Synergistically enhanced osteogenic differentiation of human mesenchymal stem cells by culture on nanostructured surfaces with induction media.
Synergistically enhanced osteogenic differentiation of human mesenchymal stem cells by culture on nanostructured surfaces with induction media.
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DOI：
10.1021/bm100374n
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发表时间：
2010-07-12
影响因子：
6.2
通讯作者：
Suh KY
中科院分区：
文献类型：
--
作者：
You MH;Kwak MK;Kim DH;Kim K;Levchenko A;Kim DY;Suh KY
我们检测了表面纳米形貌对人骨髓间充质干细胞(HMSCs)体外成骨的影响。利用紫外光辅助毛细管力光刻技术制备了具有点(150、400、600 nm直径)和线条(150、400、600 nm宽)的可伸缩(4 cm×5 cm)、清晰的紫外光固化聚氨酯聚合物纳米结构基板。用碱性磷酸酶(ALP)、RT-PCR和实时荧光定量聚合酶链式反应(Real-time PCR)检测第8天对hMSCs成骨分化的影响。我们发现，在成骨诱导介质中，培养在纳米结构表面的hMSCs表现出明显高于未形成图案的PUA表面(对照组)的碱性磷酸酶活性。特别是，400 nm斑点图案上的hMSCs显示出最高水平的碱性磷酸酶活性。进一步的实时定量RT-PCR分析表明，在成骨诱导培养的400 nm斑点图案上培养的hMSCs中，核心结合因子1(Cbfa1)、骨桥蛋白(OP)和骨钙素(OC)的表达水平显著升高。这些结果表明，表面纳米造影术可以协同生物化学诱导物质促进成骨分化。
We have examined the effects of surface nanotopography on in vitro osteogenesis of human mesenchymal stem cells (hMSCs). UV-assisted capillary force lithography was employed to fabricate a scalable (4 cm × 5 cm), well-defined nanostructured substrate of a UV curable polyurethane polymer with dots (150-, 400-, 600-nm diameter) and lines (150-, 400-, 600-nm width). The influence of osteogenic differentiation of hMSCs was characterized at day 8 by alkaline phosphatase (ALP) assay, RT-PCR, and real time PCR analysis. We found that hMSCs cultured on the nanostructured surfaces in osteogenic induction media showed significantly higher ALP activity compared to unpatterned PUA surface (control group). In particular, the hMSCs on the 400-nm dot pattern showed the highest level of ALP activity. Further investigation with real-time quantitative RT-PCR analysis demonstrated significantly higher expression of core binding factor 1 (Cbfa1), osteopontin (OP), and osteocalcin (OC) levels in hMSCs cultured on the 400-nm dot pattern in osteogenic induction media. These findings suggest that surface nanotopography can enhance osteogenic differentiation synergistically with biochemical induction substance.
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