The use of carbon nanotubes to induce osteogenic differentiation of human adipose-derived MSCs in vitro and ectopic bone formation in vivo
The use of carbon nanotubes to induce osteogenic differentiation of human adipose-derived MSCs in vitro and ectopic bone formation in vivo
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DOI：
10.1016/j.biomaterials.2012.03.045
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发表时间：
2012-06-01
期刊：
影响因子：
14
通讯作者：
Watari, Fumio
中科院分区：
文献类型：
--
作者：
Li, Xiaoming;Liu, Haifeng;Watari, Fumio
碳纳米管（CNT）是最受关注的纳米材料之一，具有独特的电学、机械和表面特性，已被证明适合生物医学应用。在本研究中，我们评估了在相同尺寸的多壁碳纳米管（MWNT）和石墨（GP）压块上体外培养的人类脂肪干细胞的附着、增殖、成骨基因表达、ALP/DNA、蛋白质/DNA和矿化。此外，我们还评估了这两种压块对体内异位骨形成的影响。首先，与 GP 压块相比，MWNT 压块具有更高的吸附蛋白质的能力。在常规培养过程中，结果表明MWNTs可以诱导ALP、cbfa1和COLIA1基因的表达，而GP则不能。此外，MWNTs压块上的细胞的碱性磷酸酶(ALP)/DNA和蛋白质/DNA显着高于GP压块上的细胞。预先在含有50%胎牛血清（FBS）的培养基中吸附蛋白质，发现MWNT压块的ALP/DNA和蛋白质/DNA的增量分别显着大于GP压块的增量，这表明MWNT上吸附更多的蛋白质是至关重要的。更多结果表明，预浸泡在添加了rhBMP-2的培养基中的两种压块上的细胞的ALP/DNA和蛋白质/DNA均高于重新浸泡在添加有50%FBS的培养基中的样品上的细胞，并且MWNTs压块的这些值增加得更多。第 7 天时，MWNT 压块上的矿物质含量比 GP 压块上的矿物质含量更高。体内实验表明，MWNT 可以诱导 ddy 小鼠背侧肌肉组织异位骨形成，而 GP 则不能。结果表明，多壁碳纳米管可能通过浓缩更多蛋白质（包括骨诱导蛋白）来刺激软组织中的诱导细胞形成诱导骨。 (c) 2012 Elsevier Ltd. 保留所有权利。
Carbon nanotubes (CNTs), one of the most concerned nanomaterials, with unique electrical, mechanical and surface properties, have been shown suitable for biomedical application. In this study, we evaluated attachment, proliferation, osteogenic gene expression, ALP/DNA, protein/DNA and mineralization of human adipose-derived stem cells cultured in vitro on multi-walled carbon nanotubes (MWNTs) and graphite (GP) compacts with the same dimension. Moreover, we assessed the effect of these two kinds of compacts on ectopic bone formation in vivo. First of all, higher ability of the MWNTs compacts to adsorb proteins, comparing with the GP compacts, was shown. During the conventional culture, it was shown that MWNTs could induce the expression of ALP, cbfa1 and COLIA1 genes while GP could not. Furthermore, alkaline phosphatase (ALP)/DNA and protein/DNA of the cell on the MWNTs compacts, was significantly higher than those of the cells on the GP compacts. With the adsorption of the proteins in culture medium with 50% fetal bovine serum (FBS) in advance, the increments of the ALP/DNA and protein/DNA for the MWNTs compacts were found respectively significantly more than the increments of those for the GP compacts, suggesting that the larger amount of protein adsorbed on the MWNTs was crucial. More results showed that ALP/DNA and protein/DNA of the cells on the two kinds of compacts pre-soaked in culture medium having additional rhBMP-2 were both higher than those of the cells on the samples re-soaked in culture medium with 50% FBS, and that those values for the MWNTs compacts increased much more. Larger mineral content was found on the MWNTs compacts than on the GP compacts at day 7. In vivo experiment showed that the MWNTs could induce ectopic bone formation in the dorsal musculature of ddy mice while GP could not. The results indicated that MWNTs might stimulate inducible cells in soft tissues to form inductive bone by concentrating more proteins, including bone-inducing proteins. (c) 2012 Elsevier Ltd. All rights reserved.