturing dish on differentiation of HDPSCs. The gene of BSP and OCN in dish group was expressed since the 52 day and exhibited increment with time. HDPSCs on titanium-alloy also expressed the gene of BSP and OCN since the 52 day, and the expression achieved the first peak by the 14th day, then decreased to some extent but increased again by the 28th day. The expression trend of BSP protein was consistent with BSP mRNA. OCN was stained positively since the 5th day in two groups. Conclusion HDPSCs on titanium-alloy could be induced to differentiate into osteoblast-like cells, which suggests that HDPSCs on titanium-alloy may be a potential source of cells used for strengthening the implants.

关 键 词：人牙髓干细胞 钛金属 成骨细胞样细胞 

分 类 号：R781[医药卫生—口腔医学]