南方医科大学学报

南方医科大学学报 ›› 2016, Vol. 36 ›› Issue (02): 180-.

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人脱落乳牙牙髓干细胞与人恒牙牙髓干细胞成骨分化及破骨能力的差异

路博闻，刘娜，徐璐璐，石海刚，张洋，张维   

  • 出版日期:2016-02-20 发布日期:2016-02-20

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

  • Online:2016-02-20 Published:2016-02-20

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摘要： 目的比较处于生理性根吸收时期的自然脱落乳牙牙髓干细胞（SHED）和恒牙牙髓干细胞（DPSCs）的成骨分化及破骨能
力，探讨两者在成骨和破骨相关分子的表达情况。方法体外分离、培养和纯化处于生理性根吸收时期的自然脱落乳牙牙髓干
细胞与恒牙牙髓干细胞；两种干细胞在成骨诱导14 d 和21 d，分别进行ALP染色、茜素红染色，并通过Real-time PCR检测
SHED和DPSCs矿化诱导后的成骨与破骨相关基因表达。结果倒置显微镜下观察，SHED和DPSCs形态均为长梭形；两种干
细胞经矿化诱导后，茜素红染色可见SHED的矿化结节计数多于DPSCs（P<0.05），SHED的ALP活性亦强于DPSCs（P<0.05）。
RT-PCR检测结果显示，经矿化诱导后两种干细胞均表达成骨与破骨相关基因Runx2、OCN、ALP、OPG和RANKL，但DPSCs的
Runx2、OCN 和ALP的表达水平均低于SHED（P<0.05），且SHED反应细胞破骨/成骨能力的RANKL/OPG 的比值显著高于
DPSCs（P<0.05）。结论与DPSCs相比，SHED不仅具有较强的成骨分化能力，还兼有较强的破骨能力，为SHED参与生理性根
吸收骨改建的调控机制提供实验依据。

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). Conclusion 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.