Comparison of osteogenic differentiation capacity in mesenchymal stem cells derived from human amniotic membrane (AM), umbilical cord (UC), chorionic membrane (CM), and decidua (DC)
Comparison of osteogenic differentiation capacity in mesenchymal stem cells derived from human amniotic membrane (AM), umbilical cord (UC), chorionic membrane (CM), and decidua (DC)
复制标题
人羊膜（AM）、脐带（UC）、绒毛膜（CM）和蜕膜（DC）来源的间充质干细胞的成骨分化能力比较
DOI：
10.1186/s13578-019-0281-3
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发表时间：
2019-02-11
影响因子：
7.5
通讯作者：
Zhao, Hai
中科院分区：
文献类型：
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作者：
Shen, Chongyang;Yang, Chuan;Zhao, Hai
骨髓间充质干细胞(MSCs)因其具有成骨分化能力，在骨组织工程领域作为一种有前途的治疗剂已被广泛研究。本研究比较了四种胎盘来源的MSCs的成骨分化能力以及纤维连接蛋白和层粘连蛋白对成骨分化的影响，以确定骨重建组织工程的理想来源。结果茜素红染色检测了四种类型的MSCs的成骨分化水平，并对其钙水平和碱性磷酸酶(ALP)活性进行了定量。此外，本研究还利用免疫印迹法检测了成骨细胞分化蛋白标记物Osterix、I型胶原、骨桥蛋白和骨钙素。用定量聚合酶链式反应和EDU标记分析成骨分化的动力学。测定钙沉积水平、碱性磷酸酶(ALP)活性、骨桥蛋白和骨钙素浓度，以证实细胞外基质(ECM)成分在MSCs成骨分化中的作用。结果表明，从胎盘不同层面分离的MSCs具有不同的成骨分化潜能。重要的是，AM-MSCs和UC-MSCs比CM-MSCs和DC-MSCs更有效和更快地分化为成骨细胞，这与其增殖能力下降有关。在不同类型的MSCs中，AM-MSCs和UC-MSCs由于Akt和ERK途径中的磷酸化增强而具有较高的纤维连接蛋白诱导成骨分化潜能。结论AM-MSCs和UC-MSCs具有较高的成骨能力，纤维连接蛋白能显著增强Akt和ERK途径的成骨能力。
BackgroundMesenchymal stem cells (MSCs) have been extensively explored as a promising therapeutic agent in the field of bone tissue engineering due to their osteogenic differentiation ability. In this study, the osteogenic differential ability and the effect of fibronectin and laminin on the osteogenic differentiation in four types of MSCs derived from placental tissue are compared to determine the ideal source for bone reconstruction tissue engineering.ResultsThe present study examines the osteogenic differentiation levels of four types of MSCs using alizarin red staining and quantifies the calcium levels and alkaline phosphatase (ALP) activity. In addition, this study examines the osteoblast differentiation protein markers osterix, collagen I, osteopontin, and osteocalcin using a Western blot assay. qPCR and EdU labeling assays were employed to identify the kinetics of osteogenic differentiation. Calcium deposit levels, ALP activity, and osteopontin and osteocalcin concentrations were determined to confirm the role of Extracellular matrix (ECM) components role on the osteogenic differentiation of MSCs. The data demonstrated that MSCs isolated from different layers of placenta had different potentials to differentiate into osteogenic cells. Importantly, AM-MSCs and UC-MSCs differentiated into the osteoblast stage more efficiently and quickly than CM-MSCs and DC-MSCs, which was associated with a decrease in their proliferation ability. Among the different types of MSCs, AM-MSCs and UC-MSCs had a higher osteogenic differentiation potential induced by fibronectin due to enhanced phosphorylation during the Akt and ERK pathways.ConclusionsTaken together, these results indicate that AM-MSCs and UC-MSCs possess a higher osteogenic potential, and fibronectin can robustly enhance the osteogenic potentialofthe Akt and ERK pathways.