Flow perfusion culture of human mesenchymal stem cells on silicate-substituted tricalcium phosphate scaffolds
Flow perfusion culture of human mesenchymal stem cells on silicate-substituted tricalcium phosphate scaffolds
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DOI：
10.1016/j.biomaterials.2008.03.003
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发表时间：
2008-06-01
期刊：
影响因子：
14
通讯作者：
Mygind, Tina
中科院分区：
文献类型：
--
作者：
Bjerre, Lea;Bunger, Cody E.;Mygind, Tina
自体骨移植目前是治疗大面积骨缺损的金标准，但由于供区发病率，其可用性有限。已经提出了不同的替代品来替代这些移植物，本研究提出了一种骨组织工程替代品，使用硅酸盐取代的磷酸三钙（Si-TCP）支架，接种人骨髓间充质干细胞（hMSC），将细胞接种到支架上，并静态培养或在灌注生物培养基中培养。反应器中培养长达21天，并通过碱性磷酸酶活性测定和骨标记物的定量实时RT-PCR评估成骨分化。在培养过程中，来自流动培养的构建体的细胞表现出改善的增殖和成骨分化，这通过几种骨标志物（例如碱性磷酸酶、骨桥蛋白、Runx 2、骨唾液蛋白II和骨形态发生蛋白2）的更显著表达来验证。在流动培养中，细胞和基质均匀地分布在整个支架中，而静态培养后仅获得外周层。在动态培养中产生了具有上级成骨特性的活的和均质的离体骨结构，并且可以提供自体移植物的替代物。(c)2008爱思唯尔有限公司保留所有权利。
Autologous bone grafts are currently the gold standard for treatment of large bone defects, but their availability is limited due to donor site morbidity. Different substitutes have been suggested to replace these grafts, and this study presents a bone tissue engineered alternative using silicate-substituted tricalcium phosphate (Si-TCP) scaffolds seeded with human bone marrow-derived mesenchymal stem cells (hMSC), The cells were seeded onto the scaffolds and cultured either statically or in a perfusion bio-reactor for up to 21 days and assessed for osteogenic differentiation by alkaline phosphatase activity assays and by quantitative real-time RT-PCR on bone markers. During culture, cells from the flow cultured constructs demonstrated improved proliferation and osteogenic differentiation verified by a more pronounced expression of several bone markers, e.g. alkaline phosphatase, osteopontin, Runx2, bone sialoprotein II, and bone morphogenetic protein 2. Cells and matrix were distributed homogenously throughout the entire scaffold in flow culture, whereas only a peripheral layer was obtained after static culture. A viable and homogenous ex vivo bone construct with superior osteogenic properties was produced in dynamic culture and may provide a replacement for autologous grafts. (c) 2008 Elsevier Ltd. All rights reserved.