The osteogenic differentiation of human bone marrow MSCs on HUVEC-derived ECM and β-TCP scaffold.
The osteogenic differentiation of human bone marrow MSCs on HUVEC-derived ECM and β-TCP scaffold.
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DOI：
10.1016/j.biomaterials.2012.06.061
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发表时间：
2012-10
期刊：
影响因子：
14
通讯作者：
Yang, Yunzhi
中科院分区：
文献类型：
--
作者：
Kang, Yunqing;Kim, Sungvvoo;Bishop, Julius;Khademhosseini, Ali;Yang, Yunzhi
细胞外基质 (ECM) 在细胞迁移、附着和细胞发育中发挥着关键作用。在这里，我们报道源自人脐静脉内皮细胞（HUVEC）的ECM促进人骨髓间充质干细胞（hMSC）的成骨分化。我们首先通过 HUVEC 接种、孵育和去细胞化在三维 (3D) β-磷酸三钙 (β-TCP) 支架上生产 HUVEC 衍生的 ECM。然后通过 SEM、FTIR、XPS 和免疫荧光染色对 HUVEC 衍生的 ECM 进行表征。随后检查了含有 β-TCP 支架的 HUVEC 衍生 ECM 对 hMSC 成骨分化的影响。 SEM 图像表明，支柱和孔壁表面沉积有致密的基质层。 FTIR 和 XPS 测量显示，与单独的 β-TCP 支架相比，ECM/β-TCP 支架中存在新的官能团（酰胺和羟基）和元素（C 和 N）。免疫荧光图像表明支架上存在高水平的纤连蛋白和 IV 型胶原以及低水平的层粘连蛋白。与单独的 β-TCP 支架相比，含有 ECM 的 β-TCP 支架显着增加了 hMSC 中碱性磷酸酶 (ALP) 的比活性，并上调了成骨相关基因（如 runx2、碱性磷酸酶、骨桥蛋白和骨钙蛋白）的表达。这种增加的效果是由于 MAPK/ERK 信号通路的激活，因为使用 ERK 抑制剂 PD98059 破坏该通路会导致这些成骨基因的下调。含有磷酸钙支架的细胞源性 ECM 是一种有前景的骨组织再生中促进成骨的骨空隙填充剂。关键词：细胞外基质；脐静脉内皮细胞；间充质干细胞；成骨； β-TCP
Extracellular matrix (ECM) serves a key role in cell migration, attachment, and cell development. Here we report that ECM derived from human umbilical vein endothelial cells (HUVEC) promoted osteogenic differentiation of human bone marrow mesenchymal stem cells (hMSC). We first produced an HUVEC-derived ECM on a three-dimensional (3D) beta-tricalcium phosphate (β-TCP) scaffold by HUVEC seeding, incubation, and decellularization. The HUVEC-derived ECM was then characterized by SEM, FTIR, XPS, and immunofluorescence staining. The effect of HUVEC-derived ECM containing β-TCP scaffold on hMSC osteogenic differentiation was subsequently examined. SEM images indicate a dense matrix layer deposited on the surface of struts and pore walls. FTIR and XPS measurements show the presence of new functional groups (amide and hydroxyl groups) and elements (C and N) in the ECM/β-TCP scaffold when compared to the β-TCP scaffold alone. Immunofluorescence images indicate that high levels of fibronectin and collagen IV and low level of laminin were present on the scaffold. ECM-containing β-TCP scaffolds significantly increased alkaline phosphatase (ALP) specific activity and up-regulated expression of osteogenesis-related genes such as runx2, alkaline phosphatase, osteopontin and osteocalcin in hMSC, compared to β-TCP scaffolds alone. This increased effect was due to the activation of MAPK/ERK signaling pathway since disruption of this pathway using an ERK inhibitor PD98059 results in down-regulation of these osteogenic genes. Cell-derived ECM containing calcium phosphate scaffolds is a promising osteogenic-promoting bone void filler in bone tissue regeneration. Keywords: Extracellular matrix; HUVEC; hMSC; Osteogenesis; β-TCP
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DOI：
10.1016/j.bbrc.2006.06.110
发表时间：
2006-08-18
影响因子：
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影响因子：
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