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人乳牙脱落干细胞与牙髓干细胞在体外成骨分化及破骨细胞能力方面的差异

[Difference of in vitro osteogenic differentiation and osteoclast capacity between stem cells from human exfoliated deciduous teeth and dental pulp stem cells].

作者信息

Lu Bo-Wen, Liu Na, Xu Lu-Lu, Shi Hai-Gang, Zhang Yang, Zhang Wei

机构信息

Department of Orthodontics, 2Institute of Oral Medicine, General Hospital of PLA, Beijing 100853, China. E-mail:

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2016 Feb;36(2):180-5.

Abstract

OBJECTIVE

To compare the osteogenic differentiation potential and osteoclast capacity between stem cells from human exfoliated deciduous teeth (SHED) in the physiological root resorption period and dental pulp stem cells (DPSCs).

METHODS

SHED and DPSCs were isolated, purified and cultured in vitro. The two stem cells were examined with ALP staining at 14 days and with alizarin red staining at 21 days of osteogenic induction, and the expressions of the genes associated with osteogenesis and osteoclastogenesis were detected using real-time PCR.

RESULTS

The isolated SHED and DPSCs both showed an elongate spindle-shaped morphology. After osteogenic induction of the cells, Alizarin red staining visualized a greater number of mineralized nodules in SHED than in DPSCs (P<0.05), and SHED also exhibited a stronger ALP activity than DPSCs (P<0.05). RT-PCR test results showed that the two stem cells expressed RANKL,OCN, ALP, OPG and Runx2 mRNA after osteogenic induction, but the expression levels of Runx2, OCN and ALP were lower in DPSCs than in SHED (P<0.05), and the ratio of RANKL/OPG was significantly higher in SHED (P<0.05).

CONCLUSIONS

Compared with DPSCs, SHED has not only the ability of osteogenic differentiation but also an osteoclast capacity, which sheds light on the regulatory role of SHED in physiological root resorption bone remodeling.

摘要

目的

比较生理性牙根吸收期人乳牙脱落干细胞(SHED)和成牙本质细胞干细胞(DPSCs)的成骨分化潜能和破骨细胞能力。

方法

体外分离、纯化和培养SHED和DPSCs。在成骨诱导14天时对两种干细胞进行碱性磷酸酶(ALP)染色,21天时进行茜素红染色,并使用实时定量聚合酶链反应(real-time PCR)检测与成骨和破骨相关基因的表达。

结果

分离出的SHED和DPSCs均呈细长梭形形态。细胞成骨诱导后,茜素红染色显示SHED中矿化结节数量多于DPSCs(P<0.05),且SHED的ALP活性也强于DPSCs(P<0.05)。逆转录-聚合酶链反应(RT-PCR)检测结果显示,两种干细胞在成骨诱导后均表达核因子κB受体活化因子配体(RANKL)、骨钙素(OCN)、碱性磷酸酶(ALP)、骨保护素(OPG)和 Runt相关转录因子2(Runx2)mRNA,但DPSCs中Runx2、OCN和ALP的表达水平低于SHED(P<0.05),且SHED中RANKL/OPG的比值显著更高(P<0.05)。

结论

与DPSCs相比,SHED不仅具有成骨分化能力,还具有破骨细胞能力,这为SHED在生理性牙根吸收骨重塑中的调节作用提供了线索。

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