Cavendish Laboratory, University of Cambridge, Cambridge CB3 0HE, UK.
Nanotechnology. 2013 Jul 5;24(26):265102. doi: 10.1088/0957-4484/24/26/265102.
We study the spectral characteristics of bovine serum albumin (BSA) protein conjugated single-wall carbon nanotubes (SWNTs), and quantify their uptake by macrophages. The binding of BSA onto the SWNT surface is found to change the protein structure and to increase the doping of the nanotubes. The G-band Raman intensity follows a well-defined power law for SWNT concentrations of up to 33 microg ml(-1) in aqueous solutions. Subsequently, in vitro experiments demonstrate that incubation of BSA-SWNT complexes with macrophages affects neither the cellular growth nor the cellular viability over multiple cell generations. Using wide spot Raman spectroscopy as a fast, non-destructive method for statistical quantification, we observe that macrophages effectively uptake BSA-SWNT complexes, with the average number of nanotubes internalized per cell remaining relatively constant over consecutive cell generations. The number of internalized SWNTs is found to be approximately 30 10(6) SWNTs/cell for a 60 mm(-2) seeding density and approximately 100 x 10(6) SWNTs/cell for a 200 mm(-2) seeding density. Our results show that BSA-functionalized SWNTs are an efficient molecular transport system with low cytotoxicity maintained over multiple cell generations.
我们研究了牛血清白蛋白(BSA)与单壁碳纳米管(SWNTs)结合物的光谱特性,并定量研究了巨噬细胞对其的摄取。研究发现,BSA 结合到 SWNT 表面会改变蛋白质结构并增加纳米管的掺杂。在水溶液中,SWNT 浓度高达 33μg/ml 时,G 带 Raman 强度遵循明确的幂律关系。随后的体外实验表明,BSA-SWNT 复合物与巨噬细胞孵育不会影响细胞生长或多个细胞世代的细胞活力。使用宽光斑 Raman 光谱作为一种快速、非破坏性的统计定量方法,我们观察到巨噬细胞有效地摄取了 BSA-SWNT 复合物,每个细胞内化的纳米管数量在连续的细胞世代中保持相对稳定。对于 60mm(-2)的接种密度,内化的 SWNTs 数量约为每个细胞 30×10(6)根 SWNTs,对于 200mm(-2)的接种密度,内化的 SWNTs 数量约为每个细胞 100×10(6)根 SWNTs。我们的结果表明,BSA 功能化的 SWNTs 是一种有效的分子传输系统,具有较低的细胞毒性,并能在多个细胞世代中保持稳定。