Proliferation and odontogenic differentiation of human umbilical cord mesenchymal stem cells and human dental pulp cells co-cultured in hydrogel
Proliferation and odontogenic differentiation of human umbilical cord mesenchymal stem cells and human dental pulp cells co-cultured in hydrogel
复制标题
人脐带间充质干细胞与人牙髓细胞在水凝胶中共培养的增殖和成牙分化
DOI：
10.1016/j.archoralbio.2019.104582
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发表时间：
2020-01-01
影响因子：
3
通讯作者：
Wu, Yu
中科院分区：
文献类型：
--
作者：
Huang, Chunyang;Bao, Lirong;Wu, Yu
目的：本研究旨在探讨人牙髓细胞（human dental pulp cells，hDPC）和人脐带间充质干细胞（human umbilical cord mesenchymal stem cells，hUCMSCs）在骨形态发生蛋白-2（bone morphogenetic protein-2，BMP-2）和水凝胶辅助下建立的三维共培养体系中的增殖和牙向分化情况。将hDPC和hUCMSC在不同浓度的水凝胶中培养，以探索更适合后续实验的浓度。将BMP-2诱导的hUCMSCs和hDPC在水凝胶中共培养。MTT法检测细胞活力。检测牙本质唾液磷蛋白（DSPP）、牙本质基质蛋白-1（DSPP-1）、碱性磷酸酶（ALP）和骨钙素（Osteocalcin）mRNA的表达。结果：hUCMSCs和hDPC可在水凝胶支架中生长增殖。细胞在低浓度水凝胶中的生长速度高于高浓度水凝胶（P < 0.05）。研究表明，0.25%的水凝胶支架比其他组更适合后续实验。与hUCMSCs单培养组和hDPCs单培养组相比，共培养组的细胞增殖能力、碱性磷酸酶活性和矿化结节形成率均显著提高（P < 0.05）。结论：0.25%水凝胶是共培养体系中适宜的水凝胶浓度。共培养组的成牙本质细胞分化能力和矿化能力均强于单培养组，说明共培养条件在一定范围内可以调控细胞的增殖和分化。
Objective: The aim of this study was to evaluate the proliferation and odontogenic differentiation of human dental pulp cells (hDPCs) and human umbilical cord mesenchymal stem cells (hUCMSCs) in three-dimensional co-culture system which was established with the help of bone morphogenetic protein-2 (BMP-2) and hydrogel.Methods: hDPCs and hUCMSCs were cultured in different concentrations of hydrogel to explore the more suitable concentrations for subsequent experiments. hUCMSCs and hDPCs induced by BMP-2 were co-cultured in the hydrogel. MIT assay was used to measure the cell viability. The differentiation into odontoblast-like cells were measured by the mRNA expression of dentin salivary phosphoprotein (DSPP), dentin matrix protein-1 (DMP-1), alkaline phosphatase and osteocalcin. Alizarin red staining was performed for the formation of mineralized nodules.Results: hUCMSCs and hDPCs could grow and proliferate in hydrogel scaffold. The growth rate of cells in lower concentrations hydrogels were higher than that of high concentrations hydrogels (P < 0.05). The study showed that 0.25% hydrogel scaffold was more suitable for subsequent experiments than other groups. Compared with hUCMSCs-monoculture and hDPCs-monoculture, the co-culture groups exhibited more proliferative potential, alkaline phosphatase activity and mineralization nodule formation (P < 0.05). The mRNA expression in co-culture groups were higher than that of hUCMSCs-monoculture, closed to or even higher than that of hDPCs-monoculture.Conclusion: 0.25% hydrogel was the suitable concentration in co-culture system for subsequent experiments. The co-culture groups had stronger abilities of odontoblastic differentiation and mineralization than cells-monoculture groups, indicated that the co-culture conditions could regulate cell proliferation and differentiation within a certain range.