联合应用纳米金和磷酸钙骨水泥支架促进牙髓干细胞的增殖和成骨分化

Calcium phosphate cement scaffold with gold nanoparticleson promotes the proliferation and osteogenic differentiation of dental pulp stem cells

目的:磷酸钙骨水泥(calcium phosphate cement,CPC)支架以其良好的生物相容性和骨引导活性而广泛应用于组织工程骨缺损的修复中,但是其骨引导活性有限。因此考虑联合应用纳米金(gold nanoparticles,Au NPs)以促进牙髓干细胞的增殖和成骨分化。方法:以CPC支架作为基底接种牙髓干细胞,在培养基中添加5μg/m L Au NPs进行干细胞培养,以常规培养基组为对照。用透射电镜检测Au NPs的形貌。以荧光显微镜检测细胞接种4 h后的黏附状态,以CCK-8检测细胞增殖情况,检测细胞碱性磷酸酶的活性,并进行茜素红染色以说明矿化物形成情况。结果:Au NPs为均匀的球形,直径20 nm左右。细胞接种后4 h,实验组和对照组的细胞黏附状态没有显著差异。CPC上Au NPs培养基中的细胞14 d时数量多于对照组(P<0.05);且Au NPs培养基中细胞的碱性磷酸酶活性显著增高(P<0.05),矿化物的形成也显著增多(P<0.05)。结论:CPC支架联合Au NPs可显著促进牙髓干细胞的增殖和成骨分化。

Objective：Calcium phosphate cement（CPC）scaffold is widely used in bone tissue engineering due to its goodbiocompatibility and osteoconduction properties. However,it needs further improvements to widen its application. In this study,CPCscaffold with gold nanoparticles（Au NPs）were used to promote the proliferation and osteogenic differentiation of dental pulp stem cells（DPSCs）. Methods：The morphology of Au NPs was detected by transmission electron microscope（TEM）. The cells were seeded onCPC,and cultured in media containing 5 μg/m L Au NPs. Those seeded on CPC and cultured in normal media was set as control. Thecell adhesion was detected by fluorescent stain 4 h after seeding. The cell proliferation was detected by CCK-8,and the alkalinephosphatase（ALP）activity was measured. Alizarin red staining（ARS）was used to detect the mineral synthesis. Results：TEM imageshowed that Au NPs were spherical with 20 nm in average diameter. Fluorescent images showed no significant difference in celladhesion between the two groups. Significant difference in cell proliferation was detected at 14 days（P〈0.05）. ALP activity of theAu NPs media group was significantly increased（P〈0.05）,and the mineralization formation of DPSCs was also increased comparedwith that of cells in normal media（P〈0.05）. Conclusion：It was concluded that CPC scaffold with Au NPs can promote theproliferation and osteogenic differentiation of DPSCs.