1,25(OH)_2维生素D_3矿化液对人牙髓干细胞成骨方向诱导作用的研究

Effect of 1,25(OH)_2 vitamin D_3 medium on inducing osteo-differentiation of dental pulp stem cells

目的研究维生素D3矿化液对人牙髓干细胞的成骨方向诱导是否有促进作用。方法从恒牙牙髓中分离培养成纤维样细胞并测试其间充质干细胞的特异性标记物Stro-1阳性率,用一定浓度的1,25(OH)2维生素D3、抗坏血酸、β-甘油磷酸钠的矿化液诱导牙髓干细胞,通过倒置相差显微镜、碱性磷酸酶染色、Vonkossa染色及骨钙素、Ⅰ型胶原基因表达观察和检测矿化液诱导后细胞形态变化及矿化基质分泌情况,以未诱导组为对照。结果维生素D3矿化液连续诱导21d后,诱导组的细胞碱性磷酸酶染色阳性、Ⅰ型胶原和骨钙素的基因表达阳性,并可见明显钙结节形成。结论维生素D3矿化液可以诱导人牙髓干细胞向成骨样细胞分化并促进其产生矿化基质。

Objective To investigate the effect of 1,25（OH）2 vitamin D3 medium on inducing osteo-differentiation of dental pulp stem cells（DPSCs）.Methods Human dental pulp stem cells were isolated from vigorous colony and cultured with 1,25（OH）2 vitamin D3 medium.After osteoinduction for 7,14,21 days,the cells were fixed and HE stained.Alkaline phosphatase staining（ALP） and Vonkossa staining were observed in theses cells.RNA for OC and collagen I were also measured by RT-PCR.Results After 21 days of culture,the positive rate of ALP and Vonkossa staining in vitamin D3 treated DPSCs was significantly higher than in control groups,and the RNA of OC and collagen I in vitamin D3 treated groups were positive.Conclusion 1,25（OH）2 vitamin D3 medium have the ability to induce DPSCs osteo-differentiation.