RGD-functionalized bioengineered spider dragline silk biomaterial
RGD-functionalized bioengineered spider dragline silk biomaterial
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DOI：
10.1021/bm0607877
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发表时间：
2006-11-13
影响因子：
6.2
通讯作者：
Kaplan, David L.
中科院分区：
文献类型：
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作者：
Bini, Elisabetta;Foo, Cheryl Wong Po;Kaplan, David L.
蜘蛛丝纤维具有显著的机械性能，这表明其组成蛋白可以作为有用的生物聚合物，用于制造组织形成的生物材料支架。克隆并表达了两个包含RGD细胞结合结构域的生物工程蛋白变体，这些变体来自于Nephila clavipes拖丝主要成分的一致序列。用CD和FTIR对工程蚕丝进行了表征，发现经甲醇处理后，工程蚕丝的结构从随机线圈转变为不溶的β -薄片。重组蛋白被加工成薄膜和纤维，并成功地作为生物材料基质用于培养人骨髓基质细胞，在添加成骨刺激剂的情况下诱导其向骨样组织分化。与组织培养塑料相比，重组蛛丝和编码RGD的重组蛛丝均支持增强人骨髓间充质干细胞（hMSCs）向成骨结果的分化。与含有RGD的重组蛛丝蛋白相比，没有RGD的重组蛛丝蛋白显示出增强的骨相关结果，通过钙沉积来测量。基于与我们之前对蚕丝和RGD修饰的研究的比较，当前的结果说明了生物工程蜘蛛丝蛋白进入新的生物材料基质的潜力，同时也强调了蚕丝来源和RGD向细胞呈现模式的细微差异在组织特异性结果方面的重要性。
Spider silk fibers have remarkable mechanical properties that suggest the component proteins could be useful biopolymers for fabricating biomaterial scaffolds for tissue formation. Two bioengineered protein variants from the consensus sequence of the major component of dragline silk from Nephila clavipes were cloned and expressed to include RGD cell-binding domains. The engineered silks were characterized by CD and FTIR and showed structural transitions from random coil to insoluble, beta-sheet upon treatment with methanol. The recombinant proteins were processed into films and fibers and successfully used as biomaterial matrixes to culture human bone marrow stromal cells induced to differentiate into bone-like tissue upon addition of osteogenic stimulants. The recombinant spider silk and the recombinant spider silk with RGD encoded into the protein both supported enhanced the differentiation of human bone marrow derived mesenchymal stem cells ( hMSCs) to osteogenic outcomes when compared to tissue culture plastic. The recombinant spider silk protein without the RGD displayed enhanced bone related outcomes, measured by calcium deposition, when compared to the same protein with RGD. Based on comparisons to our prior studies with silkworm silks and RGD modifications, the current results illustrate the potential to bioengineer spider silk proteins into new biomaterial matrixes, while also highlighting the importance of subtle differences in silk sources and modes of presentation of RGD to cells in terms of tissue-specific outcomes.