Chemically-conjugated bone morphogenetic protein-2 on three-dimensional polycaprolactone scaffolds stimulates osteogenic activity in bone marrow stromal cells.
Chemically-conjugated bone morphogenetic protein-2 on three-dimensional polycaprolactone scaffolds stimulates osteogenic activity in bone marrow stromal cells.
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DOI：
10.1089/ten.tea.2010.0132
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发表时间：
2010-07
影响因子：
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通讯作者：
Huina Zhang;Francesco Migneco;Chia-Ying Lin;S. Hollister
中科院分区：
文献类型：
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作者：
Huina Zhang;Francesco Migneco;Chia-Ying Lin;S. Hollister
聚ε-己内酯（PCL）在骨组织工程中受到广泛关注。但PCL的成骨诱导能力不足，限制了其应用。本研究的目的是通过交联偶联法将骨形态发生蛋白-2 （BMP-2）直接附着在PCL支架上，并研究结合后的BMP-2在体外是否保持生物活性。对BMP-2的免疫荧光染色和定量酶联免疫吸附测定表明，BMP-2通过氨解和随后的化学偶联成功地固定在PCL三维支架上。与物理吸附相比，偶联的固定化效率更高。在15天内，偶联BMP-2在PCL支架中的释放速度明显慢于吸附BMP-2的PCL支架，这导致更多的BMP-2局部保留在偶联支架上。此外，在bmp -2结合的PCL支架上培养的骨髓基质细胞中，下游Smads通路上调。最后，在BMP-2偶联的PCL支架上培养的骨髓基质细胞中，成骨标志物（碱性磷酸酶、破骨素和I型胶原）的基因表达上调，而在BMP-2吸附后的PCL支架上则没有上调。因此，我们的发现表明BMP-2在聚酯支架上的偶联是赋予支架骨诱导能力的一种可行的方法。
Poly(ε-caprolactone) (PCL) has received considerable attention in bone tissue engineering. However, the lack of osteoinductive ability of PCL limits its application. The aim of this study was to directly attach bone morphogenetic protein-2 (BMP-2) to PCL scaffolds by a crosslinking conjugation method and to investigate whether the bound BMP-2 maintained bioactivity in vitro. Immunofluorescent staining against BMP-2 and quantitative enzyme-linked immunosorbent assay measurements demonstrated that BMP-2 was successfully immobilized on the PCL three-dimensional scaffold by aminolysis and subsequent chemical conjugation. Conjugation produced much higher immobilization efficiency than the physical adsorption. Conjugated BMP-2 release from the PCL scaffolds was significantly slower than that from BMP-2-adsorbed PCL scaffolds over 15 days, which resulted in more BMP-2 locally retained on the conjugated scaffold. Further, the downstream Smads pathway was upregulated in bone marrow stromal cells cultured on the BMP-2-conjugated PCL scaffolds. Finally, gene expression of osteogenic markers (alkaline phosphatase, osteoclacin, and type I collagen) was upregulated in bone marrow stromal cells cultured on the PCL scaffolds with BMP-2 conjugation, but not on PCL scaffolds after BMP-2 adsorption. Therefore, our finding demonstrated that BMP-2 conjugation on polyester scaffolds is a feasible way to impart scaffolds with osteoinductive capability.