人脱落乳牙牙髓干细胞与牙髓干细胞生物学功能差异研究

Research differences of biological functions between stem cells from human exfoliated deciduous teeth and dental pulp stem cells

目的:从不同角度对人乳牙牙髓干细胞(SHED)与恒牙牙髓干细胞(DPSCs)的生物学功能进行对比研究,探讨对骨改建的影响。方法:有限稀释法分离培养SHED和DPSCs,成脂诱导14d进行油红O染色、成骨诱导21d进行茜素红染色;ELISA法检测IL-6、TNF-α分泌水平;两种干细胞常规培养及成骨诱导7d后进行ALP染色、实时定量PCR检测Runx2、RANKL、OPG基因表达。结果:分离获得的SHED和DPSCs具有成骨、成脂分化能力,符合干细胞特点。SHED的炎性细胞因子IL-6、TNF-α的表达均高于DPSCs(P<0.01);成骨诱导7d后SHED的ALP活性强于DPSCs(P<0.05);且经成骨诱导后两种干细胞均表达Runx2、OPG和RANKL,但SHED的表达水平明显高于DPSCs(P<0.05)。结论:SHED与DPSCs相比不仅具有较强的成骨分化能力,而且兼具较强的破骨能力,提示SHED可能参与骨改建调控机制。

Objective： Research differences of biological functions between stem cells from human exfoliated deciduous teeth and dental pulp stem cells from different angles,to explore the influence of bone reconstruction. Methods： SHED and DPSCs were isolated, purified and cultured in vitro by limiting diluction, and oil red O staining at 14 days of adipogenic induction,alizarin red staining at 21 days of osteogenic induction; detect IL-6 and TNF-o~ secretion level by Elisa method; ALP staining at 7 days of two kinds of stem cells in conventional cultivation and osteogenic induced, and detect Runx2, RANKL and OPG gene expression by real-time PCR. Results： Both of SHED and DPSCs have abilty to adipogenic and osteogenic differentiation; conforms to the characteristics of the stem cells. SHED expression of IL-6 and TNF-ct were higher than that of DPSCs（P〈 0.05）; Both kinds of stem cells express Runx20PG and RANKL after osteoinductive differentiation, but the expression of SHED obviously higher than that of DPSCs（P〈0.05）. Conclusion： Compared with DPSCs, SHED has not only the ability of osteogenic differentiation but also an osteoclast capacity,remind that SHED may participate in the bone reconstruction.